UA110317C2 - Process for the preparation of feed materiala - Google Patents

Abstract

The invention relates to a method of preparation of feed material consisting of irradiation of biomass by an electron beam with a radiation dose of at least 5.0 Mrad and a power of at least 5 kW, to obtain feed material having an average molecular weight of from 3000 Daltons to 50,000 Daltons.

Description

TECHNICAL FIELD

This invention relates to the processing of biomass, compositions that include saccharide elements organized into a molecular chain, methods of obtaining amino acids or antibiotics, methods of obtaining food or immunostimulating material and products obtained by such methods.

TECHNICAL LEVEL

Biomass, particularly biomass waste, is widely available. It would be useful to obtain products from biomass.

ESSENCE OF THE INVENTION

Illustrative products that can be produced using the methods presented herein include food products suitable for consumption, for example, in human and/or animal consumption, in aquaculture, agriculture, growing plants without soil (hydroponics), pharmaceuticals, nutraceuticals, pharmaceutical delivery vehicles and dosage forms, pharmaceutical excipients, pharmaceutical conjugates, cross-linked matrices such as hydrogels, absorbent materials, fertilizers and lignin products. Any product described in this document or produced by the methods described in this document may be used as is or as a precursor or intermediate in the preparation of another product.

In many embodiments, products can be manufactured using Maiig! Eogse "M

Spitiwig In the ways of Maishga! Rogse"M Spetivzigu uses the controlled application and manipulation of physical forces, such as particle beams, gravity, light, etc., to introduce predictable structural and chemical changes into molecules. In preferred embodiments, the methods of Maiig! Rogse"M Spetivzigu change the molecular structure without chemical reagents or microorganisms. With the use of natural processes, it is possible to create a new useful material without harming the environment.

In one aspect, producing a food material includes altering the molecular structure of biomass polysaccharides, including polysaccharides in the form of cellulose, hemicellulose or starch, to produce a food material that has nutrient availability greater than that of the biomass.

In one aspect, the present invention relates to methods of preparing food materials for animals (e.g., human and animals, including, but not limited to, beef and dairy cattle, pets, zoo animals, etc.) and for plants (e.g., agricultural plants or crops or aquatic plants, in particular in a hydroponic solution or in aquaculture), and for aquatic organisms (for example, fish, crustaceans, molluscs, etc.).

These methods include obtaining a first material that includes biomass (for example, plant biomass, animal biomass, microbial biomass and municipal waste biomass) containing polysaccharides in the form of cellulose, hemicellulose and/or starch. The polysaccharide molecular structure of the first material is then modulated (eg, increased, decreased, or preserved) to produce a second material with greater availability of nutrients (eg, proteins, carbohydrates, fats, vitamins, and/or minerals) than the first material.

Optionally, the methods may include administering the second material to an animal (eg, a human and/or non-human animal).

In some embodiments, the methods described herein can be used to produce materials suitable for use in supporting or stimulating the growth of microorganisms (e.g., bacteria, yeast, fungi, unicellular organisms, such as algae, or fungi-like protozoa, such as slime molds), aquatic organisms (for example, in aquaculture) and/or plants and trees (for example, in agriculture, growing plants without soil and forestry).

In one aspect, the method includes converting the processed material using a microorganism to produce an edible material, an amino acid or derivative thereof, an antibiotic, or an immunostimulating material, wherein the processed material is obtained by processing biomass containing polysaccharides in the form of cellulose, hemicellulose or starch having a first level refractoriness, using at least one of radiation exposure, sonication, pyrolysis and oxidation, to obtain a processed material having a refractoriness level lower than the refractoriness level of the first material, where refractoriness is determined by incubation in the presence of cellulase.

Some embodiments of obtaining edible material include isolation and/or purification of edible material. Edible material can be digested and/or absorbed. The edible material may be selected from the group consisting of pharmaceuticals, nutraceuticals, proteins, fats, vitamins, oils, fibers, minerals, sugars, carbohydrates, and alcohol.

In some embodiments of the preparation of an amino acid or its derivative, the amino acid or its derivative is selected from the group consisting of I-amino acids and Y-amino acids, such as I-glutamic acid (monosodium glutamate (M5C)), I-aspartic acid, I- phenylalanine, I1-lysine, I-threonine, I-tryptophan, I-valine, I-leucine, I-isoleucine, I-methionine, I-histidine and 1-phenylalanine, I-lysine, O-methionine and I-tryptophan. The microorganism can be selected from the group consisting of lactic acid bacteria (LAB), E. soii, Vasiliye 5ibBiivB ii Soguperasiegicht dishatisit.

In some embodiments, the antibiotic is selected from the group consisting of tetracycline, streptomycin, cyclohexamide, neomycin, cycloserine, erythromycin, kanamycin, lincomycin, nystatin, polymyxin B, and bacitracin. The microorganism can be selected from the group consisting of Zigeriotuseb5 geto5y5, Zigeriotusev dgizeyy5, zZigeriotusev iPodiae, Zigeriotuse5 ogspidasey5, Zperiotusez eguiigteiv5, 5Bigeriotusev5

Kapatuseiisiv, Zigeriotusev, zigeriotusez poshgzei, Vasiyi5 roiutuh and Vasiyi5 Ispepiogtiv.

In some embodiments, the biomass may be selected from the group consisting of paper, paper products, paper waste, wood, pressed wood, wood sawdust, agricultural waste, sewage, silage, grasses, rice husk, cake, cotton, jute, hemp , flax, bamboo, sisal, abaca, straw, cores of corn cobs, corn straw, millet, alfalfa, hay, coconut fibers, sea grass, algae and their mixtures. In some cases, the biomass has internal fibers and is comminuted to such an extent that the internal fibers are substantially exposed, and/or where the biomass has a BET surface area of greater than about 0.25 mg/g and a bulk density of less than about 0, 5 g/cm3. Processing may include exposure to ionizing radiation. The processed material can be subjected to enzymatic hydrolysis.

In one aspect, the absorber comprises a processed biomass material comprising saccharide units arranged in a molecular chain, wherein from about 1 out of every 2 to about 1 out of every 250 saccharide units comprises a carboxylic acid group or an ester or salt thereof.

In some embodiments, the recycled biomass material is treated with silane to render the absorbent lipophilic.

In another aspect, the filter material includes an irradiated cellulosic or lignocellulosic material adapted to retain and filter the flow.

In another aspect, the product includes a converted material obtained by converting a recycled material using a microorganism to produce a converted material, wherein the recycled material is obtained by processing biomass containing polysaccharides in the form of cellulose, hemicellulose or starch having a first level of refractoriness using at least one from radiation exposure, sonication, pyrolysis and oxidation, to obtain a processed material having a level of refractoriness lower than the level of refractoriness of the first material, where refractoriness is determined by incubation in the presence of cellulase.

In another aspect, the present invention relates to methods of improving the pharmaceutical profile of materials. These methods include obtaining a first material comprising biomass (e.g., plant biomass, animal biomass, microbial biomass, and municipal waste biomass) containing polysaccharides in the form of cellulose, hemicellulose, and/or starch, and modulating (e.g., increasing, decreasing, or maintaining ) of the molecular structure of the polysaccharides of the first material to obtain the second material, where one of the results of the methods is that the pharmaceutical profile of the second material is better or improved compared to the pharmaceutical profile of the first material. In some cases, the methods include the use of first materials with little or no pharmaceutical profile before modulating the molecular structure of the first material. The second materials obtained using the methods described in this document are suitable for administration to an animal.

In the next aspect, the invention relates to methods of obtaining a pharmaceutical agent of plant origin. These methods include the processing of material comprising biomass (eg plant biomass, animal biomass, microbial biomass and municipal waste biomass) containing polysaccharides in the form of cellulose, hemicellulose and/or starch containing one or more plant-produced pharmaceuticals, using any one or more of radiation exposure, ultrasound treatment, pyrolysis and oxidation to obtain a pharmaceutical agent of plant origin. In some cases, the pharmaceutical preparation of plant origin can be isolated and/or purified.

In another aspect, the present invention relates to methods of preparing nutraceuticals for human and/or non-human animal consumption. These methods include processing biomass-containing material (e.g., plant biomass, animal biomass, microbial biomass, and municipal waste biomass) containing polysaccharides in the form of cellulose, hemicellulose, and/or starch so that the molecular structure of the material's polysaccharides is altered (e.g., increased or the molecular weight of the material decreased). These methods may optionally also include administration of the resulting materials to humans and non-human animals.

In an alternative aspect, the invention relates to methods of obtaining biological agents and/or pharmaceutical agents. These methods include processing material that includes biomass containing polysaccharides in the form of cellulose, hemicellulose and/or starch to change the molecular structure of the material's polysaccharides. The resulting materials can then be combined with one or more biological agents and/or one or more pharmaceutical agents that can be administered to the subject.

The present invention also relates to methods of obtaining hydrogels. These methods include processing material that includes biomass containing polysaccharides in the form of cellulose, hemicellulose and/or starch, and changing the molecular structure of polysaccharides to produce material that includes cross-linked polymer chains. In addition, the method may include cross-linking polymer chains in the recycled material.

In another aspect, the present invention relates to methods of obtaining an absorbing or adsorbing material. These methods include processing the material, which includes biomass containing polysaccharides in the form of cellulose, hemicellulose and/or starch, and changing the molecular structure of the polysaccharides to produce absorbent material. These absorbent materials can be charged, eg positively or negatively charged, and they can have lipophilic and/or hydrophilic properties. Essentially, the materials can be used as litter or litter for animals and/or absorbent material to bind materials in solution (eg pollutants). In some embodiments, these absorbent materials can be used to bind biological materials in blood or plasma solutions.

In the next aspect, the present invention relates to methods of obtaining fertilizers. These methods

These include processing the material, which includes biomass containing polysaccharides in the form of cellulose, hemicellulose and/or starch, and changing the molecular structure of the polysaccharides to produce a material that has a higher solubility than the original material and that is suitable as a fertilizer.

Each of these methods includes processing the biomass using one or more of (e.g., one, two, three, or four of) size reduction (e.g., mechanical size reduction of individual biomass fragments), radiation exposure, sonication, pyrolysis, and oxidation to modulate the materials . In some embodiments, the method uses a dose of radiation, for example, from 0.1 to 10 Mrad.

In some embodiments, the methods use a radiation dose, for example, from more than 10 to 1000 Mrad.

In some aspects, the present invention also relates to a composition made using any of the methods described herein. For example, the invention relates to a composition that includes saccharide units arranged in a molecular chain, wherein from about 1 out of every 2 to about 1 out of every 250 saccharide units contains a carboxylic acid group or an ester or salt thereof, and the composition is suitable for use as a food material .

In some embodiments, the composition includes a plurality of such chains. In some cases, from about 1 in every 5 to about 1 in every 250 saccharide units of each chain contains a carboxylic acid group or an ester or salt thereof, particularly from about 1 in every 8 to about 1 in every 100 or from about 1 in every 10 up to about 1 in every 50 saccharide units of each chain contains a carboxylic acid group or its ester or salt. Each chain can include from about 10 to about 200 saccharide units. Each chain may include hemicellulose or cellulose, and/or each chain may include saccharide units that include groups selected from the group consisting of nitroso groups, nitro groups, and nitrile groups. Saccharide elements can include 5 or b carbon saccharide elements. The average molecular weight of the composition according to the RES standards is from 1,000 to 1,000,000, in particular, less than 10,000.

By "fit for use as a food material" is meant that the composition is non-toxic, under the conditions of its intended use, to the living organism on which it is fed, and provides some nutritional value to the organism, such as energy and/or nutrients (516).

In some embodiments, the biomass feedstock is pre-treated. In some embodiments, the methods described herein may include pretreatment to reduce one or more sizes of individual biomass fragments. For example, pre-processing may include reducing one or more sizes of individual fragments, such as crushing, slicing, grinding, crushing, or grinding.

In all methods described in this document, pressure can be applied. For example, at least one of the treatments, such as radiation exposure, can be performed on the biomass at a pressure greater than about 2.5 atmospheres (0.25 MPa), such as greater than 5 or atmospheres (0.5 or 1 MPa). 10 Examples of biomass (also called "biomass feedstock" or "feedstock") include cellulosic or lignocellulosic materials such as paper, paper products, paper waste, wood, pressed wood, wood sawdust, agricultural waste, sewage, silage, grasses, rice husk, cake, cotton, jute, hemp, flax, bamboo, sisal, abaca, straw, corn cob kernels, corn straw, millet, alfalfa, hay, coir, cassava and synthetic cellulose and/or mixtures thereof. In some cases, the biomass may include unicellular and/or multicellular organisms. illustrative organisms include, but are not limited to, unicellular organisms (e.g., animals (e.g., protozoa such as flagellates, amoebae, ciliates, and sporophytes)) and plants (e.g., algae such as alveolobionts, chlorarachniophytes, cryptomonads, euglenids, glaucophytes , haplophytes, red algae, straminopile and green algae)), sea grass, plankton (e.g. macroplankton, mesoplankton, microplankton, nanoplankton, picoplankton and femptoplankton), phytoplankton, bacteria (e.g. gram-positive bacteria, gram-negative bacteria and extremophiles), yeasts and / or their mixtures. In some cases, the biomass may include unicellular or multicellular organisms obtained from the ocean, lakes and bodies of water, including salt water and fresh water. In some cases, biomass may include organic waste materials such as livestock waste or animal excrement or human waste or excrement (eg compost and sewage). In some cases, the biomass may include any combination of any of these. Other biomass materials are described in this document. Other biomass materials that include cellulose are described in patents, patent applications and publications, which are incorporated herein by reference. In some cases, biomass can be, for example, in solution, dry and frozen.

If the biomass is or includes microorganisms, these microorganisms typically include carbohydrates, such as cellulose. These microorganisms can be in solution, dry, frozen, in an active and/or inactive state. In some embodiments, these microorganisms may require additional processing prior to exposure to the methods described herein. For example, microorganisms can be in solution and can be extracted from the solution, for example, by centrifugation and/or filtration. Alternatively or additionally, the microorganisms can be subjected to the methods described in this document without these additional stages, for example, the microorganisms can be used in solution. In some cases, biomass may be or may include natural or synthetic material.

Irradiation, for example, can be performed using ionizing radiation such as gamma rays, electron beams, or ultraviolet C radiation having a wavelength of about 100 nm to about 280 nm. Ionizing radiation can include electron beam radiation. For example, radiation may be administered at a total dose of from about 10 Mrad to about 150 Mrad, for example at a dose rate of from about 0.5 to about 10 Mrad/day or from 1 Mrad/s to about 10

Mrad/s. In some embodiments, the implementation of irradiation includes the use of two or more sources of radiation, such as gamma rays and an electron beam.

In some embodiments, the biomass exhibits a first level of refractoriness and the carbohydrate material exhibits a second refractoriness level that is lower than the first refractoriness level. For example, the second level of reluctance may be lower than the first level of reluctance by at least about 10 95 (eg, 20, 30, 40, 50, 60, 70, 80, 90, 99, 100 95 ). In some embodiments, the recalcitrance level can be reduced by 50-90 95.

Biomass can be obtained by crushing biomass (eg biomass fiber sources) to provide fibrous material. For example, crushing can be done using a cutting device with a rotating knife. The fibers of the fibrous material may have, for example, an average ratio of length to diameter (G/O) greater than 5/1. The fibrous material may have, for example, a surface area of VET greater than 0.25 mg/g (eg, 0.3, 0.35, 0.35, 0.4, 0.5, 60 1, 1.5, 2, 3 , 10, 25 me/g or more than 25 mg/g).

In some embodiments, the carbohydrate may include one or more p-1,4 linkages and have a number average molecular weight of about 3,000 to 50,000 daltons.

In some examples, the pretreated biomass material may further include a buffer such as sodium bicarbonate or ammonium chloride, an electrolyte such as potassium chloride or sodium chloride, a growth factor such as biotin, and/or a base pair such as uracil, a surfactant substance, mineral or chelating agent.

An enzyme, such as a cellulolytic enzyme, and/or a swelling agent can be used to assist in reducing the molecular weight of cellulose in any of the methods described herein.

When a microorganism is used, it may be a naturally occurring microorganism or an engineered microorganism (eg, a genetically modified microorganism (GM)). For example, a microorganism can be a bacterium, such as a cellulolytic bacterium, a fungus, such as a yeast, a plant, or a single-celled organism such as an algae, a protozoan or a fungus-like single-celled organism, such as a slime mold, a single-celled organism (e.g., an animal (e.g., a protozoan such as a flagellate , amoebae, ciliates and sporozoites) and plants (e.g. algae such as alveolobionts, chlorarachniophytes, cryptomonads, euglenids, glaucophytes, haplophytes, red algae, straminopyles and green algae), sea grass, plankton (e.g. macroplankton, mesoplankton, microplankton, nanoplankton, picoplankton, and femtoplankton), phytoplankton, and/or mixtures thereof. In some embodiments, the microorganism is a white-red mold. In some cases, the microorganism may include unicellular and/or multicellular organisms, such as organisms from the ocean, lakes, and reservoirs, including salt water and fresh water.When the organisms are compatible, it is possible use their mixtures.

Generally, various microorganisms can produce a number of useful products by acting on materials, converting, bioconverting or fermenting materials.

For example, alcohols, organic acids, hydrocarbons, hydrogen, proteins, carbohydrates, fats/oils/lipids, amino acids, vitamins, or mixtures of any of these materials can be produced by fermentation or other methods.

Examples of products that can be obtained using the methods described herein

From the document, include mono- and polyfunctional C1-Cb-alkyl alcohols, mono- and polyfunctional carboxylic acids, C1-Cb-hydrocarbons and their combinations. Specific examples of suitable alcohols include methanol, ethanol, propanol, isopropanol, butanol, ethylene glycol, propylene glycol, 1,4-butanediol, glycerin, and combinations thereof. Specific examples of suitable carboxylic acids include formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, palmitic acid, stearic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, oleic acid, linolenic acid, glycolic acid. , lactic acid, y-hydroxybutyric acid and their combinations.

Examples of suitable hydrocarbons include methane, ethane, propane, pentane, n-hexane and combinations thereof.

Another aspect of the invention relates to a method that includes converting a low molecular weight sugar, or a material that includes a low molecular weight sugar, in a mixture with biomass, a microorganism and a solvent or solvent system, such as water or a mixture of water and an organic solvent, into any product described herein documents Without being bound by any particular theory, it is believed that the presence of a solid, such as a solid with a high surface area and/or high porosity, can increase reaction rates by increasing the effective concentration of solutes and providing a substrate on which to flow reaction. A detailed description of such conversion is provided in US Patent Application Mo. 12/417840, filed Apr. 2009, the entire content of which is incorporated herein by reference in its entirety.

The term "fibrous material", as used in this description, is a material that includes a plurality of loose, discrete and separate fibers. For example, the fibrous material can be obtained from a source of fibers, which is bleached kraft paper, by crushing, for example, using a cutting device with a rotating knife.

The term "sieve", as used herein, means an element capable of sieving material according to size. Examples of sieves include a plate, a cylinder with holes or the like, or a wire sieve or cloth.

The term "pyrolysis", as used in this description, means the destruction of bonds in the material using thermal energy. Pyrolysis can occur when the material in question is in a vacuum or immersed in a gaseous substance such as an oxidizing gas such as air or oxygen or a reducing gas such as hydrogen.

The oxygen content is determined using elemental analysis by pyrolysis of the sample in a furnace operating at 1300 "C or higher.

For the purposes of this description, carbohydrates are materials that consist entirely of one or more saccharide elements or that include one or more saccharide elements. The saccharide elements can be functionalized in the ring region with one or more functional groups, such as carboxylic acid groups, amino groups, nitro groups, nitroso groups, or nitrile groups, and still be considered carbohydrates.

Carbohydrates can be polymeric (e.g., equal to or greater than 10-mer, 100-mer, 1000-mer, 10000-mer, or 100000-mer), oligomeric (e.g., equal to 4-mer, 5-mer, b-mer, 7 -mer, 8-mer, 9U-mer or 10-mer or exceeding them), trimeric, dimeric or monomeric. When carbohydrates are made of more than one repeating element, all the elements can be the same or different.

Examples of polymeric carbohydrates include cellulose, xylan, pectin, and starch, while examples of dimeric carbohydrates include cellobiose and lactose. Examples of monomeric carbohydrates include glucose and xylose.

Carbohydrates can be part of a supramolecular structure, for example, covalently attached to the structure. Examples of such materials include lignocellulosic materials, such as materials found in wood.

A starch material is a material that is starch or a starch derivative or includes significant amounts of starch or a starch derivative, such as more than about 5 wt. 96 starch or derived starch. For the purposes of this description, starch is a material that includes amylose, amylopectin or their physical and/or chemical mixture, for example a mixture of amylose and pectin, which is 20:80 or 30:70 by weight. 95. For example, rice, corn and their mixtures are starchy materials. Starch derivatives include, for example, maltodextrin, acid-modified starch, basic-modified starch, bleached starch, oxidized starch, acetylated starch, acetylated and oxidized starch, phosphate-modified starch, genetically modified starch, and starch that is resistant to cleavage.

For the purposes of this description, a low molecular weight sugar is a carbohydrate or a derivative thereof,

Which have a formula molecular weight (excluding moisture) of less than about 2000, such as less than about 1800, less than about 1600, less than about 1000, less than about 500, less than about 350, or less than about 250. For example, low molecular sugar can be a monosaccharide, for example glucose or xylose, a disaccharide, for example cellobiose or sucrose, or a trisaccharide.

Swelling agents, as used herein, are materials that cause visible swelling, such as an increase in the volume of cellulosic and/or lignocellulosic materials relative to the unswollen state of 2.5 95 when applied to such materials as a solution , for example, an aqueous solution. Examples include alkalines such as sodium hydroxide, potassium hydroxide, lithium hydroxide and ammonium hydroxides, acidifying agents such as mineral acids (e.g. sulfuric acid, hydrochloric acid and phosphoric acid), salts such as zinc chloride, calcium carbonate, sodium carbonate , benzyltrimethylammonium sulfate, and basic organic amines such as ethylenediamine.

In some embodiments, no chemical reagents, such as swelling agents, are added to the biomass prior to irradiation. For example, in some of these embodiments, no alkalis (such as sodium hydroxide, potassium hydroxide, lithium hydroxide, and ammonium hydroxides), acidifiers (such as mineral acids (such as sulfuric acid, hydrochloric acid, and phosphoric acid) are added prior to irradiation or other processing acid)), salts such as zinc chloride, calcium carbonate, sodium carbonate, benzyltrimethylammonium sulfate, or basic organic amines such as ethylenediamine. In some cases, no additional water is added. For example, biomass before processing may have less than 0.5 wt. 95 added chemical reagents, for example less than 0.4, 0.25, 0.15 or 0/1 wt. 96 added chemical reagents. In some cases, the biomass before irradiation has no more than trace amounts, for example less than 0.05 wt. 95, added chemical reagents. In other cases, the biomass has essentially no added chemical reagents or swelling agents before irradiation. Avoiding the use of such chemical reagents can also extend to processing, for example during the whole time before fermentation or during the whole time.

The term "edible" as used herein means fit for consumption as food.

A "ground material" as used herein is a material that includes individual fibers in which at least about 5,095 individual fibers have a length/diameter ratio (G/0) of at least about 5 and which have an uncompressed bulk density less than about 0.6 g/cm3.

In some embodiments, changing the molecular structure of biomass, as used in this description, means changing the location of chemical bonds, such as the type and number of functional groups, or the conformation of the structure. For example, a change in the molecular structure may include a change in the level of recalcitrance of the material, a change in the supramolecular structure of the material, oxidation of the material, a change in the average molecular weight, a change in the average crystallinity, a change in the surface area, a change in the degree of polymerization, a change in the porosity, a change in the degree of branching, graft copolymerization with other materials, changing the size of the crystal domain or changing the size of the entire domain.

Unless otherwise defined, all technical and scientific terms used in this description have the same meaning as is commonly understood by those skilled in the art to which this invention belongs. Although methods and materials similar or equivalent to the methods or materials described herein may be used in the practice or testing of this invention, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned in this description are incorporated by reference in their entirety. In case of contradiction, you need to be guided by this description. In addition, the materials, methods and examples are illustrative only and are not intended to be limiting.

As used herein, the term "subject" is used throughout the specification to describe an animal that is either human or non-human. The term includes, but is not limited to, birds, reptiles, fish, plants, amphibians and mammals such as humans, other primates, pigs, rodents such as mice and rats, rabbits, guinea pigs, hamsters, as well as cows, horses , cats, dogs, sheep and goats.

The entire content of IMO 2008/073186 is incorporated herein by reference in its entirety.

The complete disclosure of each of the following US patent applications is incorporated herein by reference: prior US application serial numbers 61/049391; 61/049394; 61/049395; 61/049404; 61/049405; 61/049406; 61/049407; 61/049413; 61/049415 and 61/049419, filed April 30, 2008; US Patent Application Serial Nos. 61/073432; 61/073436; 61/073496; 61/073530; 61/073665 and 61/073674, filed on June 18, 2008; US Provisional Application Serial No. 61/106861, filed Oct. 20, 2008; US Patent Application Serial Nos. 61/139324 and 61/139453, both filed Dec. 19, 2008, and US Patent Application Serial No. 12/417707; 12/417720; 12/417840; 12/417699; 12/417731; 12/417900; 12/417880; 12/417723; 12/417786 and 12/417904, all filed Apr. 2009.

Any carbohydrate material described herein may be used in any application or method described in any patent or patent application incorporated herein by reference.

In any of the ways described in this document, radiation can be applied from a device that is in storage.

Other features and advantages of the invention will become apparent from the detailed description and claims presented below.

DESCRIPTION DRAWINGS

In Fig. 1 presents a block diagram illustrating the conversion of biomass into products and by-products.

In Fig. 2 is a block diagram illustrating the conversion of a fiber source into a first and a second fibrous material.

In Fig. 3 shows a cross-section of a cutting device with a rotating knife.

In Fig. 4 is a block diagram illustrating the conversion of a fiber source into a first, second, and third fibrous material.

In Fig. 5 is a block diagram illustrating the compaction of the material.

In Fig. 6 is a perspective view of a pelletizing press.

In Fig. 7A shows compacted fibrous material in the form of granules.

In Fig. 7B shows a cross-section of hollow granules in which the center of the cavity is on the same line as the center of the granule.

In Fig. 7C shows a cross-section of a hollow pellet in which the center of the cavity is offset relative to the center of the pellet.

In Fig. 70 shows a cross-section of a three-lobed pellet.

In Fig. 8 is a block diagram illustrating a processing sequence for processing (516) raw materials.

In Fig. 9 shows a sectional view of a gamma emitter located in a concrete storage.

In Fig. 10 shows an enlarged view of area K from fig. 9.

In Fig. 11 presents a block diagram illustrating the sequence of preliminary processing of raw materials by irradiation with an electron beam.

In Fig. 11A is a schematic representation of biomass ionized and then oxidized or quenched.

In Fig. 118 shows a diagram (side view) of a system for irradiating material with a low bulk density, and in Fig. 11C shows a cross-section of the system along the line 110-116.

Fig. 110 schematically shows a cross-sectional view of a fluidized bed system for irradiation of material with a low bulk density.

In Fig. 11E is a diagram (side view) of another system for irradiating material with a low bulk density.

In Fig. 12 shows a diagram of a system for ultrasound treatment of a technological flow of cellulosic material in a liquid medium.

In Fig. 13 shows a diagram of an ultrasonic treatment device having two transducers connected to one horn.

In Fig. 14 presents a block diagram illustrating a system for pyrolytic pretreatment of raw materials.

In Fig. 15 shows a cross-section (side view) of a pyrolysis chamber.

In Fig. 16 shows a cross-section (side view) of a pyrolysis chamber.

In Fig. 17 shows a cross-section (side view) of a pyrolyzer that includes a heated hair.

In Fig. 18 schematically presents a cross-section (side view) of a pyrolyzer at a point

Curie.

In Fig. 19 schematically presents a cross-section (side view) of a furnace pyrolyzer.

In Fig. 20 schematically presents a cross-section (top view) of a laser device for pyrolysis.

From Fig. 21 is a schematic cross-section (top view) of a tungsten hair flash pyrolysis device.

In Fig. 22 is a block diagram illustrating a system for oxidative pretreatment of raw materials.

In Fig. 23 shows a block diagram illustrating the general view of the conversion process

C5 sources of fiber into the product, such as ethanol.

In Fig. 24 shows a cross-sectional view of a steam explosion device.

In Fig. 25 schematically presents a cross-sectional view of a hybrid device for electron beam/ultrasound treatment.

In Fig. 26 presents a picture, obtained with the help of a scanning electron microscope at a magnification of 25X, of a fibrous material obtained from paper with a multilayer coating. The fibrous material was obtained on a rotary knife cutter using a screen with 1/8 inch (0.32 cm) openings.

In Fig. 27 presents a picture obtained with the help of a scanning electron microscope at a magnification of 25X of a fibrous material obtained from bleached kraft cardboard. The fibrous material was obtained on a rotary knife cutter using a screen with 1/8 inch (0.32 cm) openings.

In Fig. 28 presents a picture obtained with the help of a scanning electron microscope at a magnification of 25X of a fibrous material obtained from bleached kraft cardboard. The fibrous material was crushed twice on a rotary knife cutter using a screen with 1/16 inch (0.16 cm) openings for each crushing.

In Fig. 29 presents a picture obtained with the help of a scanning electron microscope at a magnification of 25X of a fibrous material obtained from bleached kraft cardboard. The fibrous material was crushed three times on a cutting device with a rotating knife. A 1/8 inch (0.32 cm) sieve was used during the first crushing; a 1/16 inch (0.16 cm) sieve was used for the second crushing and a 1/32 inch (0.08 cm) sieve was used for the third crushing.

In Fig. 30 is a schematic side view of the device for ultrasound treatment, and in Fig. 31 is a cross-sectional view of the processing cell of FIG. 30.

In Fig. 32 is a scanning electron microscope image, taken at 1000X magnification, of fibrous material obtained by crushing millet on a rotary knife cutter and then passing the crushed material through a 1/32 inch (0.08 cm) screen.

In Fig. 33 and 34 show images obtained using a scanning electron microscope at a magnification of 1000X of the fibrous material from Fig. 32 after irradiation with gamma rays at a dose of 10 Mrad and 100 Mrad, respectively.

In Fig. 35 presents images obtained using a scanning electron microscope at a magnification of 1000X of the fibrous material from Fig. 32 after irradiation with a dose of 10 Mrad and treatment with ultrasound.

In Fig. 36 presents images obtained using a scanning electron microscope at a magnification of 1000X of the fibrous material from Fig. 32 after irradiation with a dose of 100 Mrad and treatment with ultrasound.

In Fig. 37 presents the infrared spectrum of kraft paper cut on a cutting device with a rotating knife.

In Fig. 38 presents the infrared spectrum of the kraft paper from Fig. 37 after exposure to gamma radiation at a dose of 100 Mrad.

In Fig. 39 shows a diagram of the biomass conversion process.

In Fig. 40 shows a diagram of another biomass conversion process.

In Fig. 41 presents a diagram of a mobile biomass processing plant based on a truck.

In Fig. 42 shows a diagram of a mobile plant for processing biomass on the basis of a train.

In Fig. 43A and 43B are diagrams showing processing stages for obtaining products and co-products from biomass (A) and for obtaining products using the bioconversion stage.

In Fig. 44 is a diagram showing the fermentation process with variable volume feeding.

In Fig. 45 is a diagram showing the fermentation process with fixed volume feeding.

In Fig. 46 presents a diagram showing the stages of processing necessary to obtain products 1, 2 and 3. The asterisks indicate that the stage is optional. The black arrow indicates that

From which it is possible to carry out an optional compaction stage.

DETAILED DESCRIPTION

Biomass (eg, plant biomass, animal biomass, microbial biomass, and municipal waste biomass) can be processed into useful products using the methods described herein, such as food products. In addition, functionalized materials having the desired types and amounts of functional groups, such as carboxylic acid groups, aldehyde groups, ketone groups, nitrile groups, nitro groups, or nitroso groups, can be obtained using the methods described herein.

Such functionalized materials can be, for example, more soluble, more easily utilized by various microorganisms, or they can be more stable over a long time, for example, less prone to oxidation. In this specification, below, systems and processes are described in which various biomass materials can be used, such as cellulosic materials, lignocellulosic materials, starch materials, or materials that are or include low molecular weight sugars as raw materials. Biomass materials are often readily available, but may be difficult to process, for example by fermentation, or they may give suboptimal yields at sufficiently low rates, for example by fermentation. Biomass materials are first pre-treated, often by reducing the size of the raw materials. The pretreated biomass can then be processed using one or more of radiation exposure (under controlled temperature conditions), sonication, oxidation, pyrolysis, and steam explosion. Different systems and methods for pretreatment can use combinations of two, three or even four of these technologies.

Alternatively or additionally, the present invention is based, at least in part, on the observation that the methods described herein can be used to convert biomass into non-energy materials and compositions. Such materials and compositions include, but are not limited to, foodstuffs (eg, suitable for human and/or animal consumption), pharmaceuticals, nutraceuticals, pharmaceutical delivery vehicles and dosage forms, pharmaceutical excipients, pharmaceutical conjugates, cross-linked matrices , such as hydrogels, absorbent materials, fertilizers and lignin products. (516)

TYPES OF BIOMASS

Generally, any biomass material that is or includes carbohydrates that consist entirely of one or more saccharide elements or includes one or more saccharide elements can be processed by any of the methods described herein. As used herein, biomass includes cellulosic, hemicellulosic, starchy, and lignin-containing materials. For example, the biomass material may be cellulosic or lignocellulosic materials or starchy materials such as corn kernels, rice kernels or other feedstuffs, or materials that are or include one or more low molecular weight sugars such as sucrose or cellobiose.

For example, such materials may include paper, paper products, wood, wood-based materials, pressed wood, grasses, rice husks, cake, cotton, jute, hemp, flax, bamboo, sisal, abaca, straw, corncob cores, coconut fibers, algae, sea grass (eg, giant seaweed), water hyacinth, cassava, coffee beans, ground coffee beans (regular ground coffee beans), synthetic cellulose, or mixtures of any of these.

Fiber sources include cellulosic fiber sources, including paper and paper products (eg, laminated paper and kraft paper), and lignocellulosic fiber sources, including wood and wood-related materials, such as pressed wood. Other suitable fiber sources include natural fiber sources such as grasses, rice husks, rice husks, jute, hemp, flax, bamboo, sisal, abaca, straw, corncob cores, coconut fibers; sources of fibers with a high content of sa-cellulose, for example, cotton; and synthetic fiber sources such as extruded yarn (oriented yarn or non-oriented yarn). Natural or synthetic sources of fibers can be obtained from scraps of primary textile materials, such as scraps, or they can be used waste, such as rags. When paper products are used as fiber sources, they may be natural materials, such as scraps of virgin materials, or they may be used waste. In addition to primary raw materials, waste from used products, industrial (for example, offal) waste and processing waste (for example, disposable

From water from paper processing). Also, the source of the fibers can be obtained or formed from human waste (eg sewage), animals or plants. Additional fiber sources are described in the art, e.g., see US MoMo patents 6448307, 6258876, 6207729, 5973035 and 5952105.

Microbial sources include, but are not limited to, any naturally occurring or genetically modified microorganism and/or organism that contains or is capable of providing a source of carbohydrates (e.g., cellulose), such as unicellular organisms (e.g., animals (e.g., protozoa , such as flagellates, amoebae, ciliates and sporozoites) and plants (e.g. algae such as alveolobionts, chlorarachniophytes, cryptomonads, euglenids, glaucophytes, haplophytes, red algae, straminopyles and green algae)), sea grass, plankton (e.g. macroplankton , mesoplankton, microplankton, nanoplankton, picoplankton and femtoplankton), phytoplankton, bacteria (for example, gram-positive bacteria, gram-negative bacteria and extremophiles), yeast and/or their mixtures. In some cases, the microbial biomass can be obtained from natural sources, such as the ocean, lakes, bodies of water, such as salt water or fresh water, or from sources on land. Alternatively or additionally, microbial biomass can be obtained from culture systems, such as large-scale dry and wet culture systems.

Examples of biomass include renewable organic material such as plant biomass, microbial biomass, animal biomass (eg, any livestock by-product, livestock waste, etc.) and municipal waste biomass, including any and all combinations of these biomass materials. .

Plant biomass and lignocellulosic biomass include organic material derived from plants (woody or non-woody), especially material that is available on a permanent basis.

Examples include agricultural or food crop biomass (e.g., sugar cane, sugar beet, or corn kernels) or their extract (e.g., sugar from sugar cane and corn starch from corn), agricultural wastes and residues, such as corn straw, wheat straw, rice straw, sugarcane bagasse, cotton, etc. In addition, plant biomass includes, but is not limited to, trees, woody energy crops, wood waste and residues such as softwood chips, bark waste, wood sawdust, waste streams paper and pulp industry, wood fiber, etc., because as another source of plant biomass, forage grasses such as millet, etc. can potentially be grown on a large scale. For urban areas, the best potential plant biomass materials include horticultural waste (e.g. grass clippings, leaves, tree trimmings and windbreakers) and vegetable processing waste.

In some embodiments, the biomass includes lignocellulosic feedstock, which may be plant biomass, such as, but not limited to, non-woody plant biomass, crops such as, but not limited to, grasses, such as, but not limited to, C4- grasses such as millet, spartina, ryegrass, miscanthus, sedge or combinations thereof, or residues from sugar processing such as cake or beet pulp, agricultural residues such as soybean straw, corn straw, rice straw, rice husk, barley straw, pith corn cob, wheat straw, canola straw, oat straw, oat hulls, corn fiber, recycled wood pulp fiber, wood sawdust, hardwood such as aspen wood and sawdust, softwood, or combinations thereof. In addition, the lignocellulosic feedstock may include waste cellulosic materials such as, but not limited to, newsprint, cardboard, sawdust, etc. The lignocellulosic feedstock may include a single type of feedstock or, alternatively, the lignocellulosic feedstock may include a mixture of fibers that can come from different lignocellulosic raw materials. Moreover, the lignocellulosic feedstock may include fresh lignocellulosic feedstock, partially dried lignocellulosic feedstock, fully dried lignocellulosic feedstock, or a combination thereof.

Microbial biomass includes biomass that is derived from unicellular organisms and/or multicellular organisms that are naturally occurring or genetically modified, such as organisms from the ocean, lakes, bodies of water, such as salt water or fresh water, or terrestrial organisms, and contains a carbon source ( for example, cellulose). Microbial biomass may include, but is not limited to, e.g., unicellular organisms (e.g., animals (e.g., protozoa such as flagellates, amoebae, ciliates, and sporozoites)) and plants (e.g., algae such as alveolobionts, chlorarachniophytes, cryptomonads, euglenids, glaucophytes, haplophytes, red algae, straminopile and green algae)), sea grass, plankton (e.g. macroplankton, mesoplankton, microplankton, nanoplankton, picoplankton and femptoplankton), phytoplankton, bacteria (e.g. gram-positive bacteria,

Gram-negative bacteria and extremophiles), yeast and/or their mixtures. In some cases, the microbial biomass can be obtained from natural sources, such as the ocean, lakes, bodies of water, such as salt water or fresh water, or from sources on land. Alternatively or additionally, microbial biomass can be obtained from culture systems, such as large-scale dry and wet culture systems.

Animal biomass includes any organic waste material, such as animal-derived waste material or excrement, or human waste material or excrement (eg, compost and sewage).

In some embodiments, the carbohydrate is or includes a material that has one or more D-1,4 linkages and has a number average molecular weight of about 3,000 to 50,000. Such a carbohydrate is or includes cellulose (I) that is formed from D-glucose 1 by condensation of DV-(1--4)-glycosidic bonds. This bond is the opposite of a-(1--4)-glycosidic bonds present in starch and other carbohydrates. but po no he 1 he! ish (F) (Fin y o (F) y no 7 on on

AND

Starch materials include starch itself, such as corn starch, wheat starch, potato starch or rice starch, a starch derivative, or a material that includes starch, such as a food or crop. For example, the starch material may be arachis, buckwheat, banana, barley, cassava, kudzu, sorrel, sago, sorghum, common house potato, sweet potato, taro, yam, or one or more legumes such as horse beans, lentils, or peas. Also starchy materials are mixtures of these and/or other starchy materials. In specific embodiments, the starch material is derived from corn. Various corn starches and their derivatives are described in "Sogp Zagsp", Sogp Keipeg5 Azzosiayop (11th edition, 2006).

Biomass materials that include low molecular weight sugars may include, for example, at least about 0.5 wt.95 low molecular weight sugar, for example at least about 2, 3, 4, 5.6, 7, 8, 9, 10, 12.5, 25 , 35, 50, 60, 70, 80, 90 wt. 95 or even at least approximately 95 wt.9o low molecular weight sugar. In some cases, the biomass essentially consists of low molecular sugar, for example, more than 95 wt. 95, for example 96, 97, 98, 99 wt. 95 or essentially 100 wt. 95 low molecular sugar.

Biomass materials that include low molecular sugars can be agricultural or food products, such as sugar cane or sugar beet, or their extract, such as sugar cane or sugar beet juice. Biomass materials that include low molecular weight sugars can be essentially pure extracts, such as unrefined or crystallized table sugar (sucrose). Low molecular weight sugars include sugar derivatives. For example, low molecular weight sugars can be oligomeric (eg, equal to or greater than 4-mer, 5-mer, b6-mer, 7-mer, 8-mer, 9-mer, or 10-mer), trimeric, dimeric, or monomeric. When carbohydrates are formed by more than one repeating element, all the repeating elements can be the same or different.

Specific examples of low molecular sugars include cellobiose, lactose, sucrose, glucose and xylose, as well as their derivatives. In some cases, sugar derivatives dissolve more quickly in the solution or are utilized by microbes to produce useful material.

A few of these sugars and sugar derivatives are presented below.

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To produce any of the products described herein, it is possible to use combinations (eg, alone or in combination with any biomass material, component, product and/or co-product obtained using the methods described in this document) of any biomass materials described in this document. For example, mixtures of cellulosic materials and starch materials can be used to produce any product described herein.

BIOMASS PROCESSING SYSTEMS

In Fig. 1 presents a system for converting biomass 100, in particular biomass with significant amounts of cellulosic and lignocellulosic components and/or starch components, into useful products and by-products. System 100 includes a feedstock preparation subsystem 110, a pretreatment subsystem 114, a primary processing subsystem 118, and a further processing subsystem 122. Feedstock preparation subsystem 110 is fed raw biomass and physically prepared for use as raw materials for further processes (for example, size reduction and homogenization of biomass) and stored both in raw form and in the form of raw materials.

Biomass feedstocks with significant amounts of cellulosic and/or lignocellulosic components or starch components may have a high average molecular weight and crystallinity, which may complicate the processing of the feedstock into useful products (for example, fermentation of the feedstock to produce ethanol). Thus, it is useful to process biomass raw materials, for example, using the processing methods described in this document. As described herein, in some embodiments, biomass processing does not use acids, bases, and/or enzymes for biomass processing, or uses such processing only in small or catalytic amounts.

Biomass raw materials from the raw material preparation subsystem 110 are fed into the processing subsystem 114 and the raw materials are prepared in it for use in the main production processes,

for example, by reducing the average molecular weight and crystallinity of raw materials. From the processing subsystem 114, processed raw materials are fed to the primary processing subsystem 118 and useful products are produced in it (for example, ethanol, other alcohols, pharmaceuticals and/or food). In some cases, the product of the primary processing subsystem 118 is directly usable, but in other cases it requires additional processing performed by the downstream processing subsystem 122. The downstream processing subsystem 122 provides the downstream processing of the product stream from the primary processing system 118 that it requires (for example, distillation and denaturation of ethanol), as well as treatment of waste streams from other subsystems. In some cases, the co-products of the subsystems 114, 118, 122 may also be directly or indirectly suitable as secondary products iMabo to increase the overall efficiency of the system 100. For example, the subsystem for further processing 122 may produce treated water for recirculation as technical water in other subsystems, and/ or it can produce combustible waste that can be used as fuel for boilers that generate steam and/or electricity.

The optimal size of a biomass conversion plant is influenced by factors including economies of scale and the type and availability of biomass used as feedstock. Increasing plant size tends to increase the economies of scale associated with the processes in the plant. However, increasing plant size also tends to increase costs (such as transportation costs) per unit of raw material.

Studies analyzing these factors indicate that acceptable plant sizes for biomass conversion can range from 100 to 1,000 or more, such as 10,000 tons of dry feed per day, depending, at least in part, on the type of feedstock used. The type of biomass feedstock can also affect on-site storage requirements, where plants designed primarily to process feedstocks whose availability varies seasonally (e.g., corn straw) require more on-site than off-site storage of the feedstock. in comparison with installations designed for the processing of raw materials, the availability of which is relatively constant (for example, waste paper).

PRE-TREATMENT OF BIOMASS

In some cases, pretreatment methods begin with the physical preparation of biomass,

For example, reducing the size of biomass raw materials, for example, by slicing, grinding, grinding, crushing, crushing or chopping. In some embodiments, methods (for example, mechanical methods) are used to reduce the size or sizes of individual fragments of biomass. In some cases, loose raw materials (for example, recycled paper or millet) are pre-treated by crushing or cutting. Sieves and/or magnets can be used to remove very large objects or unwanted objects such as stones or nails from the feed stream.

Feedstock pretreatment systems can be tailored to produce raw material streams with specific characteristics, such as specific maximum dimensions, specific length-to-width ratios, or specific surface area ratios. As part of the preliminary processing of the raw material, the volume density of the raw material can be adjusted (for example, increased).

Size reduction

In some embodiments, the biomass is in the form of a fibrous material that includes fibers obtained by crushing the biomass. For example, crushing can be done using a cutting device with a rotating knife.

For example, and referring to Fig. 2, the source of biomass fiber 210 is crushed, for example in a cutting device with a rotating knife, to obtain a first fibrous material 212.

The first fibrous material 212 is passed through a first screen 214 having an average pore size of 1.59 mm or less (1/16 inch, 0.0625 inch) to produce a second fibrous material 216. If desired, the fiber source may be ground prior to crushing, for example using a shredder. For example, when paper is used as the fiber source, the paper may first be shredded into strips with a width of, for example, 1/4 to 1/2 inch (0.64 to 1.28 cm) using a shredding device, such as a shredder with counter-rotating screws, such as the device made by Mipsop ((isa, MM). As an alternative to shredding, the size of the paper can be reduced by cutting it to the desired size using a guillotine cutter. For example, a guillotine cutter can be used to cut paper into sheets , for example, 10 inches (25.4 cm) wide and 12 inches (30.5 cm) long.

In some embodiments, crushing the fiber source and passing the resulting first fibrous material through the first sieve are performed simultaneously. Crushing and passing can also be carried out in a sequential process.

For example, a cutting device with a rotating knife can be used to simultaneously crush the fiber source and screen the first fibrous material. Referring to

Fig. 3, the rotary knife cutting device 220 includes a hopper 222 into which the shredded fiber source 224 obtained by standard methods can be placed. The chopped fiber source is crushed between stationary blades 230 and rotating blades 232 to produce a first fibrous material 240. The first fibrous material 240 is passed through a sieve 242 and the resulting second fibrous material 244 is collected in a basket 250. To facilitate the collection of the second fibrous material, the basket may be pressurized below nominal atmospheric pressure, such as at least 1095 below nominal atmospheric pressure, such as at least 25 95 below nominal atmospheric pressure, at least 50 95 below nominal atmospheric pressure, or at least 75 95 below nominal atmospheric pressure. In some embodiments, a vacuum source 252 is used to maintain basket pressure below nominal atmospheric pressure.

Crushing can be beneficial for "opening up" and "stressing" fibrous materials, making the cellulose of the materials more susceptible to chain separation and/or reduced crystallinity. Exposed materials may also be more sensitive to oxidation when exposed to radiation.

In some embodiments, crushing may be preferred for "opening up" and "stressing" the fibrous materials, making the cellulose of the materials more susceptible to breakdown and absorption in ruminants.

The fiber source can be crushed in a dry state, in a hydrated state (for example, having up to ten percent by weight of absorbed water) or in a wet state, for example, having from about 10 wt. 95 to about 75 wt. 95 water. The fiber source can be crushed even when partially or fully immersed in a liquid such as water, ethanol, or isopropanol.

The fiber source can also be crushed in a gas atmosphere (such as a stream or atmosphere of a gas other than air), such as oxygen or nitrogen or steam.

Other methods of obtaining fibrous materials include, for example, mill grinding, mechanical fluffing or tearing, rod mill grinding, or air grinding.

If desired, the fibrous materials can be separated, for example continuously or in batches, into fractions according to their length, width, density, type of material, or some combination of these characteristics.

For example, ferrous metals can be separated from any fibrous material by passing the fibrous material, which includes the ferrous metal, past a magnet, such as an electromagnet, and then passing the resulting fibrous material through a series of sieves, each sieve having holes of a distinct size.

Fibrous materials can also be separated, for example, using a high-velocity gas such as air. In this approach, fibrous materials are separated by removing different fractions, which, if desired, can be subjected to photon characterization. Such a device for separation is considered in the first page of the article, US patent No. 6883667.

The fibrous materials can be pretreated immediately after preparation, or they can be dried, for example, at about 105 °C for 4-18 hours, so that the moisture content before use is, for example, less than about 0.5 95.

If desired, the lignin can be removed from fibrous materials that include lignin. Also, to facilitate degradation of materials that include cellulose, the material may be treated prior to irradiation with heat, a chemical reagent (eg, a mineral acid, a base, or a strong oxidizing agent such as sodium hypochlorite), and/or an enzyme.

In some embodiments, the average opening size of the first screen is less than 0.79 mm (1/32 inch, 0.03125 inch), such as less than 0.51 mm (1/50 inch, 0.02000 inch), less than 0.40 mm (1/64 in, 0.015625 in), less than 0.23 mm (0.009 in), less than 0.20 mm (1128 in, 0.0078125 in), less than 0.18 mm (0.007 in), less than 0, 13 mm (0.005 in) or even less than 0.10 mm (1/256 in, 0.00390625 in). The sieve is made by interweaving monofilament, which has the appropriate diameter to obtain the desired hole size. For example, monofilaments can be made of metal such as stainless steel. As the size of the holes decreases, the structural requirements for the monofilament increase. For example, for hole sizes less than 0.40 mm, it may be preferable to fabricate monofilament screens made from a material other than stainless steel, such as titanium, titanium alloys, amorphous metals, nickel, tungsten, rhodium, rhenium, ceramics, or glass. In some embodiments, the sieve is made from a plate, such as a metal plate, having holes, for example, cut into the plate using a laser. In some embodiments, the area of the holes in the sieve is less than 52 95, for example less than 41 95, less than 36 95, less than 31 95, less than 30 95.

In some embodiments, the second fibrous material is crushed and passed through a first sieve or sieve with a different size. In some embodiments, the second fibrous material is passed through a second sieve having an average pore size equal to or less than the pore size of the first sieve.

Referring to FIG. 4, the third fibrous material 220 can be obtained from the second fibrous material 216 by crushing the second fibrous material 216 and passing the resulting material through a second sieve 222 having an average pore size smaller than that of the first sieve 214.

Typically, the fibers of fibrous materials can have a relatively high average length to diameter ratio (eg, greater than 20 to 1), even if they have been subjected to crushing more than once. In addition, the fibers of the fibrous materials described herein may have a relatively narrow length distribution and/or length-to-diameter ratio.

As used herein, the average fiber width (eg, diameter) is determined optically by randomly selecting approximately 5,000 fibers. Average fiber length is length-weighted adjusted lengths. Surface area of VET (Vhypaneg,

Ettei and TeIPeg) is a multipoint surface area, and porosity is a quantity determined by mercury porometry.

The average ratio of the length to the diameter of the second fibrous material 14 can be, for example, more than 5/1, more than 8/1, for example, more than 10/1, more than 15/11, more than 20/1, more than 25/1 or even more than 50/1. The average length of the second fibrous material 14 may be, for example, about 0.5 to 2.5 mm, such as about 0.75 to 1.0 mm, and the average width (ie, diameter) of the second fibrous material 14 may be, for example, from about 5 to 50 µm, for example from about 10 to 30

MKM.

In some embodiments, the standard deviation of the length of the second fibrous material 14 is less than 60 95 from the average length of the second fibrous material 14, such as less than 50 95 from the average length, less than 40 95 from the average length, less than 25 95 from the average length, less than 10 95 from the average length, less than 5 95 of the average length or even less than 1 95 of the average length.

In some embodiments, the VET surface area of the second fibrous material is greater than 0.1 mg/g, such as greater than 0.25 m-/g, greater than 0.5 mg/g, greater than 1.0 mg/g, greater than 1.5 m- /g, exceeds 1.75 m-/g, exceeds 5.0 m-/g, exceeds 10 m-/g, exceeds 25 me/g, exceeds 35 m"/g, exceeds 50 m"/g, exceeds 60 m"/g, exceeds 75 m"/g, exceeds 100 me/g, exceeds 150 mg/g, exceeds 200 m"/g or even exceeds 250 m"/g. The porosity of the second fibrous material 14 may, for example, exceed 20 95, Exceed 25 95, Exceed 35 95, Exceed 50 95, Exceed 60 95, Exceed 70 95, Exceed 80956, Exceed 85905, Exceed 9095, Exceed 92 95, Exceed 94 95, Exceed 95 95, Exceed 97.5 95, exceed 99 95 or even exceed 99.5 95.

In some embodiments, the ratio of the average length-to-diameter ratio of the first fibrous material to the average length-to-diameter ratio of the second fibrous material is, for example, less than 1.5, for example, less than 1.4, less than 1.25, less than 1.1, less than 1.075, less than 1.05, less than 1.025, or even essentially equal to 1.

In specific embodiments, the second fibrous material is crushed again and the resulting fibrous material is passed through a second sieve having an average pore size smaller than that of the first sieve to produce a third fibrous material. In such cases, the ratio of the average length-to-diameter ratio of the second fibrous material to the average length-to-diameter ratio of the third fibrous material may be, for example, less than 1.5, such as less than 1.4, less than 1.25, or even less than 1.1.

In some embodiments, the third fibrous material is passed through a third sieve to produce a fourth fibrous material. The fourth fibrous material can, for example, be passed through the fourth sieve to obtain the fifth material. Similar sieving processes can be repeated as many times as desired to obtain the desired fibrous material having the desired properties. because Sealing

As used in this description, densification refers to an increase in the bulk density of a material. Compacted materials may be recycled, or any recycled materials may be compacted in any of the ways described herein.

A material such as a fibrous material having a low bulk density can be compacted into a product having a higher bulk density. For example, a composition of material having a bulk density of 0.05 g/cm can be compacted by isolating the fibrous material in a relatively gas-tight structure, such as a bag made of polyethylene or a bag made of alternating layers of polyethylene and nylon, with further removal from the structure of the gas contained in it, such as air. After the air is removed from the structure, the fibrous material may have, for example, a bulk density greater than 0.3 g/cmU, such as 0.5, 0.6, 0.7 g/cm3 or greater, such as 0.85 g/cm3. After compaction, the product can be pretreated by any of the methods described in this document, such as irradiation, such as gamma radiation. This can be advantageous when it is desired to transport the material to another location, such as a remote manufacturing facility, where the fibrous material composition can be added to a solution, for example, to produce ethanol. After piercing the substantially gas-impermeable structure, the densified fibrous material may return to substantially its original bulk density, such as at least 60 95 of its original bulk density, such as 70, 80, 85 95 or more, such as 95 95 of its original volume of capacitive density. To reduce static electricity in the fibrous material, a means that removes static charges can be added to the material.

In some embodiments, the structure, such as a carrier such as a bag, is made of a material that dissolves in a liquid such as water. For example, the structure can be made of polyvinyl alcohol so that it dissolves when in contact with an aqueous solution. Such variants of implementation allow adding compacted structures directly into solutions that include a microorganism, without primary release of the contents of the structure, for example, by cutting.

Referring to FIG. 5, the biomass material can be combined with any desired additives and binder and then compacted by applying pressure, for example by passing the material through a certain gap between counter-rotating pressure rollers

By or by passing the material through a granulation press. During the application of pressure, heating may optionally be applied to facilitate compaction of the fibrous material. The compacted material can then be irradiated.

In some embodiments, the material before compaction has a bulk density of less than 0.25 g/cm3, such as less than 0.20, 0.15, 0.10, 0.05 g/cm" or less, such as 0.025 g/cm3. Bulk density is determined using ATM 018958. Briefly, the method involves filling a measuring cylinder with a known volume of sample and determining the mass of the sample.The bulk density is calculated by dividing the mass of the sample in grams by the known volume of the cylinder in cubic centimeters.

Preferred binders include binders that are water-soluble, water-swellable, or that have a glass transition temperature of less than 25°C, as determined by differential scanning calorimetry. Water-soluble binders have solubility in to water at least about 0.05 wt. 95. Water-swellable binders are binders whose volume increases by more than 0.5 wt. 95 under the influence of water.

In some embodiments, binders that are water-soluble or water-swellable include functional groups that are capable of forming a bond, such as a hydrogen bond, with the fibers of a fibrous material, such as a cellulose fibrous material. For example, a functional group can be a carboxylic acid group, a carboxylate group, a carbonyl group, such as an aldehyde or a ketone, a sulfonic acid group, a sulfonate group, a phosphoric acid group, a phosphate group, an amide group, an amino group, a hydroxyl group, such as an alcohol, and combinations thereof. groups, for example, a carboxylic acid group and a hydroxyl group. Specific examples of monomers include glycerol, glyoxal, ascorbic acid, urea, glycine, pentaerythritol, monosaccharide or disaccharide, citric acid and tartaric acid. Suitable saccharides include glucose, sucrose, lactose, ribose, fructose, mannose, arabinose and erythrose. Examples of polymers include polyglycols, polyoxyethylene, polycarboxylic acids, polyamides, polyamines and polysulfonic acids, polysulfonates. Specific examples of polymers include polypropylene glycol (PPO), polyethylene glycol (PES), polyoxyethylene, e.g., ethylene oxide and propylene oxide copolymers, polyacrylic acid (PAA), polyacrylamide, polypeptides, polyethyleneimine,

polyvinylpyridine, poly(sodium-4-styrenesulfonate) and poly(2-acrylamidomethyl-1-propanesulfonic acid).

In some embodiments, the binder includes a polymer that has a glass transition temperature of less than 25°C. Examples of such polymers include thermoplastic elastomers (TPEs). Examples of TPEs include polyester block amides such as polyester block amides, available under the trade name REVAHF), polyester elastomers such as polyester elastomers available under the trade name NUTREIG FU, and styrenic block copolymers such as block copolymers available under the trade name KKATOMF).Other suitable polymers having a transition temperature of glassy state less than 25 "C, include copolymer of ethylene and vinyl acetate (EMA), polyolefins, such as polyethylene, polypropylene, copolymers of ethylene-propylene, and copolymers of ethylene and alpha-olefins, such as 1-octene, such as copolymers available under the trade name EMOSACEF. In some embodiments, for example, when the material is pulped paper with a multilayer coating, the material is compacted without adding a special polymer with a low glass transition temperature.

In a specific embodiment, the binder is lignin, for example natural or synthetically modified lignin.

A suitable amount of binder added to the material, calculated on a dry weight basis, is, for example, from about 0.01 95 to about 50 95, for example 0.03, 0.05, 0.1, 0, 25, 0.5, 1.0, 5, 10 95 or more, for example 25 95, based on the total mass of compacted material. The binder can be added to the material as an undiluted pure liquid, as a liquid in which the binder is dissolved, as a dry binder powder, or as granules of the binder.

Compacted fibrous material can be made in a pelletizing press.

Referring to FIG. 6, the pelletizing press 300 has a hopper 301 for holding uncompacted material 310, which includes carbohydrate-containing materials such as cellulose. The container is connected to an auger 312 which is driven by a variable speed motor 314 so that the uncompacted material can be transported to a mixing device 320 in which the uncompacted material is mixed by blades 322 which are rotated by the motor.

Out of 330 mixing devices. Other ingredients, such as any additives and/or fillers described herein, may be added through inlet 332. If desired, heat may be added while the fibrous material is in the mixing device. After mixing, the material is removed from the mixing device through a sleeve 340 to another auger 342. The sleeve, controlled by the drive mechanism 344,

C5 allows the unhindered passage of material from the mixing device to the auger.

The screw rotates under the action of motor 346 and controls the supply of fibrous material to the die assembly with rollers 350. Specifically, the material is fed into a hollow cylindrical die 352 which rotates about a horizontal axis and which has radial holes 250. The die 352 is rotated about an axis under the action of motor 360, which includes a power meter that indicates the total energy consumed by the engine. Compacted material 370, for example, in the form of pellets, falls from the chute 372 and is collected and processed, for example, by irradiation.

It is convenient that after compaction the material has the form of granules or chips that take different shapes. Then the pellets can be irradiated. In some embodiments, the granules or chips are cylindrical in shape, for example having a maximum transverse dimension of, for example, 1 mm or more, for example 2, 3, 5, 8, 10, 15 mm or more, for example 25 mm. Other convenient forms include pellets or chips that are plate-shaped, for example having a thickness of 1 mm or more, for example 2, 3, 5, 8, 10 mm or more, for example 25 mm; width, for example, 5 mm or more, for example 10, 15, 25, 30 mm or more, for example 50 mm; and

BO length of 5 mm or more, for example 10, 15, 25, 30 mm or more, for example 50 mm.

Next, referring to FIG. 7A-70, the pellets can be made to have a cavity inside. As shown, the cavity can be located mainly in line with the center of the pellet (Fig. 7B) or it can be offset from the center of the pellet (Fig. 7C).

Making the pellet hollow inside can increase the dissolution rate in the liquid after irradiation.

Next, referring to FIG. 70, the pellet may have, for example, a cross-sectional shape that is multilobed, such as three-lobed as shown, or four-lobed, five-lobed, six-lobed or ten-lobed. Making granules with this transverse shape can also increase the dissolution rate in the solution after irradiation.

Alternatively, the compacted material may have any other desired shape, for example the compacted material may be in the form of a plate, cylinder or briquette.

Examples of compaction

In one example, half-gallon (1.9 L) juice cartons made from white kraft paperboard with a bulk density of 20 lb/ft can be used as raw materials? (0.32 g/cmU). The paperboard can be folded flat and then fed into a shredder to produce a confetti-like material that is 0.1 inch (0.25) to 0.5 inch (1.27 cm) wide, 0. 25 inches (0.63 cm) to 1 inch (2.54 cm) and a thickness equivalent to the thickness of the parent material (approximately 0.075 inches (0.19 cm)). The confetti-like material can be fed into a cutting device with a rotating knife that crushes the confetti-like fragments, breaking the fragments and releasing the fibrous material.

In some cases, several shredder-shredder systems can be arranged with sequential production. In one embodiment, two shredder-shredder systems may be arranged in series, where the product of the first shredder is fed as input to the second shredder. In another embodiment, the three crushing device-crushing device systems can be arranged in series, where the product of the first crushing device is fed as input to the second crushing device and the product of the second crushing device is fed as input to the third crushing device. It is expected that multiple passes through crusher-crusher systems can reduce particle size and increase the total surface area in the feed stream.

In another example, fibrous material obtained by crushing and crushing juice cartons can be processed to increase its bulk density. In some cases, the fibrous material can be sprayed with water or diluted mother liquor

ROIGMOH'"M UuUBE M10 (polyoxyethylene) prepared in water. The soaked fibrous material can then be processed in a granulation press operating at room temperature.

A granulation press can increase the volume density of the source material flow by more than one order of magnitude.

Co.) PROCESSING

Pretreated biomass can be processed for use in the main production processes, for example, by reducing the average molecular weight, crystallinity and/or increasing the surface area and/or porosity of the biomass. In some embodiments, the biomass can be treated to reduce the recalcitrance of the biomass. Processing processes may include at least one (eg, one, two, three, four or five) of irradiation, sonication, oxidation, pyrolysis and steam explosion.

Resilience is a term used in the art, as used herein, that broadly refers to the resistance of a biomass material to polysaccharide-degrading agents (e.g., microorganisms and/or enzymes (e.g., microbial enzymes)) from accessing the polysaccharides contained in the biomass (see, for example, Nittei ei ai.,

Mayiope! Nepezhabiye Epegda I arogayuga (MAEG) TesNpis! Neroi MAES/TR-510-37902, A!ydivi, 2005 and Mayopa! Kepejarie Epegdu Garogaigu (MKE!) Tesppis! Kerogi MKEI/VK-510-40742, Magsy, 2007). For example, the availability of polysaccharides (eg, cellulose and hemicellulose) in the first biomass material with the first level of recalcitrance is lower than the availability of polysaccharides (eg, cellulose and hemicellulose) in the same lignocellulosic material after treatment to reduce the level of recalcitrance of the material. In other words, the level of polysaccharides available to polysaccharide-degrading agents is higher after treatment to reduce recalcitrance.

Assessment of levels of recalcitrance of lignocellulosic biomass

The level of recalcitrance of lignocellulosic material can be assessed using a number of methods known in the field. Examples of such methods include, but are not limited to, surface characterization methods, enzymatic methods, and functional methods.

Illustrative methods of surface characterization that can be used to assess the level of recalcitrance of lignocellulosic materials are known in the art (for a review, see Nittei et al.,

Mayiope! Nepezhabie Epegdu And Aroghaig (MAEIG) Tesnpis! Heroi MAEK/TR-510-37902, A!ydivi, 2005 and Oipd ei ai, Misgozsoru apa Misgoapaiuzi5, 14:1494-1495, 2004). For example, the level of refractoriness of lignocellulosic materials can be assessed using microscopic or spectroscopic methods of surface analysis (e.g., using one or more of the surface analysis methods described below) to identify, evaluate, and/or quantify changes (e.g., structural changes) in lignocellulosic materials. materials, which indicate a decrease in the inflexibility of the material. Illustrative changes that can be used as signs of reduced refractoriness in lignocellulosic materials include the appearance of pits or pores, and/or the surface of unfolded microfibrils. See, for example, Nitythei ei ai!., Mayopa! Hats off

Epegau Iarogaitu (MAE!) Tesppisai Veroiy MAE /TR-510-37902, A!ydivi, 2005 and Oipd ei ai.,

Mistozsoru apa Mistoapa|tsuviv, 14:1494-1495, 2004, where the following methods are described: (1) Scanning electron microscopy (SEM) can be used to visualize the surface morphology of biological and non-biological materials in a wide range of magnifications (magnification up to 20000X) and with high depth of field (see, for example, sote? ei a!., VibtesppoYodu

Tog Vioishei!5, 1, Osioreg 23, 2008; 5imap ei a!., Arri. Misgobrio!. Viotesppo!., 72:346-352, 2006). Typically, biological samples, such as lignocellulosic biomass samples, are coated with a thin layer of an electron-dense material such as carbon or atomized gold prior to analysis.

For example, samples can be placed in 5EM couplings and coated with gold/palladium. These placed samples can then be observed using known methods and devices, e.g

UEOG OM 69401 M BEM (Cheoi Pa., ToKuo, Shdarap), with an accelerating voltage of 5 kV. (2) Later, methods were developed to analyze samples containing natural moisture, a method called environmental mode EEM (EMEM), for example using

Otsapia HPP 400 EBEM (RE! Sotrapu). The use of E5EM in the analysis of yeast cells is described

Vep ei ai., Ipmeviidayop oi She togrpoiodu, miariyu apa tespapisa! rgorepie5z oi uyeavi seiYiv ip epmigoptepiai! 5EM, 5sappipo, ribiiipei opiipe A!ydivi 5, 2008). Such methods in the mode of the natural environment can be used for the analysis of lignocellulosic biomass containing moisture, without the use of electron-dense coatings. (3) Atomic force microscopy (AEM) can also be used, for example using OI-Meeso MiyiMode RisoBogse 5uziet (see, for example, Zytsed ei ai., Chem. 5061.

Ipzigit., 79:103701, 2008). AEM effectively allows the analysis of surface topography at very high magnification, while also allowing the analysis of attractive and repulsive forces between the tip of the scanning probe and the sample surface, thus providing a height image and a phase image. AEM is increasingly used for the analysis of biological samples due to its high atomic level of resolution and its ease of use (samples do not require lengthy preparation). In addition, AEM can be used to monitor

With dry and hydrated surfaces directly using a tapping probe. (4) Transmission Electron Microscopy (TEM), for example, using the EEE Tespai E20, allows determination of the internal structures of biological and non-biological materials up to a magnification of at least 350,000X. As a rule, the definition of internal structures can be facilitated by using shadow methods of imaging or staining with high-contrast compounds. It is also possible to carry out compositional analysis of materials by monitoring secondary X-rays produced during electron-sample interaction using energy-dispersive X-ray microanalysis. TEM-based methods for analyzing the level of recalcitrance of lignocellulosic material are described in the art (see, for example, Kpoadvs ei a!., Sap. 9. Misgobioi, 41:592-600, 1995). (5) Near-field optical microscopy (NEMOM) using, for example, OI-Aigoga-3

The MOM (Mikop) enables the observation of surfaces using a light microscope with a long depth of field, which is adapted to perform secondary spectrophotometric analysis, such as CML/LZ, fluorescence analysis and laser Raman analysis. In some variants, MEBOM can be carried out using an OIutry5 1 x 71 inverted microscope equipped with a CSO camera with a high resolution ОР7О, for microscopy of individual molecules. (6) Confocal microscopy (SEM) and confocal scanning laser microscopy (SIM) (see, for example, Mayopa! Kepemjarie Epegdu Garogaigu (MKEM) Tesppis! Keropgyi MKES/VK-510-40742, Magsp, 2007) can be used to obtain optical sections that can be used to construct a three-dimensional image of the surface and internal structures. As a rule, SEM and C5BIM are carried out in combination with methods of labeling, for example, with fluorescent dyes (see, for example, boie ei ai., Mishor. Esoi., Rybiyepei opiipe op

Mometreg 4, 2008).

In some embodiments, the level of recalcitrance of the lignocellulosic material can be assessed using one or more methods known in the art, such as methods described herein. The same sample or portion thereof can then be evaluated after processing to detect a change (eg, a structural change) in refractoriness. In some embodiments, the appearance or observation of dimples or pores, and/or the surface spread of microfibrils in or on the first lignocellulosic material with the first level of refractoriness is less than the appearance or observation of pits or pores, and/or the surface spread of microfibrils in the sample after treatment for reducing the level of inflexibility of the material.

Alternatively or additionally, a change (for example, a decrease) in the level of recalcitrance of the lignocellulosic material can be analyzed using enzymatic methods.

For example, lignocellulosic material can be incubated in the presence of one or more cellulases, for example, before or after treatment using methods described herein. In some embodiments, an increase in the destruction of cellulose by cellulase indicates a change in the level of refractoriness of the material, such as a decrease in the refractoriness of the material. In some embodiments, increasing the destruction of cellulose by cellulase leads to an increase in the amount of monosaccharides and/or disaccharides in the sample.

In some embodiments, the amount (e.g., concentration) of monosaccharides and/or disaccharides due to enzyme (e.g., cellulase) activity in the sample containing the first lignocellulosic material with the first level of recalcitrance is lower than the amount (e.g., concentration) of monosaccharides and/or disaccharides. or disaccharides due to the activity of an enzyme (eg, cellulase) in the same sample after treatment to reduce the level of recalcitrance of the material.

Alternatively or additionally, a change (eg, a decrease) in the level of recalcitrance of the lignocellulosic material can be analyzed using functional methods.

For example, lignocellulosic material can be cultivated in the presence of a fermenting sugar microorganism, for example, using the methods of cultivation described herein, before and after processing using the methods described herein. In some embodiments, an increase in the level of one or more products produced by the microorganism indicates a change in the level of refractoriness of the material, such as a decrease in the refractoriness of the material.

In some embodiments, the rate of growth of a microorganism and/or production of a product by a microorganism in a sample containing a first lignocellulosic material with a first level of refractoriness is lower than the rate of growth of a microorganism and/or production of a product by a microorganism in the same sample after a reduction in the level of refractoriness of the material.

In some embodiments, the change in material refractoriness level can be expressed as (1)

From the ratio (for example, the indicator of the level of material inflexibility before processing relative to the level of material inflexibility after processing); (2) a percentage change (for example, a decrease) in the material's recalcitrance level; (3) the percentage change (eg, decrease) in the level of polysaccharide available to the polysaccharide-degrading agent (eg, enzyme) after treatment, compared to the level before treatment, per unit mass of the biomass source material; or (4) a percentage change (eg, increase) in the solubility of the material in a particular solvent.

In some cases, the second material contains cellulose that has a crystallinity (Cg) that is lower than the crystallinity (Si) of the cellulose of the first material. For example, (Cg) may be lower than (Si) by more than about 10 95 , such as 15, 20, 25, 30, 35, 40 95 , or even more than about 50 95 .

In some embodiments, the initial crystallinity index (before irradiation) is from about 40 95 to about 87.5 95, such as from about 50 95 to about 75 95 or from about 60 95 to about 70 905, and the crystallinity index after irradiation is from about 10 95 to about 50 95, such as from about 15 95 to about 45 95 or from about 20 956 to about 40 95. However, in some embodiments, such as after extensive irradiation, the crystallinity index may be less than 5 95. In some embodiments implementation of the material after irradiation is essentially amorphous.

In some embodiments, the starting number average molecular weight (before irradiation) is from about 200,000 to about 3,200,000, such as from about 250,000 to about 1,000,000 or from about 250,000 to about 700,000, and the number average molecular weight after irradiation is from about 50,000 to about 200,000, e.g. from about 60,000 to about 150,000 or from about 70,000 to about 125,000. However, in some embodiments, such as after extensive irradiation, the number average molecular weight may be less than about 10,000 or even less than about 5,000.

In some embodiments, the second material may have an oxidation level (702) greater than the oxidation level (7101) of the first material. A higher level of oxidation of a material can contribute to its ability to disperse, swell and/or dissolve, further increasing the susceptibility of the material to chemical, enzymatic or biological attack. In some embodiments, to increase the oxidation level of the second material relative to the first material, the irradiation is carried out in an oxidizing atmosphere, such as air or oxygen, producing a second material that is more oxidized than the first material.

For example, the second material may have more hydroxyl groups, aldehyde groups, ketone groups, ester groups, or carboxylic acid groups that may increase its hydrophilicity.

Combined processing

In some embodiments, the biomass can be processed using at least one (e.g., two, three, four, or five) of the processing methods described herein, such as two or more of radiation exposure, sonication, oxidation, pyrolysis, and steam explosion, or with or without prior, intermediate or subsequent biomass preparation as described in this document. Treatment methods can be applied to biomass, such as cellulosic and/or lignocellulosic material, in any order, multiple times (eg, two or more applications of the treatment method), or simultaneously. In other embodiments, materials that include a carbohydrate are prepared using three, four, or more of any of the methods described herein (in any order or simultaneously). For example, a carbohydrate can be obtained by applying to cellulosic and/or lignocellulosic material radiation exposure, sonication, oxidation, pyrolysis, and, optionally, steam explosion (in any order or simultaneously). The resulting carbohydrate-containing material can then be converted by one or more microorganisms, such as bacteria (eg, gram-positive bacteria, gram-negative bacteria, and extremophiles), yeast, or a mixture of yeast and bacteria, into a number of desired products as described herein. Complex methods can provide materials that can be more easily utilized by various microorganisms due to their lower molecular weight, lower crystallinity and/or increased solubility. Complex methods can provide synergy and can reduce the total required energy costs, compared to any single method.

For example, in some embodiments, a biomass feedstock may be provided that includes a carbohydrate produced by a method that includes irradiating and sonicating (in any order or simultaneously) the biomass material by a method that includes irradiating and oxidizing (in any order or simultaneously) of biomass material, in a way that includes

From irradiation and pyrolysis (in any order or simultaneously) of biomass material, by a treatment method that includes irradiation and pyrolysis (in any order or simultaneously) of biomass material, or by a method that includes irradiation and steam explosion (in any order or simultaneously) of biomass material. Then the given raw biomass can be brought into contact with a microorganism capable of converting at least a portion, for example at least about 1 wt. 95, biomass into a product.

In some embodiments, the method does not include hydrolysis of the biomass, for example, with an acid, a base, and/or an enzyme, for example, a mineral acid such as hydrochloric or sulfuric acid.

If desired, a portion of the biomass may or may not include hydrolyzed material. For example, in some embodiments, at least about seventy percent by weight of the biomass is non-hydrolyzed material, such as at least 95 wt. 96 raw materials are non-hydrolyzed material. In some embodiments, substantially all of the biomass is unhydrolyzed material. In some embodiments, 100 95 of the biomass is non-hydrolyzed material.

Any raw material or any reactor or fermenter loaded with the raw material may include a buffer such as sodium bicarbonate, ammonium chloride, or Ti5; an electrolyte such as potassium chloride, sodium chloride or calcium chloride; a growth factor such as biotin and/or a base pair such as uracil or its equivalent; a surfactant such as Tueepo or polyethylene glycol; a mineral such as calcium, chromium, copper, iodine, iron, selenium or zinc; or a chelating agent such as ethylenediamine, ethylenediaminetetraacetic acid (EDTO) (or a salt form thereof, such as sodium or potassium EDTO) or dimercaprol.

When radiation exposure is used, it can be applied to any sample that is dry or wet, or even dispersed in a liquid such as water. For example, irradiation can be performed on biomass material having less than about 25 Mab. 95 biomass material has a surface wetted by a liquid such as water. In some embodiments, the irradiation is performed on a biomass material in which the biomass material is not substantially wetted by a liquid such as water.

In some embodiments, any processing described herein is performed on biomass material that remains dry as received or after drying the material, for example using heat and/or reduced pressure. For example, in some embodiments, the biomass material has less than about five percent by weight of retained water, measured at 25 "C and fifty percent relative humidity.

If desired, in any method described herein, a swelling agent as defined herein may be used. In some embodiments, when cellulosic and/or lignocellulosic material is processed using radiation exposure, less than about 25 wt. 96 of the biomass material is in a swollen state, where the swollen state is characterized as having a volume that is more than 2.5 95 greater than the volume in the non-swollen state, for example more than 5.0, 7.5, 10 or 15 95 exceeds the volume in the unswollen state In some embodiments, when radiation exposure is applied to the biomass material, the biomass material is essentially not in a swollen state.

In specific embodiments, when radiation exposure is used, the biomass material includes a swelling agent, and the swollen biomass material receives a dose of less than about 10 Mrad.

When radiation exposure is used in any method, it can be applied by simultaneously exposing the biomass to air, oxygen-enriched air or even oxygen, or in an inert gas atmosphere such as nitrogen, argon, or helium. When maximum oxidation is desired, an oxidizing medium such as air or oxygen is used.

When radiation exposure is used, it can be applied to biomass, such as cellulosic and/or lignocellulosic material, at pressures greater than about 2.5 atmospheres (253 kPa), such as greater than 5 (506 kPa), 10 (1012 kPa), 15 (1518 kPa), 20 (2036 kPa), or even more than about 50 atmospheres (5060 kPa). Irradiation can increase the solubility, swelling capacity, or dispersibility of the biomass in the solvent.

In specific embodiments, the method includes irradiation and sonication, and the irradiation precedes the sonication. In other specific embodiments, sonication precedes irradiation, or irradiation and sonication are performed essentially simultaneously.

In some embodiments, the method includes irradiation and ultrasound treatment (in any

In which order or at the same time) and additionally includes oxidation, pyrolysis or steam explosion.

When the method includes radiation exposure, the exposure may be performed using ionizing radiation such as gamma rays, an electron beam, or ultraviolet C radiation having a wavelength of from about 100 nm to about 280 nm, a particle beam, such as an electron beam, slow neutrons or alpha particles. In some embodiments, the implementation of irradiation includes two or more sources of radiation, such as gamma rays and an electron beam, which can be applied in any order or simultaneously.

In specific embodiments, sonication can be performed at a frequency of from about 15 kHz to about 25 kHz, such as from about 18 to 22 kHz, using a horn of 1 kW or more, such as a horn of 2, 3, 4, 5 kW, or even 10 kW

In some embodiments, the biomass has a first number average molecular weight, and the resulting carbohydrate includes a second cellulose having a second number average molecular weight lower than the first number average molecular weight. For example, the second number average molecular weight is lower than the first number average molecular weight by more than about twenty-five percent, for example the reduction is 2x, 3x,

Bh, 7x, 10x, 25x or even 100x.

In some embodiments, the first cellulose has a first crystallinity and the second cellulose has a second crystallinity lower than the first crystallinity, such as more than about two, three, five, ten, fifteen, or twenty-five percent lower crystallinity .

In some embodiments, the first cellulose has a first oxidation level and the second cellulose has a second oxidation level that is higher than the first oxidation level, such as two, three, four, five, ten, or even twenty-five percent higher.

In some embodiments, the first biomass has a first level of refractoriness and the final biomass has a second level of refractoriness that is lower than the first level.

Radiation treatment

For the processing of biomass from a wide variety of different sources, one or more processing sequences can be used for the purpose of extracting useful substances from the raw material and for the purpose of obtaining partially degraded organic material, which performs the function of an input stream in further processing stages and/or sequences. Irradiation can reduce the refractoriness, molecular weight and/or crystallinity of the raw material.

In some embodiments, materials are irradiated using energy stored in the material, which releases an electron from its atomic orbital. Radiation exposure can be provided by 1) heavy charged particles such as alpha particles or protons, 2) electrons produced, for example, in beta decay or in electron beam accelerators, or 3) electromagnetic radiation, such as gamma rays, X-rays or ultraviolet rays. In one approach, radiation generated by radioactive substances can be used to irradiate raw materials. In some embodiments, any combination of (1)-(3) can be used in any order or simultaneously. In another approach, electromagnetic radiation (for example, generated using electron beam sources) can be used to irradiate raw materials. The doses used depend on the desired effect and the specific raw material. For example, high doses of radiation exposure can destroy chemical bonds in the components of the raw material, and low doses of radiation exposure can increase the formation of chemical bonds (eg, cross-linking) in the components of the raw material. In some cases where chain separation is desired and/or functionalization of polymer chains is desired, particles heavier than electrons can be used, such as protons, a group of helium nuclei, argon ions, silicon ions, neon ions, carbon ions, phosphorus ions, oxygen ions, or nitrogen ions. When ring-opening chain separation is desired, positively charged particles can be used for enhanced ring-opening due to their Lewis acid properties.

Referring to FIG. 8, in one method, the first material 2, which is or includes cellulose having a first number average molecular weight ("Mmi") is irradiated, for example by treatment with ionizing radiation (e.g., in the form of gamma radiation, X-ray radiation, ultraviolet (UV) light from 100 to 280 nm, a beam of electrons or other charged particles) to obtain a second material 3, which includes cellulose having a second number average molecular weight ("Mmg") lower than the first number average molecular weight. The second material (or the first and second material) is possible

To be mixed with a microorganism (e.g., bacteria or yeast) that can utilize the second and/or first material to produce product 5.

Since the second material C has a cellulose having a reduced recalcitrance, molecular weight relative to the first material, and, in some cases, reduced crystallinity, the second material is generally more dispersible, swellable and/or soluble in the solution containing the microorganism. These properties make the second material 3 more susceptible to chemical, enzymatic and/or biological action (for example, a microorganism) relative to the first material 2, which can significantly increase the production rate and/or production level of the desired product, such as ethanol. Radiation exposure can also sterilize materials.

In some embodiments, the second number average molecular weight ("Mg") is lower than the first number average molecular weight ("Mmi) by more than about 10 95, such as 15, 20, 25, 30, 35, 40, 50, 60 95 or even more than about 75 95.

Ionizing radiation

Each form of radiation ionizes biomass through specific interactions determined by the energy of the radiation. Heavy charged particles mainly ionize matter through Coulomb scattering; moreover, these interactions generate energetic electrons that can further ionize matter. Alpha particles are identical to the nucleus of a helium atom and are produced by the alpha decay of various radioactive nuclei such as isotopes of bismuth, polonium, astatine, radon, francium, radium, several actinoids such as actinium, thorium, uranium, neptunium, curium, californium, americium and plutonium.

When particles are used, they can be neutral (uncharged), positively charged, or negatively charged. When they are charged, charged particles can carry a positive or negative charge, or multiple charges, such as one, two, three, or even four or more charges. In cases where chain separation is desired, positively charged particles may be desirable, in part due to their acidic nature. When particles are used, the particles may have the mass of the rest electron or a greater mass, such as 500, 1000, 1500, or 2000 or more times the mass of the rest electron. For example, the particles may have a mass of from about 1 atomic unit to about 150 atomic units, such as from about 1 60 atomic units to about 50 atomic units, or from about 1 to about 25,

for example 1, 2, 3, 4, 5, 10, 12 or 15 atomic units. Accelerators used to accelerate particles can be US-electrostatic, US-electrodynamic, EE-linear, linear with magnetic induction or continuous. For example, cyclotron accelerators are available from IVA, Belgium, such as the Kpodoigopbt system, and OS-type accelerators are available from KOI, currently IVA Ipdisigia!, such as the OupatiigopF). Ions and ion accelerators are considered in Ipigodisiogu Mysieag Rpuzis5, Keppeii 5. Kgape, dopp Umieu 4 5opv, Ips. (1988), Kt

RheiIes, RILICA V 6 (1997) 4, 177-206, Spy, MMShat T., "Omegmiemzhm oi SPoReop Veat TNegaru",

Soishtryv-Opio, ISVO-IAEA Meeiipo, 18-20 Magsn 2006, maga, U. ei a!., "APegpayipd-Rpaze-Rosivzei 1H-OTI. og Neamu-Iop Medisa! AsseiIegayug5", Rgoseedipd oi ERAS 2006, Ediprigdy, Zsoyapa, and

Yiepeg, S.M. ei ai., "av oi Te Ziregsopdisiipd ESV Iop botsgse Mepyv", Rgoseedipd5 oi ERAS 2000, Mieppa, Aye!yvipa. As a rule, generators are contained in a storage, for example, made of lead or concrete.

Electrons interact by means of Coulomb scattering and inhibition of radiation caused by changes in the speed of electrons. Electrons can be generated by radioactive nuclei that undergo beta decay, such as isotopes of iodine, cesium, technetium, and iridium. Alternatively, an electron gun using thermionic emission can be used as a source of electrons.

Electromagnetic radiation acts through three processes: photoelectric absorption, Compton scattering, and vapor formation. The predominant effect is determined by the energy of the incident radiation and the atomic number of the material. The sum of the interactions leading to the absorption of radiation in the cellulosic material can be expressed using the mass absorption coefficient (see "Iopilayop Kadiayop" in

PCT/O52007/022719).

Electromagnetic radiation is divided into gamma rays, X-rays, ultraviolet rays, infrared rays, microwaves or radio waves, depending on the wavelength.

For example, gamma radiation can be used to irradiate materials.

Referring to fig. 9 and 10 (enlarged view of area K), the gamma emitter 10 includes sources of gamma radiation 408, such as CO tablets, a work table 14 for holding materials to be irradiated, and a storage device 16, for example, made of a plurality of iron plates, which all are contained in a concrete-shielded chamber (storage) 20, which includes a labyrinthine entrance 22 behind a leaded door 26. The accumulator 16 includes a plurality of channels 30, such as sixteen or more channels, which allow gamma radiation sources to pass through the accumulator near the working table

In the process of work, the sample to be irradiated is placed on the work table.

The irradiator is adapted to deliver the desired dose level and to interface the experimental unit 31 with control equipment. The operator then leaves the protective chamber, passing through the entrance in the form of a labyrinth and through a leaded door. The operator occupies the control panel 32, instructing the computer 33 to bring the radiation sources 12 into the working position using the cylinder 36 connected to the hydraulic pump 40.

Gamma radiation has the advantage of a significant depth of penetration into various sample materials.

Sources of gamma rays include radioactive nuclei such as isotopes of cobalt, calcium, technetium, chromium, gallium, indium, iodine, iron, krypton, samarium, selenium, sodium, thallium, and xenon.

X-ray sources include electron beam collisions with metal targets such as tungsten or molybdenum or alloys, or compact light sources such as the commercially produced Iupseap light sources. Sources of ultraviolet radiation include deuterium or cadmium lamps. Infrared radiation sources include ceramic lamps with a sapphire, zinc, or selenide window. Microwave sources include klystrons, KE-type Ziemip sources, or atomic beam sources using hydrogen, oxygen, or nitrogen gas.

Various other irradiation devices can be used in the methods described herein, including field ionization sources, electrostatic ion separators, field ionization generators, thermionic emission sources, ultrahigh frequency discharge ion sources, recirculating or static accelerators, dynamic linear accelerators, accelerators Van de Graafia and bent tandem accelerators. Such devices are disclosed, for example, in US Patent Application Serial No. 61/073665, the entire contents of which are incorporated herein by reference.

Electron beams

In some embodiments, an electron beam is used as a source of radiation. The electron beam has the advantage of high dose levels (eg 1, 5 or even 10 Mrad per second), high performance, less protective insulation and less isolation equipment. Electrons can also be more efficient at providing chain separation. In addition, electrons having an energy of 4-10 MeV can have a penetration depth of 5 to 30 mm or more, for example 40 mm.

Electron beams can be generated, for example, using electrostatic generators, cascade generators, transformer generators, low-energy scanning accelerators, low-energy linear cathode accelerators, linear accelerators, and pulse accelerators. Electrons may be suitable as a source of ionizing radiation, e.g., for relatively thin stacks of material, e.g., less than about 0.5 in. (1.27 cm), e.g., less than 0.4 in. (1.02 cm), 0.3 in. (0.76 cm), 0.2 in (0.51 cm), or less than 0.1 in (0.25). In some embodiments, the energy of each electron in the electron beam is from about 0.3 MeV to about 2.0 MeV (megaelectron volts), such as from about 0.5 MeV to about 1.5

MeV or from about 0.7 MeV to about 1.25 MeV.

In Fig. 11 presents a basic technological scheme 3000, which includes various stages in the sequence of preliminary processing of raw materials with an electron beam. In the first stage, 3010 dry raw materials are supplied from the feedstock source. As discussed above, the dry feedstock from the feedstock source may be preprocessed prior to delivery to the electron beam devices. For example, if the raw material is derived from plant sources, certain parts of the plant material may be removed prior to harvesting the plant material and/or prior to delivering the plant material using a raw material transport device. Alternatively or additionally, as reflected in optional stage 3020, the biomass feedstock can be subjected to mechanical processing (eg, to reduce the average fiber length in the feedstock) prior to delivery to the electron beam irradiation devices.

In step 3030, the dry raw material is moved to a raw material transport device (eg, a conveyor belt) and distributed along the transverse length of the device for

From the transportation of raw materials, it is approximately uniform in volume. This can be done, for example, manually or by inducing a localized vibrational motion at some point of the device for transporting raw materials before processing by means of electron beam irradiation.

In some embodiments, the mixing system supplies a chemical reagent 3045 to the raw material in an optional process 3040 in which a slurry is formed. The combination of water with processed raw materials in the mixing stage 3040 leads to an aqueous suspension of raw materials that can be transported, for example, through a system of pipes, rather than using, for example, a conveyor belt.

The next stage 3050 is a cycle that includes exposure to the raw material (in dry form or in the form of a suspension) of electron beam irradiation from one or more (for example, M) electron beam irradiation devices. The suspension of feedstock is moved through each of the M "streams" of electron beams at stage 3052. Movement through or between the streams may occur at a constant rate, or there may be a pause during passage through each stream, followed by rapid movement to the next stream. At step 3053, a small portion of the feedstock slurry is exposed to each stream for some predetermined exposure time.

Devices for electron beam irradiation can be purchased commercially from (op Veat

Arriisaiopv, Ioimaip-Ia-Meime, Belgium, or Tiap Sogrogaiop, Diego, SA. A typical electron energy can be 1, 2, 4.5, 7.5 or 10 MeV. A typical power of an electron beam irradiation device can be 1, 5, 10, 20, 50, 100, 250 or 500 kW.

The efficiency of depolymerization of the raw material suspension depends on the used electron energy and the applied dose, while the exposure time depends on the power and dose. Typical doses can be 1, 5, 10, 20, 50, 100 or 200 kGy.

When choosing the optimal power characteristics of the device for electron beam irradiation, operating costs, capital costs, depreciation costs and the area of the device are taken into account. When choosing the optimal levels of exposure dose of electron beam irradiation, energy consumption and issues of ecology, safety and health (E5N) are taken into account. When choosing the optimal electron energy, energy consumption is taken into account; in this case, a lower electron energy may be preferable from the point of view of promoting the depolymerization of any suspension of raw materials (see, for example, Voispago, ei a!., Seimiose (2006) 13:601-610).

To ensure a more efficient depolymerization process, it may be preferable to conduct irradiation with an electron beam with a double pass. For example, a raw material transport device can direct the raw material (in dry form or in slurry form) down and in a reverse direction relative to its original direction of transport. Dual-pass systems can handle thicker stock slurries and can provide more uniform depolymerization through the thickness of the stock slurry.

An electron beam irradiation device can generate either a fixed beam or a scanning beam. A scanning beam with long scan sweep lengths and high scan speeds may be preferred, as this can effectively replace a large fixed beam width. In addition, sweep lengths of 0.5, 1, 2 m or more are available.

One suitable device is described in Example 22.

After transporting part of the suspension of raw materials through M devices for irradiation with an electron beam, in some embodiments, it may be necessary, as in step 3060, to mechanically separate the liquid and solid components of the suspension of raw materials. In these embodiments, residual solid particles are filtered from the liquid portion of the raw material suspension and returned to the slurry preparation stage 3040. The solid portion of the raw material suspension is then moved to the next processing stage 3070 using a raw material transport device. In other embodiments, the raw material is maintained in the form of a suspension for further processing.

A beam of heavy ion particles

Particles heavier than electrons can be used to irradiate carbohydrates or materials that include carbohydrates, such as cellulosic materials, lignocellulosic materials, starchy materials, or mixtures of any of these and other materials described herein. For example, protons, a group of helium nuclei, argon ions, silicon ions, neon ions, carbon ions, phosphorus ions, oxygen ions or nitrogen ions can be used. In some embodiments, particles heavier than electrons can induce higher levels of chain separation. In some cases, positively charged particles can induce higher levels of chain separation than negatively charged particles due to their acidity.

Heavier beams can be generated, for example, using linear accelerators

Zo or cyclotrons. In some embodiments, the energy of each particle in the beam is from about 1.0 MeV/atomic unit to about 6000 MeV/atomic unit, such as from about 3 MeV/atomic unit to about 4800 MeV/atomic unit or from about 10 MeV/atomic unit to about 1000 MeV/atomic unit.

Electromagnetic radiation

In embodiments in which exposure is performed using electromagnetic radiation, the electromagnetic radiation may have an energy per photon (in electron volts), for example, greater than 102 eV, such as greater than 105, 104, 105, 105 eV, or even greater than 107 eV. In some embodiments, the electromagnetic radiation has an energy per photon of 107 to 107 eV, such as 107 to 105 eV. Electromagnetic radiation can have a frequency, for example, greater than 1016 Hz, greater than 1077, 1018, 1019, 1029 Hz, or even greater than 102!

Hz. In some embodiments, the electromagnetic radiation has a frequency between 108 and 1022

Hz, for example from 10"? to 107! Hz.

Doses

In some embodiments, exposure (with any radiation source or combination of sources) is conducted until the material receives a dose of at least 0.5 Mrad, such as at least 1.0 Mrad, at least 2.5 Mrad, at least 5.0 Mrad, or at least 10.0 Mrad. In some embodiments, the irradiation is carried out until the material receives a dose of at least 1.0 to 6.0 Mrad, such as 1.5 to 4.0 Mrad.

In some embodiments, the irradiation is carried out at a dose level of 5.0 to 1500.0 kilorads/hour, for example, from 10.0 to 750.0 kilorads/hour or from 50.0 to 350.0 kilorads/hour.

In some embodiments, the implementation uses two or more sources of radiation, such as two or more sources of ionizing radiation. For example, samples can be treated, in any order, with an electron beam followed by gamma radiation and UV radiation having wavelengths from about 100 nm to about 280 nm. In some embodiments, samples are treated with three sources of ionizing radiation, such as an electron beam, gamma radiation, and energy

UV radiation.

Alternatively, in another example, the fibrous biomass material, which includes cellulosic and/or lignocellulosic material, is irradiated and optionally treated with sound energy, such as ultrasound.

In one example of using radiation as a treatment, half-gallon (1.9 L) juice cartons made of white kraft paperboard having a bulk density of 20 lb/ft3 (0.32 g /cmU). The paperboard can be folded flat and then fed into a sequence of three shredder-shredder systems arranged in series, where the product of the first shredder is fed as input to the second shredder and the product of the second shredder is fed as input material into a third device for grinding.

The resulting fibrous material can be sprayed with water and processed using a granulation press operating at room temperature. Compacted granules can be placed in a glass ampoule, from which the air is pumped out under high vacuum, and then refilled with argon gas. The ampoule is sealed in an argon atmosphere. The granules in the ampoule are irradiated with gamma radiation for about 3 hours at a dose level of about 1

Mrad per hour with the production of irradiated material in which the cellulose has a lower molecular weight than the starting material.

Extinction and controlled functionalization of biomass

After treatment with one or more types of ionizing radiation, such as photon radiation (for example, X-rays and gamma rays), exposure to an electron beam, or particles heavier than electrons that are positively or negatively charged (for example, protons or carbon ions), any of hydrocarbon-containing materials or mixtures described in this document become ionized; that is, they include radicals at levels detectable by an electron paramagnetic resonance spectrometer.

The current limit of radical detection is approximately 1077 spins at room temperature.

After ionization, any biomass material that is ionized can be quenched to reduce the level of radicals in the ionized biomass, for example, such that the radicals are no longer detectable by an electron paramagnetic resonance spectrometer. For example, radicals can be quenched by applying sufficient pressure to the biomass and/or by using a fluid medium that

Zo comes into contact with ionized biomass, such as gas or liquid, which reacts (quench) with radicals.

The use of a gas or liquid to at least aid radical quenching can be used to functionalize the ionized biomass with the desired number and type of functional groups, such as carboxylic acid groups, enol groups, aldehyde groups, nitro groups, nitrile groups, amino groups, alkylamino groups, alkyl groups, chloroalkyl groups, or chlorofluoroalkyl groups. groups In some cases, such quenching can increase the stability of some of the ionized biomass materials. For example, quenching can increase the resistance of biomass to oxidation. Functionalization by quenching can also increase the solubility of any biomass described herein, can increase its thermal stability, which can increase the utilization of the material by various microorganisms. For example, the functional groups that are provided to the biomass material by quenching can act as receptor sites for binding by microorganisms, for example to enhance the hydrolysis of cellulose by various microorganisms.

In Fig. 11A illustrates changing the molecular and/or supramolecular structure of the biomass feedstock by pre-treating the biomass feedstock with ionizing radiation, such as electrons or ions with energy sufficient to ionize the feedstock, to provide a first level of radicals. As shown in Fig. 11A, if the ionized biomass remains in the atmosphere, it oxidizes, for example, to such an extent that carboxylic acid groups are formed by reaction with atmospheric oxygen. In some cases, for some materials, such oxidation is desirable because it can contribute to a further decrease in the molecular weight of the carbohydrate-containing biomass, and oxidizing groups, such as carboxylic acid groups, can in some cases be useful for solubility and utilization by the microorganism. However, since radicals can "live" for some time after exposure, such as more than 1 day, 5 days, 3 days, 3 months, 2 months, or even more than 1 year, the properties of the material can continue to change with time, which in some cases can be unwanted

The detection of radicals in irradiated samples using electron paramagnetic resonance spectroscopy and the life time of radicals in such samples are discussed in Vagoiotsa, ii.,

Rpuzisv ip Medisipe apa Vioiodu, 46 (2001), 461-471 and Vapoyuna eyi ai., Kadiatsop Rgoiesiop roziteigu, Moi. 84, Language 1-4, pp. 293-296 (1999). As shown in fig. 114A, the ionized biomass can be quenched to functionalize and/or stabilize the ionized biomass. At any point in time, such as when the material is "live" (still having a significant amount of reactive intermediates such as radicals), "partially live" or fully quenched, the pretreated biomass can be converted into a product, such as a food .

In some embodiments, quenching includes applying pressure to the biomass, for example, by mechanically deforming the biomass, such as direct mechanical compression of the biomass in one, two, or three dimensions, or by applying pressure to a fluid medium in which the biomass is immersed, such as isostatic pressing. In such cases, deformation of the material itself yields radicals that are often trapped in crystalline domains close enough for the radicals to recombine or react with another group. In some cases, the pressure is applied together with the application of heating, such as an amount of heat sufficient to raise the temperature of the biomass to a level above the melting point or to the softening temperature of a biomass component such as lignin, cellulose, or hemicellulose. Heating can increase the mobility of molecules in the polymer material, which can help quench radicals. When pressure is used for extinguishing, the pressure may exceed 1000 psi. in. (6.9 MPa), for example to exceed approximately 1250 lb/sq. in. (8.6 MPa), 1450 lb/sq. in. (10 MPa), 3625 lb/sq. in. (25.2 MPa), 5075 lb/sq. in. (35 MPa), 7250 lb/sq. in. (50 MPa), 10,000 psi. in. (69 MPa) or even more than 15,000 psi. inch (103.4

MPa).

In some embodiments, quenching includes contacting the biomass with a fluid medium, such as a liquid or gas, such as a gas capable of reacting with radicals, such as acetylene or a mixture of acetylene in nitrogen, ethylene, chlorinated ethylenes or chlorofluoroethylenes, propylene, or mixtures of these gases. In other specific embodiments, quenching includes contacting the biomass with a liquid, such as a liquid soluble in the biomass or at least capable of penetrating the biomass and reacting with radicals such as a diene such as 1,5-cyclooctadiene. In some specific embodiments, quenching includes contacting the biomass with an antioxidant, such as vitamin E. If desired, the biomass feedstock may include an antioxidant dispersed therein, and the quenching may occur as a result of contacting the antioxidant dispersed in the biomass feedstock with radicals. Combinations of these and other extinguishing materials can be used.

Other extinguishing methods are possible. For example, for extinguishing any ionized material,

From what is described in this document, it is possible to use any method of quenching radicals in polymer materials, described in Mygaiodia et al., publication of US patent application No. 2008/0067724 and Mygaiodia et al., US patent Mo 7166650. Moreover, for quenching, any -any quenching agent (described as a "sensitizing agent" in the Mygaiodia references above) and/or any antioxidant described in any of the Mygaiodia references can be used on any ionized biomass material.

Functionalization can be enhanced using heavy charged ions, such as any of the heavier ions described herein. For example, if it is desired to increase oxidation, charged oxygen ions can be used for irradiation. If nitrogen functional groups are desired, nitrogen ions or ions that include nitrogen can be used.

Similarly, if sulfur or phosphorus groups are desired, sulfur or phosphorus ions can be used for irradiation.

In some embodiments, after quenching, any of the quenched ionized materials described herein can be further treated with one or more of radiation exposure, such as ionizing or non-ionizing radiation, sonication, pyrolysis, and oxidation to further alter the molecular and/or supramolecular structures.

Particle beam irradiation in fluid media

In some cases, cellulosic or lignocellulosic materials can be exposed to particle beam irradiation in the presence of one or more additional fluid media (eg, gases and/or liquids). The effect on the material of a beam of particles in the presence of one or more additional fluid media can increase the efficiency of processing.

In some embodiments, the material is irradiated with a beam of particles in the presence of a fluid medium, such as air. Particles accelerated in one or more of the types of accelerators described in this document (or in another type of accelerator) exit the accelerator through an outlet (e.g., a thin membrane such as a metal foil), pass through a volume of space occupied by the fluid environment, and then fall on the material. In addition to the direct processing of the material, some of the particles form additional chemical particles by interaction with the particles of the fluid medium (for example, ions and/or radicals generated by various components of the air, such as ozone and nitrogen oxides). These formed chemical particles can also react with the material and can act as initiators of various reactions of destruction of chemical bonds in the material. For example, any oxidizing agent formed can oxidize the material, which can lead to a decrease in molecular weight.

In certain embodiments, additional fluids can be selectively applied to the path of the particle beam before the beam hits the material. As discussed above, reactions between the particles of the beam and the particles of the supplied fluids can form additional chemical particles that react with the material and can contribute to the functionalization of the material and/or otherwise selectively change certain properties of the material. One or more additional fluids can be directed into the path of the beam, for example, from a downpipe. The direction and flow rate of the supplied fluid(s) can be selected according to the desired power or direction of irradiation to control the overall processing efficiency, including both effects resulting from particle processing and effects resulting from the interaction of dynamically generated particles with of the given fluid medium with the material. In addition to air, illustrative fluids that can be supplied to the ion beam include oxygen, nitrogen, one or more noble gases, one or more halogens, and hydrogen.

Irradiation of biomass materials with low bulk density and cooling of irradiated biomass

In the process of treating biomass materials with ionizing radiation, especially at high dose levels, such as levels greater than 0.15 Mrad per second, for example 0.25, 0.35, 0.5, 0.75 Mrad/s or even more than 1 Mrad/ c, biomass materials can store significant amounts of heat, so that the temperature of biomass materials increases. While elevated temperatures may be preferred in some embodiments, such as when a higher reaction rate is desired, it is preferred to control the heating of the biomass to maintain control over ionizing radiation-initiated chemical reactions such as cross-linking, chain scission, and/or graft copolymerization, for example, to maintain process control. Low bulk density materials, such as materials having a bulk density of less than about 0.4 g/cm3, such as less than about 0.35, 0.25, or less than about 0.15 g/cm3, especially when combined with materials that have thin cross-sections, such as fibers that have small cross-sectional dimensions, are generally easier to cool. In addition, photons and particles can primarily penetrate deeper into or through materials of relatively low bulk density, which may allow processing of larger volumes of materials at higher speeds and may enable applications of photons and particles , which have lower energies, such as 0.25, 0.5, 0.75 or 1.0 MeV, which can reduce shielding requirements for safety. Many of the biomass materials described herein can be processed in one or more of the systems shown in FIG. 118, 11C, 110 and 11E, which are described below. The presented systems allow the application to biomass material with a low bulk density of one or more types of ionizing radiation, such as relativistic electrons or electrons in combination with X-rays, at high dose levels, for example at a level greater than 1.0, 1.5, 2 ,5

Mrad/s or even more than 5.0 Mrad/s, and then allow the biomass to cool before applying radiation exposure for the second, third, fourth, fifth, sixth, seventh, eighth, ninth, even tenth time.

For example, in one method of altering the molecular and/or supramolecular structure of the biomass feedstock, the biomass is pretreated at a first temperature with ionizing radiation, such as photons, electrons, or ions (eg, singly or multiply charged cations or anions), for a sufficient time and/or at a sufficient a dose to raise the temperature of the biomass raw material to a second temperature that exceeds the first temperature. The pretreated biomass is then cooled to a temperature below the second temperature. Finally, if desired, the cooled biomass can be treated one or more times with radiation, such as ionizing radiation. If desired, the biomass can be cooled after and/or during each irradiation treatment.

In some embodiments, the cooling of biomass raw materials is carried out to such an extent that after cooling the biomass has a third temperature below the first temperature.

For example, and as explained in more detail below, the treatment of biomass feedstock with ionizing radiation can be carried out by pneumatically transporting the biomass feedstock into a fluid medium such as a gas such as nitrogen or air. To facilitate molecular weight reduction and/or functionalization of the materials, the gas may be saturated with any swelling agent described herein and/or steam. For example, you can use acidic steam. To facilitate the reduction of molecular weight, water

Zo can be acidified with an organic acid such as formic or acetic acid, or with a mineral acid such as sulfuric or hydrochloric acid.

For example, and as explained in more detail below, treatment of biomass raw materials with ionizing radiation can be carried out as the biomass raw materials fall under the influence of gravity.

This process can effectively reduce the bulk density of the biomass feedstock as it is processed, and it can help cool the feedstock. For example, biomass can be transported from a first belt at a first height above ground level, and then it can enter a second belt at a second level above ground level, which is lower than the first level. For example, in some embodiments, the trailing edge of the first strip and the leading edge of the second strip form a gap. Preferably, ionizing radiation, such as a beam of electrons, protons, or other ions, can be applied to the gap region to prevent damage to the biomass transport system.

In the methods described herein, cooling the biomass may include contacting the biomass with a fluid medium, such as a gas at a temperature below the first or second temperature, such as nitrogen gas at about 77K (-1967C). Even water such as water below nominal room temperature (eg 25 C) can be used.

The biomass feedstock may be treated at the first temperature with ionizing radiation for a sufficient period of time and/or at a sufficient dose, such as from about 1 second to about 10 seconds at a dose of from about 0.5 Mrad/s to about 5 Mrad/s, to increase the temperature of the feedstock biomass to a second temperature higher than the first temperature.

After applying radiation exposure, the biomass can be cooled to the second temperature. The cooled treated biomass is treated with radiation, such as ionizing radiation, and then the treated biomass is brought into contact with a microorganism capable of converting at least a portion, such as at least about 1 wt. 95, biomass into a product.

In some embodiments, the method of changing the molecular and/or supramolecular structure of biomass raw materials does not necessarily include preliminary processing of biomass raw materials by reducing one or more sizes of individual fragments of biomass raw materials and applying

From ionizing radiation, such as photons, electrons or ions, to biomass feedstock. In such embodiments, the biomass feedstock to which the ionizing radiation is applied has a bulk density of less than about 0.35 g/cm3, such as less than about 0.3, 0.25, 0.20 g/cm? or less than about 0.15 g/cm3, during the application of ionizing radiation. In such embodiments, the biomass feedstock may be cooled, and then ionizing radiation may be applied to the cooled biomass. In some preferred embodiments, the biomass feedstock is or includes individual fibers and/or particles having a maximum size of no more than about 0.5 mm, such as no more than about 0.25 mm, no more than about 0.1 mm, no more than about 0.05 mm or no more than about 0.025 mm.

Referring in particular to FIG. 1188 and 11C, which show the acquisition, processing, transportation of biomass and the device for irradiation 1170 (shielding not illustrated in the figures). In operation, a sheet of paper 1173, such as fragments of a bleached sheet of kraft paper, is fed from a roller 1172 and delivered to a device 1174, such as a rotary knife cutter. The sheet 1173 is converted into a fibrous material 1112 and delivered to the fiber loading area 1180 by means of a conveyor 1178. If desired, the fibers of the fibrous material can be separated, for example by sieving, into fractions having different I/O0 ratios. In some embodiments, fibrous material 1112, typically of low bulk density and preferably of thin cross-section, is continuously delivered to zone 1180; and in other embodiments, the fibrous material is delivered in batches.

A fan 1182 in the loop 1184 is located adjacent to the fiber loading area 1180 and is capable of moving a fluid medium, such as air, at a speed and volume sufficient to pneumatically circulate the fibrous material 1112 in the direction indicated by arrow 1188 through the loop 1184.

In some embodiments, the air velocity passing through the loop is sufficient to homogeneously disperse and transport the fibrous material throughout the loop 1184. In some embodiments, the flow rate is greater than 2,500 ft/min (760 m/min), such as 5,000 ft/min ( 1520 m/min), 6000 ft/min (1800 m/min) or more, such as 7500 ft/min (2300 m/min) or 8500 ft/min (2600 m/min).

The loaded fibrous material 1112 passing through the loop passes through the application zone 60 1190, which forms part of the loop 1184. Here, any desired additives described herein are introduced, such as a liquid such as water, such as acidified or alkalized water . During operation in application zone 1190, an additive, such as liquid solution 1196, is introduced into the circulating fibrous material through nozzles 98, 99, and 11100. When liquid is applied, the nozzles produce an atomized spray or aerosol that affects the fibers as the fibers pass near the nozzles. Valve 11102 controls the flow of fluid to the respective nozzles 1198, 1199, and 11100. After the desired amount of additive is added, valve 11102 closes.

In some embodiments, the application zone 1190 is two feet (0.6 m) or longer, such as 125 feet (38 m), 150 feet (46 m), 250 feet (76 m), or greater, such as 500 feet (152 m ). Longer application zones allow for application over a longer period of time as the fibrous material passes through the application zone 1190. In some embodiments, the nozzles are spaced, for example, from about three to about four feet (0.9-1.2 m), along the length of the loop 1184.

As the fibrous material passes through loop 1184 and through the radiating portion 11107 of the loop, which includes horn 11109 for delivering ionizing radiation, the fibrous material is exposed to ionizing radiation (shielding not shown).

As the irradiated fibrous material advances through the loop 1184, it is cooled by gases such as air circulating through the loop at high velocities and surrounded by reactive gases such as ozone and/or nitrogen oxides produced by the action of ionizing radiation on the circulating gases. , such as air. After passing through the irradiating portion 11107, a cooling fluid such as a liquid (eg, water) or a gas such as nitrogen at 77K may be introduced into the loop 1184 to assist in cooling the fibrous material. If desired, this process can be repeated more than once, for example 2, 3, 4, 5, 6, 7, 8, 9, 10 times or more, for example 15 times, to deliver the desired dose to the fibrous material. While the long axis of the horn is shown to be along the direction of flow, in some embodiments, the long axis of the horn is transverse to the direction of flow. In some embodiments, an electron beam is used as the primary source of ionizing radiation, and X-rays are used as a secondary source of ionizing radiation. X-rays can be generated by having a metal target, such as tantalum target 11111, on the inside of loop 1184 so that when electrons reach the target, X-rays are emitted.

After delivery of the desired dose to the fibrous material, the fibrous material can be removed from the loop 1184 through the separator 11112, which is selectively connected to the loop 1184 by the section 11114 and the shut-off valve 11116. When valve 11116 opens, another valve also opens, allowing air to enter loop 1184 , replacing the air exiting separator 11112 .

Referring in particular to FIG. 110, which shows a device for irradiating 11121 fibrous material in a fluidized bed with shielding. Fibrous material in a fluid medium, such as a gas, such as compressed air, is delivered to the shielded protective jacket 11123 through the conduit 11125 and to the fluidized bed shielded portion 11127. Counterflows 11131 of the fluid, such as gas, and crossflows 11133 of the fluid, such as gas that is the same as, or different from, the gas being delivered in the opposite direction, combine to cause turbulence in a portion of the bed. Ionizing radiation is applied in part of the fluidized bed as the fibrous material is transported through part of the bed. For example, as shown, it is possible to use three electron beams from three Kpodoihop devices? 11135, 11136 and 11137. Preferably, each beam can penetrate the fluidized bed to a different depth, and/or each beam can emit electrons with different energies, such as 1, 3, and 5 MeV.

As the irradiated fibrous material passes through the system, it is cooled by gases, such as air, circulating through the system at high velocities, and it is surrounded by reactive gases, such as ozone and/or nitrogen oxides, produced by the action of ionizing radiation on the circulating gases. , such as air. If desired, the process can be repeated as many times as desired until the fibrous material receives the desired dose. Although the fluidized bed is illustrated with its long axis horizontal to the ground, in other embodiments the long axis is perpendicular to the ground so that the fibrous material is subject to gravity.

Referring in particular to FIG. 11E, which shows another method of transporting the fibrous material and the device for irradiation 11140 without shielding. The fibrous material 11144 is delivered from the basket 11142 to the first conveyor 11150 at the first level above the ground, and then the material is transferred to the second conveyor 11152 at a lower height than the first conveyor. The rear edge 11160 of the first conveyor and the front edge 11161 of the second conveyor 11152 define a gap of a distance 5. For example, the distance 5 can be from 4 inches (0.1 m) to about 24 inches (0.6 m). The material 11144 has sufficient momentum to free fall under gravity and then enter the second conveyor 11152 without falling into the gap. In the process of free fall, ionizing radiation is applied to the material. This arrangement can be advantageous in that ionizing radiation is less likely to damage the transport system due to the lack of direct contact with radiation exposure.

After passing through the irradiating section, a cooling fluid such as a liquid (eg water) or a gas such as liquid nitrogen at 77K may be applied to the material to assist in cooling the fibrous material. If desired, this process can be repeated more than once, for example 2, 3, 4, 5, 6, 7, 8, 9, 10 times or more, for example 15 times, to deliver the desired dose to the fibrous material. Although, as shown, the long axis of the horn is transverse to the direction of the material flow, a different arrangement of the beams is possible.

In some embodiments, an electron beam is used as the primary source of ionizing radiation, and X-rays are used as a secondary source of ionizing radiation. X-rays can be generated by having a metal target, such as a tantalum target, on the inside of the loop, so that when the electrons reach the target, X-rays are emitted.

In one example of using radiation as a treatment, half-gallon (1.9 L) juice cartons made of white kraft paperboard having a bulk density of 20 lb/ft3 (0.32 g /cm). The paperboard can be folded flat and then fed into a sequence of three shredder-shredder systems arranged in series, where the product of the first shredder is fed as input to the second shredder and the product of the second shredder is fed as input material into a third device for grinding.

The resulting fibrous material can be sprayed with water and processed using a granulation press operating at room temperature. Compacted granules can be placed in a glass ampoule from which the air is pumped out under high vacuum. Granules in an ampoule

Zo is irradiated with gamma radiation for about 3 hours at a dose level of about 1

Mrad per hour with the production of irradiated material in which the cellulose has a lower molecular weight than the starting material.

Ultrasound treatment

One or more series of sonications can be used to process biomass from a wide variety of different sources to extract nutrients from the feedstock and provide partially degraded organic material that serves as an input stream for further processing stages and/or sequences. Sonication may reduce the recalcitrance, molecular weight, and/or crystallinity of feedstocks such as one or more of any of the biomass materials described herein, such as one or more carbohydrate sources such as cellulosic or lignocellulosic materials or starchy materials.

Referring again to FIG. 8, in one method, the first biomass material 2, which includes cellulose having a first number average molecular weight ("Mm1:"), is dispersed in a medium such as water and sonicated or otherwise cavitated to produce a second biomass material 3, which includes cellulose having a second number average molecular weight ("Mmg") lower than the first number average molecular weight. The second material (or, in certain embodiments, the first and second materials) can be mixed with a microorganism (eg, bacteria or yeast) that can utilize the second and/or first material to produce product 5.

Since the second material C has a cellulose having a reduced molecular weight relative to the first material and, in some cases, also a reduced crystallinity, the second material is generally more dispersible, swellable and/or soluble in the solution containing the microorganism, for example greater than 105 microorganisms/ml. These properties make the second material C more susceptible to chemical, enzymatic and/or microbial attack relative to the first material 2, which can significantly increase the production rate and/or production level of the desired product, such as ethanol. Sonication can also sterilize materials, but should not be used as long as the microorganisms are expected to be alive.

In some embodiments, the second number average molecular weight ("Mmg") is lower than the first number average molecular weight ("Mm:") by more than about 10 95 , such as 15, 20, 25, 30, 35, 40, 50, 60 95 or even more than about 75 95.

In some cases, the second material has cellulose having a crystallinity ("Cg") that is lower than the crystallinity (Si) of the cellulose of the first material. For example, (Cg) may be lower than (7C:) by more than about 10 95, such as 15, 20, 25, 30, 35, 40 95 or even more than about 50 95.

In some embodiments, the initial crystallinity index (before sonication) is from about 40 95 to about 87.5 95, such as from about 50 95 to about 75 95 or from about 60 956 to about 70 9b5, and the crystallinity index after irradiation is from from about 10 95 to about 50 95, such as from about 15 95 to about 4595 or from about 2095 to about 4095. However, in certain embodiments, such as after extensive irradiation, the crystallinity index may be less than 5 95. In some embodiments, the material after irradiation is essentially amorphous.

In some embodiments, the initial number average molecular weight (before sonication) is from about 200,000 to about 3,200,000, such as from about 250,000 to about 1,000,000 or from about 250,000 to about 700,000, and the number average molecular weight after irradiation is from about 50,000 to about 200,000, for example, from about 60,000 to about 150,000 or from about 70,000 to about 125,000. However, in some embodiments, such as after extensive irradiation, the number average molecular weight may be less than about 10,000 or even less than about 5,000.

In some embodiments, the second material may have an oxidation level (702) greater than the oxidation level (7101) of the first material. A higher level of oxidation of a material can contribute to its ability to disperse, swell and/or dissolve, further increasing the susceptibility of materials to chemical, enzymatic or microbial attack. In some embodiments, to increase the level of oxidation of the second material relative to the first material, irradiation is carried out in an oxidizing atmosphere, producing a second material that is more oxidized than the first material. For example, the second material can

Zo has a larger number of hydroxyl groups, aldehyde groups, ketone groups, ester groups or carboxylic acid groups, which can increase its hydrophilicity.

In some embodiments, the sonication medium is an aqueous medium. If desired, the medium may include an oxidizing agent such as a peroxide (eg, hydrogen peroxide), a dispersing agent, and/or a buffer. Examples of dispersing agents include ionic dispersing agents, such as sodium lauryl sulfate, and nonionic dispersing agents, such as poly(ethylene glycol).

In other embodiments, the sonication medium is non-aqueous.

For example, sonication can be carried out in a hydrocarbon such as toluene or heptane, in an ether such as diethyl ether or tetrahydrofuran, or even in a liquefied gas such as argon, xenon or nitrogen.

Without being bound by any particular theory, it is believed that sonication destroys the bonds in the cellulose by creating bubbles in the medium containing the cellulose that grow and then collapse. In the process of bubble collapse, which can take place in less than a nanosecond, the implosive force raises the local temperature in the bubble to about 5100 K (in some cases even higher; see, for example, Zyzwiisk ei ai., Mayighe 434, 52-55) and creates a pressure from a few hundred atmospheres to more than 1000 atmospheres or more. It is these high temperatures and pressure that destroy bonds. Additionally, without being bound by any particular theory, the reduced crystallinity is believed to be a consequence, at least in part, of extremely high cooling rates during bubble collapse, which can exceed approximately 10"K/second.

High cooling rates generally prevent the cellulose from organizing and crystallizing, resulting in materials that have reduced crystallinity.

Ultrasonic systems and ultrasonic chemistry are considered, for example, in ОЙ ей айЙ., US patent Mo 5766764; Kobregi5, US patent Mo 5828156; Mazop, Spetivhigu m'yypy OpPgazoshpa, EiIbemieg, Ohio, (1990); BiviiskK (vdyog), Opgazotspa: yb Spetisai, Rnuzis! apa Viioodisa! eMesib5, MSN, MUvipneiit, (1988); Riise, "Sitepi Tgepavz ip Zopospetivighu" Wow! Bosieiu oi Spetivigu, Satrbgidde, (1992); zZiviisK ei a!., App. wow Maieg. 5 29, 295, (1999); BiviisK ei a!., Mayte 353, 414 (1991); NiPeg heri aI.,, Ripuvz. Wow And hey. 69, 1182 (1992); Vaiteg ei a!., Mayte, 352, 414 (1991); BiviisK ei ai., u). At.

Spent boss, 108, 5641 (1986); Tapdu ei aiI., Set. Sott., 2119 (2000); UUapd eyi ai., Admapseid

Maieg., 12, 1137 (2000); And updats ei ai., u. Journal of Science, 201, 22 (2001); Rekaz of her AI., Snet. Sott., bo 988 (2001); Mikpepko ei aI., Apdem/. Spent Ipieg. Eh. (Oesetrbreg 2001); Zpaiyi eyi ai., U. Rpuz. Speth V

103, 3358 (1999); Amimi ei ai., U. Ateg. Spent Zos. 121, 4196 (1999); and Amimi ei ai., U. Ateg. Spent wasps 122, 4331 (2000).

Systems for ultrasound treatment

In Fig. 12 shows a general system in which a flow of biomass material 1210 is mixed with a flow of water 1212 in a container 1214 to form a process flow 1216. The first pump 1218 removes the process flow 1216 from the container 1214 in the direction of the flow cell 1224.

The ultrasonic transducer 1226 transmits ultrasonic energy to the process stream 1216 as the process stream passes through the flow cell 1224. The second pump 1230 removes the process stream 1216 from the flow cell 1224 towards further processing.

The container 1214 includes a first receiver 1232 and a second receiver 1234, in hydraulic communication with the volume 1236. A conveyor (not shown) delivers a stream of biomass material 1210 into the container 1214 through the first receiver 1232. The water stream 1212 enters the container 1214 through the second receiver 1234. In some embodiments, the water flow 1212 enters the volume 1236 along a tangential line, which provides a swirling flow in the volume 1236. In certain embodiments, the biomass material flow 1210 and the water flow 1212 are fed into the volume 1236 along opposite axes to enhance mixing in the volume.

Valve 1238 controls the flow of water stream 1212 through second receiver 1232 to obtain a desired ratio of biomass material to water (eg, approximately 10 95 cellulosic materials, weight to volume). For example, 2000 tons/day of biomass can be combined with 1 million to 1.5 million gallons/day (3800 to 5700 m3/day), such as 1.25 million gallons/day (4700 m3/day), of water.

The mixing of the biomass material and water in the container 1214 is controlled by the size of the volume 1236 and the flow rates of the biomass and water into the volume. In some embodiments, the volume 1236 is sized to provide a minimum residence time for the biomass and water during mixing.

For example, when 2,000 tons/day of biomass and 1.25 million gallons/day (4,700 m3/day) of water are flowing through the vessel 1214, the volume 1236 may be approximately 32,000 gallons (120 m3) to achieve the minimum agitation residence time that is approximately 15 minutes.

Container 1214 includes mixer 1240 in hydraulic communication with volume 1236. Mixer

ZO 1240 mixes the contents of volume 1236, completely dispersing the biomass in the water of the volume. For example, the mixer 1240 may be a rotating blade located in the container 1214. In some embodiments, the mixer 1240 disperses the biomass in water essentially homogeneously.

In addition, the vessel 1214 includes an outlet 1242 in hydraulic communication with the volume 1236 and the process flow 1216. The mixture of biomass and water in the volume 1236 flows out of the vessel 1214 through the outlet 1242. The outlet 1242 is located near the bottom of the vessel 1214 to provide expulsion under the action gravity of the mixture of biomass and water from the container 1214 into the technological stream 1216.

A first pump 1218 (e.g., any of several vortex impeller pumps manufactured by Ez5so Rytr5 4 Sopigoi5, Go5 Apdeijez, Saiogpia) moves the contents of the process stream 1216 toward the flow cell 1224. In some embodiments, the first pump 1218 agitates the contents of the process stream 1216 so that the mixture of cellulosic material and water is substantially homogeneous at the inlet 1220 of the flow cell 1224. For example, the first pump 1218 agitates the process stream 1216, creating a turbulent flow along the process stream between the first pump and the inlet 1220 of the flow cell 1224.

The flow cell 1224 includes a reactor volume 1244 in hydraulic communication with an inlet 1220 and an outlet 1222. In some embodiments, the reactor volume 1244 is a stainless steel pipe capable of withstanding elevated pressure (eg, 10 bar). Additionally or alternatively, the reactor volume 1244 includes a rectangular cross-section.

In addition, the flow cell 1224 includes a heat exchanger 1246 in thermal contact with at least a portion of the reactor volume 1244. A cooling fluid 1248 (eg, water) flows into the heat exchanger 1246 and absorbs the heat generated during the sonication of the process stream 1216 in the volume reactor 1244. In some embodiments, the flow rate of the cooling fluid 1248 in the heat exchanger 1246 is controlled to maintain an approximately constant temperature in the reactor volume 1244.

Additionally or alternatively, the temperature of the coolant 1248 entering the heat exchanger 1246 is controlled to maintain an approximately constant temperature in the reactor volume 1244. In some embodiments, the temperature of the reactor volume 1244 is maintained at 20 to 50 °C, such as 25, 30, 35, 40 or 45 "C. Additionally or alternatively, heat transferred to the cooling fluid medium 1248 from the reactor volume 1244 may be used in other parts of the entire process.

Transition compartment 1226 provides hydraulic communication between reactor volume 1244 and booster 1250 coupled (eg, mechanically coupled using a flange) to ultrasonic transducer 1226. For example, transition compartment 1226 may include a flange and an O-ring system positioned to create a vacuum-tight the connection between the reactor volume 1244 and the booster 1250. In some embodiments, the ultrasonic transducer 1226 is a high-power ultrasonic transducer manufactured by Niei5speg OKgazopisw oi Tej/, Germany.

In operation, generator 1252 supplies electricity to ultrasonic transducer 1252. Ultrasonic transducer 1226 includes a piezoelectric element that converts electrical energy into sound in the ultrasonic range. In some embodiments, materials are sonicated using sound having a frequency of about 16 kHz to about 110 kHz, such as from about 18 kHz to about 75 kHz or from about 20 kHz to about 40 kHz (for example, sound having a frequency of 20 to 40 kHz).

Then the ultrasonic energy is delivered to the working environment through the booster 1248.

Ultrasonic energy passing through the booster 1248 in the reactor volume 1244 creates a series of compressions and rarefiations of the process stream 1216 with an intensity sufficient to cause cavitation of the process stream 1216. The cavitation disaggregates the cellulosic material dispersed in the process stream 1216. Cavitation also results in the production free radicals in the water of process stream 1216. These free radicals act to further destroy the cellulosic material in process stream 1216.

Typically, an ultrasonic energy ranging from 5 to 4000

MJ/mU, for example 10, 25, 50, 100, 250, 500, 750, 1000, 2000 or 3000 MJ/m3. After exposure to ultrasonic energy in reactor volume 1244, process stream 1216 exits flow cell 1224 through outlet channel 1222. A second pump 1230 moves process stream 1216 for further processing (eg, any of several vortex impeller pumps manufactured by E55so Rytr5 45 Sopigoi5, I o5 Apdeie5, SaiIShogpia).

Although certain embodiments have been described, other embodiments are possible.

As an example, although process flow 1216 has been described as a single flow path, other schemes are possible. For example, in some embodiments, the process flow 1216 includes a plurality of parallel flow paths (eg, with a flow rate of 10 gallons/min. (38 L/min.)). Additionally or alternatively, a plurality of parallel flow paths of process flow 1216 flow in separate flow cells and are sonicated in parallel (eg, using a plurality of 16 kW ultrasonic transducers).

As another example, although one ultrasonic transducer 1226 has been described as being coupled to a flow cell 1224, other schemes are possible. In some embodiments, a plurality of ultrasonic transducers 1226 are located in the flow cell 1224 (eg, ten ultrasonic transducers may be located in the flow cell 1224). In some embodiments, the sound waves generated by the plurality of ultrasonic transducers 1226 are time-adjusted (eg, synchronized to be out of phase with each other) to enhance the cavitation effect on the process stream 1216 .

As another example, although a single flow cell 1224 has been described, other designs are possible. In some embodiments, the second pump 1230 moves the process stream into a second flow cell, where the second booster and ultrasonic transducer further sonicate the process stream 1216 .

As another example, although reactor volume 1244 has been described as a closed volume, in certain embodiments, reactor volume 1244 is open to ambient conditions. In such embodiments, the ultrasonic pretreatment can be performed substantially simultaneously with other pretreatment methods. For example, ultrasonic energy can be applied to the process stream 1216 in the volume of the reactor 1244 simultaneously with the supply of electron beams to the process stream 1216.

As another example, although a streaming method is described, other schemes are possible. In some embodiments, the sonication can be performed in a cyclic manner. For example, a volume can be filled with a 10 95 (weight by volume) mixture of biomass in water and exposed to sound at an intensity of from about 50 W/cm2 to about 600 W/cm-, for example from about 75

W/cmg to about 300 W/cmg or from about 95 W/cm2 to about 200 W/cm-.

Additionally or alternatively, the bulk mixture may be sonicated for from about 1 hour to about 24 hours, such as from about 1.5 hours to about

12 hours or from about 2 hours to about 10 hours. In certain embodiments, the material is sonicated for a predetermined period of time and then allowed to stand for a second predetermined period of time before being sonicated again.

Referring further to FIG. 13, in some embodiments, two electroacoustic transducers are mechanically coupled to one horn. As shown, a pair of piezoelectric transducers 60 and 62 are connected to the slotted rectangular horn 64 by respective intermediate connecting horns 70 and 72, the latter of which are also known as auxiliary horns.

The mechanical vibration provided by the transducers responding to the high-frequency electrical energy applied to them is transmitted to the corresponding coupling horns, which can be designed to provide mechanical amplification, for example, with a ratio of 1 to 1.2. Horns are provided with appropriate mounting flanges 74 and 76 to support the transducer and horn system in a stationary housing.

The vibration transmitted from the transducers through the connecting or auxiliary horns is transmitted to the input surface 78 of the horn and transmitted through the horn to the opposite output surface 80, which is in forced contact with the workpiece (not shown) during operation, to which use vibration.

High-frequency electrical energy provided by the power source 82 is supplied to each of the converters electrically connected in parallel through the compensating transformer 84 and corresponding series-coupled capacitors 86 and 90, with one capacitor connected in series by electrical connection to each of the converters. Compensation transformer is also known as "Baishp", which means "balancing device". The compensating transformer includes a magnetic core 92 and a pair of identical coils 94 and 96, also called the primary coil and the secondary coil, respectively.

In some embodiments, the transducers include commercially available piezoelectric transducers such as Model 105 or 502 Vgapbop ShNgasopish Sogrogayop, each designed to operate at 20 kHz and a maximum power of 3 kW. The potential difference when turning on the power to ensure the maximum dynamic amplitude on the output surface of the converter is 930 rms volts. Electric

So the current through the converter can vary between zero and 3.5 amps, depending on the load resistance. At 930 rms volts, the output swing is about 20 micrometers. The maximum deviation of the final voltage for the same dynamic amplitude can therefore be 186 volts. Such a deviation in voltage can lead to large stray currents between the converters. Balancing device 430 provides a balanced condition by providing equal current through the converters, thus eliminating the possibility of stray currents. The wire size in the coil must be selected for the maximum load current specified above, and the maximum potential difference occurring at the input of the coil is 93 volts.

As an alternative to the application of ultrasonic energy, high-frequency rotor-stator devices can be used. This type of device generates high shear microcavitation forces that can disintegrate biomass upon contact with such forces. Two types of commercially available high-frequency rotor-stator dispersing devices are the BZirgaiiop'M devices, which are manufactured by Kgyrr Ipaizigiyeesppik StrnN and supplied to the Roit-OiYiimeg UOecizspiapa ZtrN oi Soppesiisci market, and the Oizrah"M devices, which are manufactured and supplied to the Ika-UMogke market , Ipswich, Ohio, USA The operation of such a microcavitation device is described in US Pat. No. 5,370,999.

Although the ultrasonic transducer 1226 is described as including one or more piezoelectric active elements for generating ultrasonic energy, other schemes are possible. In some embodiments, the ultrasonic transducer 1226 includes active elements made of other types of magnetostrictive material (eg, ferrous metals).

The design and operation of such a high-power ultrasonic transducer is considered in

Napzep et al., US patent Mo 6624539. In some embodiments, ultrasonic energy is transferred to the process flow 16 using an electrohydraulic system.

Although the ultrasonic transducer 1226 is described as utilizing the electromagnetic response of magnetostrictive materials to produce ultrasonic energy, other schemes are possible. In some embodiments, acoustic energy in the form of an intense shock wave using an underwater discharge can be applied to the process flow 16. In some embodiments, ultrasonic energy is transferred to the process stream 16 through a thermodynamic system. For example, high energy density acoustic waves may be generated by applying a potential across an enclosed volume of electrolyte, thereby heating the enclosed volume and causing a pressure increase that is subsequently transmitted through the sound propagating medium (eg, process flow 1216). The scheme and operation of such a thermo-hydraulic converter is considered in Nagitapp ei AI., US patent No. 6383152.

Pyrolysis

One or more pyrolysis processing sequences can be used to process biomass from a plurality of different sources to extract useful substances from the biomass and to obtain partially degraded organic material that serves as an input stream for further stages and/or processing sequences.

Referring again to FIG. 8, in one method, the first biomass material 2, which includes cellulose having a first number average molecular weight ("Mm:1), is subjected to pyrolysis, for example, by heating the first material in a tube furnace, to produce a second material

C, which includes cellulose having a second number average molecular weight ("Mg") that is lower than the first number average molecular weight. The second material (or in certain embodiments, the first and second materials) is mixed with a microorganism (eg, a bacterium or yeast) that can dispose of the second and/or first material, producing product 5.

Since the second biomass material has cellulose having a reduced molecular weight relative to the first material and, in some cases, also reduced crystallinity, the second material is generally more dispersible, swellable and/or soluble in the solution containing the microorganism, e.g. greater than 105 microorganisms/ml. These properties make the second material C more susceptible to chemical, enzymatic and/or microbial attack relative to the first material 2, which can significantly increase the production rate and/or production level of the desired product, such as ethanol. Pyrolysis can also sterilize the first and second materials.

In some embodiments, the second number average molecular weight ("Mmg") is lower than the first number average molecular weight ("Mm:") by more than about 10 95 , such as 15, 20, 25, 30, 35, 40, 50, 60 95 or even more than about 75 95.

In some cases, the second material has cellulose having a crystallinity ("Cg") that is lower than the crystallinity (Si) of the cellulose of the first material. For example, (Cg) may be lower than (7C:) by more than about 10 95, for example 15, 20, 25, 30, 35, 40 95 or

From even more than about 50 95.

In some embodiments, the initial crystallinity index (before pyrolysis) is from about 40 95 to about 87.5 95, such as from about 50 95 to about 75 95 or from about 60 95 to about 70 95, and the crystallinity index after irradiation is from about 10 95 to about 50 95, such as from about 15 95 to about 45 95 or from about 20 956 to about 40 95. However, in certain embodiments, such as after extensive pyrolysis, the crystallinity index may be less than 5 95. In some embodiments implementation of the material after pyrolysis is essentially amorphous.

In some embodiments, the starting number average molecular weight (before pyrolysis) is from about 200,000 to about 3,200,000, such as from about 250,000 to about 1,000,000 or from about 250,000 to about 700,000, and the number average molecular weight after pyrolysis is from about 50,000 to about 200,000, e.g. from about 60,000 to about 150,000 or from about 70,000 to about 125,000. However, in some embodiments, such as after extensive pyrolysis, the number average molecular weight may be less than about 10,000 or even less than about 5,000.

In some embodiments, the second material may have an oxidation level (702) greater than the oxidation level (7101) of the first material. A higher level of oxidation of a material can contribute to its ability to disperse, swell and/or dissolve, further increasing the susceptibility of materials to chemical, enzymatic or microbial attack. In some embodiments, to increase the level of oxidation of the second material relative to the first material, pyrolysis is carried out in an oxidizing atmosphere, producing a second material that is more oxidized than the first material. For example, the second material may have more hydroxyl groups, aldehyde groups, ketone groups, ester groups, or carboxylic acid groups that may increase its hydrophilicity.

In some embodiments, the pyrolysis of materials is continuous. In other embodiments, the material is pyrolyzed for a predetermined period of time and then allowed to cool for a second predetermined period of time before pyrolysis again.

Systems for pyrolysis

In Fig. 14 presents a flow chart 6000, which includes various stages of a pyrolytic system for pre-treatment of raw materials. In the first stage 6010, dry raw material is supplied from the source of raw material.

As described above, the dry biomass from the feedstock source may be preprocessed prior to delivery to the pyrolysis chamber. For example, if the biomass is derived from plant sources, certain portions of the plant material may be removed prior to harvesting the plant material and/or prior to delivering the plant material to the feedstock transport device. Alternatively or additionally, the biomass feedstock may be subjected to mechanical processing 6020 (eg, to reduce the average length of fibers in the feedstock) prior to delivery to the pyrolysis chamber.

After mechanical processing, the biomass undergoes a moisture correction stage 6030. The nature of the moisture correction stage depends on the moisture content of the mechanically processed biomass. As a rule, biomass pyrolysis occurs most efficiently when the moisture content of the raw material is from about 10 95 to about 30 95 (eg, from 15 to 25 95) by weight of the raw material.

If the moisture content of the raw material exceeds approximately 40 wt.95, the excessive heat load provided by the water content of the biomass increases the energy consumption in the subsequent stages of pyrolysis.

In some embodiments, if the biomass has a moisture content greater than 30 wt. 95, drier biomass material 6220, which has a low moisture content, can be mixed with it, producing a mixed feedstock at stage 6030 with an average moisture content that is within the limits indicated above. In certain embodiments, biomass with a high moisture content can simply be dried by distributing the biomass material on a moving conveyor that cycles the biomass through an in-line heating element. The heating element evaporates part of the water present in the raw material.

In some embodiments, if the biomass from stage 6020 has a moisture content that is very low (eg, lower than about 10 wt. 95 ), the mechanically processed biomass can be combined with a wetter feedstock material 6230 , with a higher moisture content, such as sewage sludge. Alternatively or additionally, water 6240 can be added to the dry biomass of stage 6020 to increase the moisture content.

At stage 6040, the biomass, now with its moisture content adjusted to be within acceptable limits, can be preheated in an optional preheat stage 6040. Treatment stage 6040 can be used to increase the temperature of the biomass to 75-150"C in preparation for subsequent biomass pyrolysis Depending on the nature of the feedstock and the specific design of the pyrolysis chamber, preheating the biomass can ensure that the heat distribution in the biomass remains more uniform during pyrolysis and can reduce the heat load on the pyrolysis chamber.

The feedstock is then transported to a pyrolysis chamber to undergo pyrolysis in step 6050. In some embodiments, feedstock transport is aided by adding one or more compressed gases to the feedstock stream 6210. The gases create a pressure gradient in the feedstock transport channel, moving the feedstock into the pyrolysis chamber (and even through the pyrolysis chamber). In certain variants, the implementation of the transportation of raw materials occurs mechanically; namely, the raw material is transported to the pyrolysis chamber by a conveying system that includes a conveyor such as a screw conveyor.

It is also possible to add other gases 6210 to the raw material before the pyrolysis chamber. In some embodiments, for example, one or more catalytic gases may be added to the feedstock to aid in the decomposition of the feedstock in the pyrolysis process. In certain embodiments, one or more absorbers can be added to the feedstock to trap volatile materials released during pyrolysis. For example, various sulfur-based compounds such as sulfides may be released during pyrolysis, and an agent such as hydrogen gas may be added to the feedstock to ensure desulfurization of the pyrolysis products. Hydrogen combines with sulfides to form gaseous hydrogen sulfide, which can be removed from pyrolyzed raw materials.

Pyrolysis of the feedstock in the chamber may include heating the feedstock to relatively high temperatures to ensure partial decomposition of the feedstock. Typically, the feedstock is heated to a temperature in the range of 150 to 1100°C. The temperature to which the feedstock is heated depends on a number of factors, including feedstock composition, average feedstock particle size, moisture content, and desired pyrolysis products. For many types of biomass feedstocks use, for example, pyrolysis temperatures from 300 to 550 "C.

The residence time of the raw material in the pyrolysis chamber, as a rule, depends on a number of factors, including the pyrolysis temperature, the composition of the raw material, the average size of the raw material particles, the moisture content and the desired pyrolysis products. In some embodiments, the feed materials are subjected to pyrolysis at a temperature slightly above the decomposition temperature of the material in an inert atmosphere, such as from about 2°C to about 10°C above the decomposition temperature or from about 33°C to about 7°C above the decomposition temperature. In such embodiments In an embodiment, the material is typically held at this temperature for more than 0.5 hour, such as more than 1.0 hour or more than about 2.0 hour.In other embodiments, the material is pyrolyzed at a temperature well above the decomposition temperature of the material in an inert atmosphere, such as from about 75°C to about 175°C above the decomposition temperature or from about 85°C to about 150°C above the decomposition temperature. In such embodiments, the material is typically held at this temperature for less than 0.5 hour, such as less than 20 minutes, less than 10 minutes, less than 5 minutes, or less than 2 minutes In other embodiments, the material are subjected to pyrolysis at an extreme temperature, for example from about 200 "C to about 500 "C above the decomposition temperature of the material in an inert environment or from about 250 "C to about 400 "C above the decomposition temperature. In such embodiments, the material is typically held at this temperature for less than 1 minute, such as less than 30 seconds, less than 15 seconds, less than 10 seconds, less than 5 seconds, less than 1 second, or less than 500 ms. Such implementation options are usually called instant pyrolysis.

In some embodiments, the feedstock in the chamber is heated relatively quickly to the selected pyrolysis temperature. For example, the chamber can be designed to heat the raw material at a rate of 500 to 11,000 C/s. Typical heating rates of biomass-derived raw material are, for example, 500 to 1,000 C/s.

Turbulent flow of the feed material in the pyrolysis chamber is usually preferred because it provides relatively efficient heat transfer of the feed material from the heating subsystem. Turbulent flow can be achieved, for example, by driving the feed material through the chamber using one or more carrier gases 6210 that are injected. As a rule, the carrier gases are relatively inert with respect to the raw material, even at high temperatures in the pyrolysis chamber. Illustrative carrier gases include, for example, nitrogen, argon, methane, carbon monoxide, and carbon dioxide. Alternatively or additionally,

Mechanical transport systems, such as screw conveyors, can transport raw materials and circulate them in the pyrolysis chamber, creating a turbulent flow of raw materials.

In some variants, pyrolysis of raw materials takes place essentially in the absence of oxygen and other reactive gases. Oxygen can be removed from the pyrolysis chamber by periodically purging the chamber with high-pressure nitrogen (eg, nitrogen pressure of 2 bar or more). After purging the chamber, the gas mixture present in the chamber for pyrolysis (for example, in the process of pyrolysis of raw materials) may include less than 4 mol. 95 oxygen (for example, less than 1 mol. 96 oxygen and even less than 0.5 mol. 95 oxygen). The absence of oxygen ensures the absence of ignition of raw materials at elevated pyrolysis temperatures.

In certain embodiments, relatively small amounts of oxygen can be supplied to the raw material and be present in the pyrolysis process. This method is called oxidative pyrolysis. As a rule, oxidative pyrolysis proceeds in several stages of heating. For example, in the first stage of heating, the raw material is heated in the presence of oxygen to ensure partial oxidation of the raw material. This stage uses up the available oxygen in the pyrolysis chamber. Then, in the subsequent stages of heating, the temperature of the raw material is further increased. However, due to the consumption of all oxygen in the chamber, ignition of raw materials does not occur, and pyrolytic decomposition of raw materials occurs without ignition (for example, with the formation of hydrocarbon products). As a rule, the process of heating the raw material in the pyrolysis chamber to initiate decomposition is endothermic. However, during oxidative pyrolysis, the formation of carbon dioxide by the oxidation of raw materials is an exothermic process. The heat released during the formation of carbon dioxide can contribute to the subsequent heating stages during pyrolysis, thereby reducing the heat load provided by the raw material.

In some embodiments, the pyrolysis takes place in an inert environment, for example, when the raw materials are surrounded by argon or gaseous nitrogen. In certain embodiments, pyrolysis may occur in an oxidizing environment, such as air or argon enriched air. In some embodiments, pyrolysis can occur in reducing ambient conditions, for example, when the raw materials are surrounded by gaseous hydrogen. Various chemicals such as oxidizing agents, reducing agents, acids, or bases can be added to the material prior to or during the pyrolysis process to promote pyrolysis. For example, sulfuric acid 60 can be added, or a peroxide (for example, benzoyl peroxide) can be added.

As discussed above, a number of different processing conditions can be used, depending on factors such as feedstock composition and desired pyrolysis products. For example, for a cellulosic feedstock material, relatively mild pyrolysis conditions can be used, including flash pyrolysis temperatures of 375 to 450°C and residence times of less than 1 second. As another example, for an organic solid waste material such as sewage sludge of water, usually use instant pyrolysis temperatures from 500 to 650 "С, with a residence time between 0.5 and 3 seconds. As a rule, many parameters of the pyrolysis process, including residence time, pyrolysis temperature, raw material turbulence, moisture content, composition of raw materials, composition of pyrolysis products, and total composition of gases, can be regulated automatically by a system of regulators and an automated control system.

After the pyrolysis stage 6050, the pyrolysis products pass through the quenching stage 6250 to reduce the temperature of the products before further processing. Typically, the quench stage 6250 includes spraying the pyrolysis products with streams of cooling water 6260. The cooling water also forms a slurry that includes solid undissolved material and various dissolved products. Also, the product stream contains a mixture that includes various gases, including gaseous products, carrier gases, and other types of process gases.

The flow of products is transported through the gas line installed in the line to the gas separator, which performs the gas separation stage 6060, at which gaseous products and other gases are separated from the suspension formed during quenching of pyrolysis products. The separated mixture of gases is optionally sent to the fan 6130, which increases the pressure of the gas by blowing the mixture with air.

The gas mixture may be subjected to a filtration step 6140 in which the gas mixture is passed through one or more filters (eg, activated carbon filters) to remove particulates and other impurities. In the next stage 6150, the filtered gas can be compressed and stored until use. Alternatively, the filtered gas may be subjected to further processing steps 6160. For example, in some embodiments, the filtered gas may be subjected to condensation to separate various gaseous compounds from the gas mixture. Various compounds may include, for example, various hydrocarbon products (eg, alcohols, alkanes, alkenes, alkynes, ethers) formed in the pyrolysis process. In certain embodiments, the filtered gas containing a mixture of hydrocarbon components may be combined with steam gas 6170

Zo (for example, a mixture of water vapor and oxygen) and undergo a cracking process to reduce the molecular weight of hydrocarbon components.

In some embodiments, the pyrolysis chamber includes heat sources that burn hydrocarbon gases such as methane, propane, and/or butane, heating the feedstock. A portion of the separated gases 6270 may be recirculated in the pyrolysis chamber for combustion, generating process heat to support the pyrolysis process. .

In certain embodiments, process heat can be supplied to the pyrolysis chamber, which can be used to increase the temperature of the raw materials. For example, irradiating raw materials with radiation (eg, gamma radiation, electron beam irradiation, or other types of radiation exposure) can heat raw materials to relatively high temperatures. Heated raw materials can be cooled by a heat exchange system that removes some of the excess heat from the irradiated raw materials. The heat exchange system can be adapted to transport part of the thermal energy into the pyrolysis chamber to heat (or preheat) the raw material, thereby reducing the energy consumption of the pyrolysis process.

A suspension containing liquid and solid pyrolysis products may be subjected to an optional dehydration step 6070, in which excess water may be removed from the suspension by processes such as mechanical compression and evaporation. Excess water 6280 may be filtered and then recycled for further use in quenching pyrolysis decomposition products in stage 6250.

The dewatered slurry then passes through a mechanical separation stage 6080, in which the solid product material 6110 is separated from the liquid product material 6090 by means of a series of fine filters with increasing degrees of purification. In step 6100, the liquid product material 6090 may be further condensed (eg, by evaporation) to remove waste water 6190 and purified by methods such as extraction. Extraction may include the addition of one or more organic solvents 6180, for example, to separate products such as oils from products such as alcohols. Suitable organic solvents include, for example, various hydrocarbons and halogenated hydrocarbons. The purified liquid products 6200 can then be subjected to further stages of processing. The waste water 6190 can be filtered if necessary and recirculated for further use in quenching pyrolysis decomposition products in stage 6250.

After separation in step 6080, the solid product material 6110 is optionally subjected to a drying step 6120, which may include evaporation of water. The solid material 6110 can then be stored for further use or it can be subjected to further processing steps, depending on the situation.

The parameters of the pyrolysis process discussed above are illustrative. As a rule, the values of these parameters can vary widely, depending on the nature of the raw materials and desired products.

Moreover, a number of different methods of pyrolysis can be used, including the use of heat sources such as hydrocarbon flames and/or furnaces, infrared lasers, microwave heaters, induction heaters, resistive heaters, and other heating devices and configurations.

A number of different pyrolysis chambers can be used to decompose raw materials. In some embodiments, for example, pyrolysis of the raw material may include heating the material using a resistive heating element, such as a metal hair or metal tape. Heating can occur through direct contact between the resistive heating element and the material.

In certain embodiments, the pyrolysis may include heating the material by induction, for example, using a Curie point pyrolyzer. In some embodiments, the pyrolysis may include heating the material using radiation, such as infrared radiation. Radiation can be generated by a laser, such as an infrared laser.

In certain embodiments, pyrolysis may include heating the material with convective heat. Convective heat can be generated by a flowing stream of heated gas.

The heated gas may be maintained at a temperature less than about 1200 "C, such as less than about 1000 "C, less than 750 "C, less than 600 "C, less than 400 "C, or even less than 300 C. The heated gas may be maintained at a temperature greater than about 250 "6.

Convective heat can be generated by a heated body surrounding the first material, for example, as in a furnace.

In some embodiments, the pyrolysis may include heating the material with steam at a temperature greater than about 250°C.

Zo The variant of the chamber for pyrolysis is presented in Fig. 15. The chamber 6500 includes an insulated chamber wall 6510 with a vent 6600 for exhaust gases, a plurality of furnaces 6520 that generate heat for the pyrolysis process, a transport channel 6530 for transporting raw materials through the chamber 6500, a screw conveyor 6590 for transporting raw materials through the channel 6530 in a turbulent flow and a quenching system 6540, which includes a screw conveyor 6610 for transporting pyrolysis products, water nozzles 6550 for spraying pyrolysis products with cooling water, and a gas separator for separating gaseous products 6580 from a slurry 6570 containing solid and liquid products.

Another embodiment of the chamber for pyrolysis is presented in Fig. 16. The chamber 6700 includes an insulated chamber wall 6710, a feed channel 6720, an inclined inner wall of the chamber 6730, furnaces 6740 that generate heat for the pyrolysis process, a vent 6750 for waste gases, and a gas separator 6760 for separating gaseous products 6770 from liquid and of solid products 6780. Chamber 6700 is adapted to rotate in the direction shown by arrow 6790 to ensure proper mixing and turbulent flow of raw materials in the chamber.

The next embodiment of the chamber for pyrolysis is presented in Fig. 17. Filament pyrolyzer 1712 includes a sample holder 1713 with a resistive heating element 1714 in the form of a wire wound around an open space defined by the sample holder 1713.

Optionally, the heated element may rotate about an axis 1715 (as indicated by arrow 1716 ) to rotate the material, which includes cellulosic material, in the sample holder 1713 .

The space 1718 defined by the enclosure 1719 is maintained at a temperature above room temperature, for example from 200 to 250 "C. In a typical application, a carrier gas, such as an inert gas, or an oxidizing or reducing gas, moves transversely through the sample holder 1713, in while the resistive heating element rotates and heats to the desired temperature, for example 325 "C. After an appropriate period of time, such as 5 to 10 minutes, the pyrolyzed material is removed from the sample holder. The system presented on

Fig. 17, can be scaled and made continuous. For example, instead of a wire, a screw conveyor can be used as a heating element. Material can continuously enter the sample holder, coming into contact with the heated screw, which pyrolyzes the material. At the same time, the screw can push pyrolyzed material out of the sample holder, allowing new non-pyrolyzed material to enter.

Another embodiment of the chamber for pyrolysis is presented in Fig. 18, which shows a Curie point pyrolyzer 1820 that includes a sample chamber 1821 containing a ferromagnetic foil 1822. The sample chamber 1821 is surrounded by a radio frequency coil 1823.

The space 1824 defined by the enclosure 1825 is maintained at a temperature above room temperature, for example 200 to 250 "C. In a typical application, the carrier gas moves in a transverse direction through the sample holder 1821, while the foil 1822 is subjected to inductive heating by the applied radio frequency field for pyrolysis of the material at the desired temperature.

Another embodiment of the chamber for pyrolysis is presented in Fig. 19. Furnace pyrolyzer 130 includes a movable sample holder 131 and a furnace 132. In normal use, the sample is lowered (as indicated by arrow 137) into the heating zone 135 of the furnace 132, while the carrier gas fills the jacket 136 and moves transversely through the sample holder 131. The sample is heated to a desired temperature for a desired period of time to obtain a pyrolyzed product. The pyrolyzed product is removed from the pyrolyzer by lifting the sample holder (as indicated by arrow 134).

In certain embodiments, as shown in FIG. 20, the cellulosic target 140 can be pyrolyzed by treating the target contained in the vacuum chamber 141 with laser radiation, such as radiation having a wavelength of about 225 nm to about 1500 nm. For example, the target can be destroyed at 266 nm using the fourth harmonic wave of the Ma-mZhA laser (5resiga Rpuzis5, ZSK170, Zap dove, Saiii). The optical configuration shown allows the practically monochromatic light 143 generated by the laser 142 to be directed using mirrors 144 and 145 to the target after passing through the lens 146 in the vacuum chamber 141. As a rule, the pressure in the vacuum chamber is maintained at a level of less than 105 mm Hg. In some embodiments, the implementation uses infrared radiation, for example, radiation from a Ma-mZhac laser, which is 1.06 micrometers. In such embodiments, an infrared-sensitive dye can be mixed with the cellulosic material to produce a cellulosic target. Infrared dye can

To increase the heating of the cellulosic material. Laser destruction is described by Viapspei-Rippspeg et al. in US patent Mo 5942649.

Referring to FIG. 21, in some embodiments, the cellulosic material may be flash pyrolyzed by coating a tungsten filament 150, such as a 5 to 25 mm tungsten filament, with the desired cellulosic material while the material is contained in a vacuum chamber 151. To cause pyrolysis, current is passed through the filament , which causes rapid heating of the hair during the desired period of time. As a rule, heating is continued for seconds, and then the hair is allowed to cool. In some embodiments, the heating is carried out several times to ensure the desired degree of pyrolysis.

In certain embodiments, the carbohydrate-containing biomass material may be heated in the absence of oxygen in a fluidized bed reactor. If desired, the carbohydrate-containing biomass may have relatively thin cross-sections and may include any of the fibrous materials described herein for efficient heat transfer. The material can be heated by heat transfer from hot metal or ceramic, such as glass, pellets or sand in a reactor, and the resulting pyrolyzed liquid or oil can be transported to a central production facility for product manufacture.

Oxidation

One or more oxidative processing sequences can be used to process raw materials from multiple different sources in order to extract useful substances from the raw materials and obtain partially degraded organic material that serves as an input stream for further stages and/or processing sequences.

Referring again to FIG. 8, in one method, the first biomass material 2, which includes cellulose having a first number average molecular weight ("Mmi") and having a first oxygen content (0), is subjected to oxidation, for example, by heating the first material in a tubular furnace in a stream of air or enriched with oxygen air, to obtain a second material 3, which includes cellulose having a second number average molecular weight ("Mkig") and having a second oxygen content (O:) lower than the first oxygen content (701). The second material (or, in certain embodiments, the first and second materials) can, for example, be mixed with a material such as a microorganism to produce a composite 4 or other product 5. Providing a higher level of oxidation 60 can increase the dispersibility of the oxidized material, e.g. , in the solvent.

Such materials can also be combined with a solid and/or liquid. For example, a liquid can be in the form of a solution, and a solid can be in the form of particles. The liquid and/or solid may include a microorganism, such as a bacterium and/or an enzyme.

For example, a bacterium and/or an enzyme can act on a cellulosic or lignocellulosic material to produce a product such as a protein. Illustrative products are described in RIVKOOZ MATEKIAI 5 AMO

SOMROZBITE5, О55М 11/453951, filed on June 15, 2006.

In some embodiments, the second number average molecular weight is no more than 97 96 below the first number average molecular weight, such as no more than 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 30 , 20, 12.5, 10.0, 7.5, 5.0, 4.0, 3.0, 2.5, 2.0 95 or not more than 1.095 below the first number average molecular weight. The amount of molecular weight reduction depends on the application.

For example, in some embodiments, the starting number average molecular weight (before oxidation) is from about 200,000 to about 3,200,000, such as from about 250,000 to about 1,000,000 or from about 250,000 to about 700,000, and the number average molecular weight after pyrolysis is from about 175,000 to about 3,000,000 , such as from about 200,000 to about 750,000 or from about 225,000 to about 600,000.

The resins used can be thermosetting plastics or thermoplastic plastics. Examples of thermoplastic resins include rigid and elastomeric thermoplastics. Non-elastic thermoplastic plastics include polyolefins (eg, polyethylene, polypropylene, or polyolefin copolymers), polyesters (eg, polyethylene terephthalate), polyamides (eg, nylon 6, 6/12, or 6/10), and polyethyleneimines.

Examples of elastomeric thermoplastic resins include elastomeric styrene copolymers (eg, styrene-ethylene-butylene-styrene copolymers), polyamide elastomers (eg, polyester-polyamide copolymers), and ethylene-vinyl acetate copolymers.

In specific embodiments, lignin is used, such as any lignin that is produced by any of the methods described herein.

In some embodiments, the thermoplastic resin has a melt flow rate of 10 g/10 minutes to 60 g/10 minutes, such as 20 g/10 minutes to 50 g/10 minutes or 30 g/10 minutes to 45 g/10 minutes, when measured using AZTM 1238. In certain variants of implementation, it is possible to use compatible mixtures of any of the above thermoplastic resins.

In some embodiments, the thermoplastic resin has a polydispersity index (PDI), such as a ratio of weight average molecular weight to number average molecular weight, greater than 1.5, such as greater than 2.0, greater than 2.5, greater than 5.0, greater than 7.5, or even more than 10.0.

In specific embodiments, polyolefins or mixtures of polyolefins are used as a thermoplastic resin.

Examples of thermosetting resins include natural rubber, butadiene rubber, and polyurethanes.

In some embodiments, the starting number average molecular weight (before oxidation) is from about 200,000 to about 3,200,000, such as from about 250,000 to about 1,000,000 or from about 250,000 to about 700,000, and the number average molecular weight after pyrolysis is from about 50,000 to about 200,000, e.g. from about 60,000 to about 150,000 or from about 70,000 to about 125,000. However, in some embodiments, such as after extensive oxidation, the number average molecular weight may be less than about 10,000 or even less than about 5,000.

In some embodiments, the second oxygen content is at least about five percent greater than the first oxygen content, such as greater than 7.5, 10.0, 12.5, 15.0, or 17.5 95. In some preferred embodiments, the second oxygen content of oxygen is at least about 20.0 956 greater than the oxygen content of the first material. The oxygen content is measured by elemental analysis by pyrolysis of the sample in a furnace operating at 1300 "C or more. A suitable device for elemental analysis is the 1st ESO analyzer SNM5-932 3 furnace for high-temperature pyrolysis MTE-900.

In some embodiments, the oxidation of the first material 200 does not significantly change the crystallinity of the cellulose. However, in some cases, for example after excessive oxidation, the second material has cellulose having a crystallinity (Cg) lower than the crystallinity (101) of the cellulose of the first material. For example, (b2) may be lower than (7Ci) by about 60 more than 5 95, such as 10, 15, 20 95, or even 25 95. This may be desirable to increase the solubility of the materials in a liquid, such as a liquid that includes bacteria and/or enzyme.

In some embodiments, the initial crystallinity index (before oxidation) is from about 40 95 to about 87.5 95, such as from about 50 95 to about 75 95 or from about 60 95 to about 70 95, and the crystallinity index after oxidation is from about 30 95 to about 75.0 95, such as from about 35.0 95 to about 70.0 95 or from about 37.5 956 to about 65.0 95. However, in certain embodiments, such as after extensive oxidation, the crystallinity index may be less than 5 95. In some embodiments, the material is essentially amorphous after oxidation.

Without being bound by any particular theory, it is believed that oxidation increases the number of hydrogen-bonding groups on the cellulose, such as hydroxyl groups, aldehyde groups, ketone groups, carboxylic acid groups, or anhydride groups, which can increase its dispersibility and/or its solubility (for example, in liquid). To further enhance the dispersibility in the resin, the resin may include a component that includes hydrogen-bonding groups, such as one or more anhydride groups, carboxylic acid groups, hydroxyl groups, amide groups, amino groups, or mixtures of any of these groups. In some preferred embodiments, the component includes a polymer that is copolymerized with maleic anhydride and/or has grafted maleic anhydride. Such materials are available from Europe under the trade name EOBAVOMOF).

As a rule, the oxidation of the first material 200 occurs in an oxidizing environment.

For example, oxidation may be achieved by pyrolysis in an oxidizing medium, or may be assisted by pyrolysis in an oxidizing medium such as air or argon enriched air. Various chemicals, such as oxidizing agents, acids, or bases, can be added to the material before or during oxidation to promote oxidation.

For example, a peroxide (for example, benzoyl peroxide) can be added before oxidation.

Systems for oxidation

In Fig. 22 presents a technological scheme 5000, which includes various stages in the system of oxidative pretreatment of raw materials. In the first stage 5010, dry raw material is supplied from the raw material source. The source material may include, for example,

A platform or storage container that is connected to the combined oxidation reactor via a conveyor belt or other device for transporting raw materials.

As described above, the dry feedstock from the feedstock source may be pretreated prior to delivery to the oxidation reactor. For example, if the raw material is derived from plant sources, certain parts of the plant material may be removed before harvesting the plant material and/or before delivering the plant material to the raw material transport device. Alternatively or additionally, the biomass feedstock can be subjected to mechanical processing (for example, to reduce the average fiber length in the feedstock) before delivery to the reactor for oxidation.

After mechanical processing 5020, the raw material 5030 is transported to a mixing system, which supplies water 5150 to the raw material in the process of mechanical mixing.

The combination of water with the processed raw material in the stage of mixing 5040 leads to an aqueous suspension 5050 of the raw material, which can then be treated with one or more oxidizers.

As a rule, one liter of water is added to the mixture for every 0.02 to 1.0 kg of dry raw materials.

The ratio of raw materials and water in the mixture depends on the source of raw materials and specific oxidizers used further in the process in general. For example, in conventional industrial lignocellulosic biomass processing sequences, aqueous suspension 5050 feedstock includes from about 0.5 kg to about 1.0 kg of dry biomass per liter of water.

In some embodiments, also at the stage of mixing the raw material 5040, one or more fiber-protecting additives 5170 may be added to the raw material suspension. Fiber-protecting additives can be used, for example, if the desired product of the processing of lignocellulosic raw materials includes cellulose fibers. illustrative fiber-protecting additives include magnesium compounds such as magnesium hydroxide. Concentrations of fiber-protecting additives in suspension 5050 raw materials can be, for example, from 0.1 to 0.4 95 dry mass of raw biomass.

In certain embodiments, the aqueous suspension 5050 of the raw material can be subjected to optional extraction 5180 with an organic solvent to remove water-insoluble substances from the suspension. For example, extraction of slurry 5050 with one or more organic solvents results in a purified slurry and organic waste stream 5210, which includes water-insoluble materials such as fats, oils, and other nonpolar hydrocarbon-based substances. Suitable solvents for carrying out the extraction of suspension 5050 include, for example, various alcohols, hydrocarbons and halogenated hydrocarbons.

In some embodiments, the aqueous suspension 5050 of the raw material can be subjected to an optional heat treatment 5190 to further prepare the raw material for oxidation.

An example of heat treatment includes heating a suspension of raw materials in the presence of compressed steam. In fibrous biomass raw materials, the compressed steam provides swelling of the fibers, exposing a large part of the fiber surfaces to the aqueous solvent and oxidizing agents, which are applied in the further stages of processing.

In certain embodiments, the aqueous slurry of feedstock 5050 can be optionally treated with basic substances 5200. Treatment with one or more basic substances can facilitate the separation of lignin from cellulose in the lignocellulosic biomass feedstock, thereby improving subsequent oxidation of the feedstock. Illustrative basic substances include hydroxides of alkali and alkaline earth metals, such as sodium hydroxide, potassium hydroxide and calcium hydroxide. Generally, a plurality of basic substances can be used, usually in concentrations of from about 0.01 95 to about 0.5 95 of the dry weight of the raw material.

Aqueous slurry 5050 of feedstock is transported (eg, through an in-line piping system) to a chamber, which may be a pre-oxidative processing chamber or an oxidation reactor. At the stage of preliminary oxidation processing 5060, one or more oxidizing agents 5160 are added to the liquid suspension 5050 to form an oxidizing medium. In some embodiments, for example, oxidizing agents 5160 may include hydrogen peroxide. Hydrogen peroxide can be added to suspension 5050 as an aqueous solution and in ratios ranging from 3 to 30-35 wt. 95 suspension 5050. Hydrogen peroxide has a number of advantages as an oxidizer. For example, an aqueous solution of hydrogen peroxide is relatively inexpensive, relatively chemically stable, and not particularly harmful relative to other oxidizing agents (and thus does not require burdensome storage procedures and expensive protective equipment). Moreover, hydrogen peroxide decomposes with the formation of water in the process of oxidation of raw materials, so that the treatment of the waste stream is relatively simple and inexpensive.

In certain embodiments, oxidizing agents 5160 may include oxygen (e.g.,

Co) gaseous oxygen) either alone or in combination with hydrogen peroxide. Oxygen gas can bubble the 5050 slurry in ratios ranging from 0.5 to 10 wt. 95 of slurry 5050. Alternatively or additionally, gaseous oxygen may also be fed into the aqueous phase in equilibrium with slurry 5050 (eg, a steam head above slurry 5050). Oxygen gas can be supplied to either the pre-oxidative processing chamber or the oxidation reactor (or both), depending on the configuration of the oxidative processing system. As a rule, for example, the partial pressure of oxygen in the vapor above the suspension 5050 exceeds the atmospheric pressure of oxygen and is in the range from 0.5 to 35 bar, depending on the nature of the raw material.

Oxygen gas can be supplied in its pure form or it can be mixed with one or more carrier gases. For example, in some embodiments, high-pressure air provides oxygen in the water vapor. In certain embodiments, gaseous oxygen can be continuously supplied in the vapor phase to ensure that the oxygen concentration in the vapor remains within certain predetermined limits during the processing of raw materials. In some embodiments, gaseous oxygen may first be supplied in sufficient concentration to oxidize the feedstock, and then the feedstock may be transported to a closed, pressurized vessel (eg, an oxidation reactor) for processing.

In certain embodiments, the oxidants 5160 may include generated oxygen (eg, oxygen radicals). As a rule, the generated oxygen is produced as needed in the oxidation reactor or in a chamber in hydraulic communication with the oxidation reactor by means of one or more decomposition reactions. For example, in some embodiments, the generated oxygen can be formed in a reaction between MO and O2 in a mixture of gases or in solution. In certain embodiments, the generated oxygen may be produced during the decomposition of the NOSI in solution. Other methods by which the generated oxygen can be produced include electrochemical generation, for example in an electrolyte solution.

As a rule, the oxygen formed is an effective oxidant due to the relatively high reactivity of oxygen radicals. However, the oxygen produced can also be a relatively selective oxidant. For example, when lignocellulosic raw materials are treated with generated oxygen, the selective oxidation of lignin with respect to other components of raw materials, such as cellulose, takes place. As a result of the oxidation of raw materials, the resulting oxygen provides a method of selective removal of the lignin fraction from certain types of raw materials. As a rule, to achieve effective oxidation, concentrations of the generated oxygen from about 0.5 to 5 95 of the dry weight of the raw material are used.

Without regard to theory, it is believed that the generated oxygen reacts with lignocellulosic raw materials by at least two different mechanisms. In the first mechanism, the generated oxygen undergoes a reaction of attachment to lignin, leading to partial oxidation of lignin, which solubilizes lignin in an aqueous solution. As a result, solubilized lignin can be removed from another part of the raw material by washing. In the second mechanism, the formed oxygen destroys butane cross-links and/or opens aromatic rings that are connected through butane cross-links. As a result, the solubility of the aqueous lignin solution increases, and the lignin fraction can be separated from the rest of the raw material by washing.

In some embodiments, the oxidizing agents 5160 include ozone (Oz). The use of ozone can introduce some conditions for the handling of chemical reagents in the sequence of oxidative processing. When heated excessively vigorously, an aqueous solution of ozone can rapidly decompose, with potentially adverse consequences for both the people operating the system and the system equipment. Thus, ozone, as a rule, is formed in a thermally insulated container with thickened walls, separate from the container containing the suspension of raw materials, and transported to it at the appropriate stage of the method.

Without being bound by theory, it is believed that ozone decomposes into oxygen and oxygen radicals, and that oxygen radicals (for example, the oxygen formed) are responsible for the oxidizing properties of ozone as described above. Ozone tends to preferentially oxidize the lignin fraction in lignocellulosic materials, leaving the cellulose fraction relatively unaffected.

The conditions for the oxidation of biomass raw materials based on ozone, as a rule, depend on the nature of the biomass. For example, for cellulosic and/or lignocellulosic raw materials, effective oxidation of raw materials is ensured by ozone concentrations from 0.1 to 20 g/m3 of dry raw materials. As a rule, the water content in the 5050 suspension is from 10 to 80 wt. 95 (for example, from 40 to 60 wt. 95). In the process of ozone-based oxidation, the temperature of the 5050 slurry can be maintained between 0 and 100 °C to avoid intensive ozone decomposition.

In some embodiments, the slurry 5050 feedstock may be treated with an aqueous alkaline solution that includes one or more alkali or alkaline earth metal hydroxides, such as sodium hydroxide, potassium hydroxide, and calcium hydroxide, and may then be treated with an ozone-containing gas in an oxidation reactor. It was found that this process significantly increases the decomposition of biomass in suspension 5050. As a rule, for example, the concentration of hydroxide ions in an alkaline solution is from 0.001 to 10 wt. 95 suspension 5050. After wetting the raw material by contact with an alkaline solution, ozone-containing gas is supplied to the reactor for oxidation, where it contacts the raw material and oxidizes it.

Oxidizers 5160 may also include other substances. In some embodiments, for example, halogen-based oxidants such as chlorine and oxychloride substances (eg, hypochlorite) can be added to slurry 5050. In certain embodiments, nitrogen-containing oxidants can be added to the suspension 5050. Illustrative nitrogen-containing oxidizing agents include, for example, MO and MO." Nitrogen-containing substances in suspension 5050 can also combine with oxygen, creating additional oxidants. For example, both MO and MO' combine with oxygen in the 5050 slurry to form nitrate compounds that are effective oxidizing agents for biomass feedstocks. Halogen- and nitrogen-based oxidants can, in some embodiments, provide bleaching of the biomass feedstock, depending on the nature of the feedstock. Bleaching may be desirable for certain biomass-derived products that are extracted in further processing stages.

Other oxidizing agents may include, for example, various peroxyacids, peroxyacetic acids, persulfates, percarbonates, permanganates, osmium tetroxide, and chromium oxides.

After the stage of preliminary oxidation processing 5060, the slurry 5050 of the raw material is oxidized in the stage 5070. If the oxidants 5160 are added to the slurry 5050 in the reactor for oxidation, then the oxidation proceeds in the same reactor. Alternatively, if oxidizers 5160 are added to the slurry 5050 in the pretreatment chamber, then the slurry 5050 is transported to the reactor for oxidation through a connecting pipe system. After entering the oxidation reactor, the oxidation of the biomass feedstock continues under a controlled set of ambient conditions. As a rule, for example, an oxidation reactor is a cylindrical container that is closed from the external environment and is under pressure. Both cyclic and continuous operation are possible, although environmental conditions are generally easier to control in flow-through cyclic processing operations.

Oxidation of the suspension of 5050 raw materials, as a rule, takes place in the reactor for oxidation at elevated temperatures. For example, the temperature of the 5050 slurry in the oxidation reactor is generally maintained above 100 "C, in the range from 120 to 240 "C. For many types of biomass feedstocks, oxidation is particularly effective if the temperature of the slurry 5050 is maintained between 150 and 220°C. The slurry 5050 can be heated using various heat transfer devices. For example, in some embodiments, the oxidation reactor contacts a heating bath that includes oil or molten salts. In certain embodiments, a series of heat exchange tubes surround and contact the oxidation reactor, and circulation of hot fluid in the tubes heats the slurry 5050 in the reactor. Other heating devices that may be used to heat the slurry 5050 include, for example, resistive heating elements, induction heaters and microwave sources.

The residence time of the feedstock suspension 5050 in the feedstock oxidation reactor can be varied if desired. As a rule, the 5050 suspension is in the reactor during oxidation from 1 minute to 60 minutes. For a relatively soft biomass material such as lignocellulosic material, the residence time in the reactor for oxidation can be from 5 minutes to 30 minutes, for example, at a reactor oxygen pressure of 3 to 12 bar and a slurry temperature of 160 to 210 °C . However, for other types of feedstock, the residence time in the oxidation reactor can be longer, for example up to 48 hours. To determine the appropriate residence time of the 5050 slurry in the oxidation reactor, aliquots of the slurry can be withdrawn from the reactor at certain time intervals and analyzed to determine the concentrations of specific products , of interest, such as complex saccharides Information on the increase in concentrations of certain products in suspension 5050 as a function of time can be used to determine residence times for specific classes of feedstock material.

In some embodiments, in the process of oxidation of the suspension of 5050 raw materials, it is possible to correct the pH of the suspension by adding one or more chemicals to the oxidation reactor. For example, in certain embodiments, oxidation occurs more efficiently in the pH range of approximately 9-11. To maintain the pH in this range, substances such as hydroxides of alkali and alkaline earth metals, carbonates, ammonia and buffer solutions can be fed into the reactor for oxidation.

Circulation of the slurry 5050 may be important in the oxidation process to ensure sufficient contact between the oxidizers 5160 and the feedstock. Circulation of suspension can be achieved

From using different methods. For example, in some embodiments, the oxidation reactor may be equipped with a mechanical stirring device that includes impeller blades or a bladed wheel. In certain embodiments, the oxidation reactor may be a loop reactor in which the aqueous solvent in which the feedstock is suspended is simultaneously drained from the bottom of the reactor and recirculated to the top of the reactor by pumping, thereby ensuring that the slurry is continuously stirred many times and does not stagnate in the reactor.

After the oxidation of the raw material is complete, the slurry is transported to the separation device, where the mechanical separation stage 5080 proceeds. Typically, the mechanical separation stage 5080 includes one or more fine filtration stages with an increasing degree of slurry purification for mechanical separation of solid and liquid components.

The liquid phase 5090 is separated from the solid phase 5100 and the two phases are then processed independently. The solid phase 5100 may optionally be subjected to a drying step 5120, for example, in a drying device. The drying step 5120 may include, for example, mechanically distributing the solid material on the surface to dry and evaporate water from the solid phase 5100 by gently heating the solid material.

After the drying step 5120 (or, alternatively, without the drying step 5120 ), the solid phase 5100 is transported for further processing steps 5140 .

The liquid phase 5090 may optionally be subjected to a drying step 5110 to reduce the concentration of water in the liquid phase. In some embodiments, for example, the drying step 5110 may include evaporation and/or distillation, and/or extraction of water from the liquid phase 5090 by gently heating the liquid. Alternatively or additionally, one or more chemical dewatering reagents may be used to remove water from the liquid phase of 5090.

After the drying step 5110 (or, alternatively, without the drying step 5110), the liquid phase 5090 is transported for further processing steps 5130, which may include various chemical and biological processing steps, such as chemical and/or enzymatic hydrolysis.

The drying stage 5110 produces a waste stream 5220, an aqueous solution that includes dissolved chemicals such as acids and bases in relatively low concentrations. Treatment of the waste stream 5220 may include, for example, pH neutralization with one or more mineral acids or bases. Depending on the concentration of dissolved salts in the waste stream 60 5220, the solution may undergo partial deionization (for example, by passing the waste stream through an ion exchange system). The waste stream, which includes primarily water, may then be recycled into the overall process (eg, as water 5150), may be diverted to another process, or may be discarded.

Typically, in the case of lignocellulosic biomass feedstock, after the separation step 5070, the liquid phase 5090 includes a variety of soluble poly- and oligosaccharides, which can then be separated and/or reduced to shorter chain length saccharides by further processing steps.

The solid phase 5100, as a rule, includes mainly cellulose, for example, with smaller amounts of hemicellulose and lignin products.

In some embodiments, the oxidation can be carried out at an elevated temperature in a reactor, such as a pyrolysis chamber. For example, referring again to FIG. 17, the feed materials may be oxidized in a filament pyrolyzer 1712. In a typical application, an oxidizing carrier gas, such as air or an air/argon mixture, is moved transversely through the sample holder 1713 while the resistive heating element is rotated and heated to a desired temperature, eg 325 °C. After an appropriate period of time, eg 5 to 10 minutes, the oxidized material is removed from the sample holder.

The system presented in Fig. 2, can be scaled and made continuous. For example, instead of a wire, a screw conveyor can be used as a heating element.

Material can continuously enter the sample holder, impinging on the heated screw, which pyrolyzes the material. At the same time, the screw can push oxidized material out of the sample holder, allowing new, unoxidized material to enter.

Referring again to FIG. 18, the raw material can be oxidized in a pyrolyzer point by point

Curie 1820. In a typical application, an oxidizing carrier gas moves transversely through the sample chamber 1821 while the foil 1822 is inductively heated by the radio frequency field applied to oxidize the material at the desired temperature.

Referring again to FIG. 19, the feed material may be oxidized in the pyrolyzer furnace 130. In a typical application, the sample is lowered (as indicated by arrow 137) into the heating zone 135 of the furnace 132, while the carrier gas fills the jacket 136 and moves transversely through the sample holder 131. The sample heated to the desired temperature for the desired time

From the period of time for obtaining the pyrolyzed product. The pyrolyzed product is removed from the pyrolyzer by lifting the sample holder (as indicated by arrow 134).

Referring again to FIG. 20, the raw material may be oxidized by forming a cellulosic target 140 together with an oxidizer such as peroxide and treating the target contained in a vacuum chamber 141 with laser radiation, such as radiation having a wavelength of about 225 nm to about 1600 nm. The optical configuration shown allows the practically monochromatic light 143 generated by the laser 142 to be directed using mirrors 144 and 145 to the target after passing through the lens 146 in the vacuum chamber 141. As a rule, the pressure in the vacuum chamber is maintained at a level of less than 105 mm Hg. In some embodiments, the implementation uses infrared radiation, for example radiation from a Ma-xAs laser, which is 1.06 micrometers. In such embodiments, an infrared-sensitive dye can be mixed with the cellulosic material to produce a cellulosic target. The infrared dye can increase the heating of the cellulosic material. Destruction by a laser is described by Viapspei-Rippspeg ei ai. in US patent Mo 5942649.

Referring again to FIG. 21, raw materials can be rapidly oxidized by coating tungsten filament 150 with an oxidizing agent such as peroxide with a desired cellulosic material while the material is in a vacuum chamber 151. To provide oxidation, a current is passed through the filament causing the filament to rapidly heat up for a desired period of time As a rule, heating is continued for several seconds, and then the hair is allowed to cool. In some embodiments, the heating is carried out several times to ensure the desired degree of oxidation.

Referring again to FIG. 12, in some embodiments, raw materials can be oxidized using sound and/or cavitation. Typically, to achieve oxidation, materials are sonicated in an oxidizing medium, such as water saturated with oxygen or another chemical oxidizer, such as hydrogen peroxide.

Referring again to FIG. 9 and 10, in certain embodiments, ionizing radiation is used to promote oxidation of raw materials. As a rule, to achieve oxidation, materials are irradiated in an oxidizing medium, such as air or oxygen. For example, gamma radiation or electron beam irradiation can be used to irradiate materials.

Other processing methods

Steam explosion can be used alone without any of the methods described in this document or in combination with any of the methods described in this document.

In Fig. 23 shows a general representation of the entire process of converting a fiber source 400 into a product 450, such as ethanol, using a method that includes crushing and steam explosion to obtain a fibrous material 401, which is then hydrolyzed and converted, such as fermented, to produce a product. The fiber source can be converted into fibrous material 401 in a number of possible ways, including at least one crushing method and at least one steam explosion method.

For example, one embodiment involves comminuting the fiber source, followed by an optional screening step(s) and an optional additional comminution step(s) to produce a comminuted fiber source 402, which may then be steam exploded to form a fibrous material 401. After the process steam explosion is optionally followed by a fiber extraction process to remove liquids or "solution" 404 formed in the steam explosion process. The resulting material, after the steam explosion of the shredded fiber source, may be further crushed in an optional additional crushing stage(s) and/or an optional screening stage(s).

In another method, the fibrous material 401 is first steam-exploded to form a steam-exploded fiber source 410. The resulting steam-exploded fiber source is then subjected to an optional fiber extraction process to remove liquids or solution. The resulting steam-exploded fiber source can then be crushed to form a fibrous material. The steam-exploded fiber source may also be subjected to one or more additional sieving steps and/or one or more optional additional crushing steps. The method of crushing and steam explosion of the fiber source to obtain crushed and steam-exploded fibrous material is further discussed below.

Prior to crushing or steam explosion, the fiber source may be cut into fragments or strips of confetti-like material. Crushing processes can occur in the dry state (eg, having less than 0.25 wt. 95 of absorbed water), in the hydrated state, or

Even when the material is partially or completely immersed in a liquid such as water or isopropanol. Also, the method may optionally include stages of drying the output stream after steam explosion or crushing to provide additional stages of dry crushing or steam explosion. The stages of crushing, sieving and steam explosion can be carried out in the presence or absence of various chemical solutions.

In the steam explosion method, the fiber source or the fragmented fiber source is contacted with steam under high pressure, and the steam diffuses into the structures of the fiber source (eg, lignocellulosic structures). The steam is then condensed under high pressure, thereby "wetting" the fiber source. Moisture in the fiber source can hydrolyze any acetyl groups in the fiber source (eg, acetyl groups in hemicellulose fractions) to form organic acids such as acetic and uronic acids. Acids, in turn, can catalyze the depolymerization of hemicellulose, the release of xylan and limited amounts of glucan. Then, when the pressure is released, the "soaked" fiber source (or crushed fiber source, etc.) "explodes". The condensed moisture evaporates instantly due to the sudden pressure drop, and the expansion of the water vapor exerts a shear force on the fiber source (or shredded fiber source, etc.). Sufficient shear force leads to mechanical destruction of the internal structures (for example, lignocellulosic structures) of the fiber source.

The shredded and steam-exploded fibrous material is then converted into a useful product such as ethanol. One way to convert the fibrous material is through hydrolysis to produce fermentable sugars 412, which are then fermented to produce a product. You can also use other known or unknown methods of converting fibrous materials into fuel.

In some embodiments, prior to mixing with the microorganism, the shredded and steam-exploded fibrous material 401 is sterilized to kill any competing microorganisms that may be present on the fibrous material. For example, the fibrous material can be sterilized by exposing the fibrous material to radiation such as infrared radiation, ultraviolet radiation, or ionizing radiation such as gamma radiation. Microorganisms can also be destroyed using chemical sterilants such as bleach (for example, sodium hypochlorite), chlorhexidine, or ethylene oxide.

One of the methods of hydrolysis of shredded and steam-exploded fibrous material is the use of cellulases. Cellulases are a group of enzymes that act synergistically to hydrolyze cellulose. You can also use the commercially available enzyme complex AssayPegazeF 1000, which contains a complex of enzymes that break down lignocellulosic biomass into fermentable sugars.

According to current understanding, cellulase components include endoglucanases, exoglucanases (cellobiohydrolases) and β-glucosidases (cellobiases). Synergism between cellulase components exists when hydrolysis by a combination of two or more components exceeds the sum of the types of activity exhibited by individual components. The generally accepted mechanism of action of the cellulase system (in particular, the system of T. Iopdibgaspiasht) on crystalline cellulose is that endoglucanase hydrolyzes the internal B-1,4-glycosidic bonds of amorphous regions, thereby increasing the number of exposed non-reducing ends. Then, exoglucanases cleave cellobiose elements from non-reducing ends, which, in turn, are hydrolyzed by brglucosidases to individual glucose elements. There are several configurations of both endo- and exoglucanases, distinguished by their stereospecificity. As a rule, optimal hydrolysis of cellulose requires a synergistic action of components in different configurations. However, cellulases are more prone to hydrolyze the amorphous regions of cellulose. There is a linear relationship between crystallinity and hydrolysis rates, with higher crystallinity indices corresponding to slower enzyme hydrolysis rates. Amorphous regions of cellulose are hydrolyzed at twice the rate of crystalline regions. Hydrolysis of shredded and steam-exploded fibrous material can be carried out using any method of biomass hydrolysis.

Steam explosion of biomass sometimes leads to the formation of by-products, such as toxic substances, which are inhibitory to the activity of microbes and enzymes. Thus, the method of converting the shredded and steam-exploded fibrous material into a product may optionally include a liming stage prior to fermentation to precipitate some of the toxic substances. For example, the pH value of the shredded and steam-exploded fibrous material can be raised to above pH 10 by adding calcium hydroxide (Ca(OH)g), followed by a step of lowering the pH to about 5 by adding

From Na5O»h. The calcined fibrous material can then be used as is without removing the precipitate. As shown in Fig. 23, an optional liming stage is carried out immediately before the hydrolysis stage of the shredded and steam-exploded fibrous material, but it is also envisaged to carry out the liming stage after the hydrolysis stage and before the fermentation stage.

In Fig. 24 shows an example of a steam explosion device 460. The steam explosion device 460 includes a reaction chamber 462 into which a fiber source and/or fibrous material is placed through a fiber source inlet 464. The reaction chamber is closed by closing the fiber source inlet valve 465. In addition, the reaction chamber includes a compressed vapor outlet 466, which includes a steam valve 467. In addition, the reaction chamber includes an explosion pressure relief outlet 468, which includes an outlet valve 469, which is connected to the cyclone 470 through a connecting pipe 472 When the reaction chamber includes the fiber source and/or the crushed fiber source and is closed by the shut-off valves 465, 467, and 469, steam is delivered to the reaction chamber 462 by opening the steam inlet valve 467, allowing the steam to escape through the steam inlet 466. After Once the reaction chamber reaches the desired temperature, which can take approximately 20-60 seconds, the dwell time begins. The reaction temperature is maintained at the desired temperature for the desired holding time, which is typically about 10 seconds to 5 minutes. At the end of the holding period, the outlet valve opens, allowing the blast pressure to be released. The depressurization process moves the contents of the reaction chamber 462 from the depressurization outlet 468, through the connecting pipe 472, into the cyclone 470. The steam-exploded fiber source or fibrous material then exits the cyclone in slurry form into a collection basket 474, while most of the residual steam exits the cyclone to the atmosphere through vent 476. In addition, the steam explosion device includes a purge outlet 478 with a purge outlet valve 479 in communication (with connecting pipe 472. Valve 479 outlet for washing is closed during the use of the steam explosion device 460 during steam explosion, but is open during the washing process of the reaction chamber 462. The set temperature of the reaction chamber 462 is preferably from 180 to 240 "C or from 200 to 220 "C. The holding time is preferably is from 10 60 seconds to 30 minutes or from 30 seconds to 10 minutes or from 1 minute to 5 minutes.

Since the steam explosion process results in a deposit in the form of a steam-exploded fibrous material, the steam-exploded fibrous material may not necessarily be subjected to a fiber drawing process where a "solution" is separated from the steam-exploded fibrous material. This fiber extraction step is desirable because it enables further crushing and/or screening processes and can allow the conversion of the fibrous material into a product. The fiber separation process is carried out using a mesh fabric to separate the fibers from the solution. In addition, drying processes can also be included to obtain a fibrous material or a steam-exploded fiber source for further processing.

Any processing method described in this document can be used at pressures above or below normal Earth atmospheric pressure. For example, any process that uses radiation exposure, sonication, oxidation, pyrolysis, steam explosion, or combinations of any of these processes to produce materials that include a hydrocarbon can be conducted under high pressures that can increase reaction rates. For example, any process or combination of processes can be carried out at a pressure greater than about 25 MPa, such as greater than 50, 75, 100 MPa, 150, 200, 250, 350, 500, 750, 1000 MPa, or greater than 1500 MPa.

Combination of irradiation, sonication and/or oxidation devices

In some embodiments, it may be preferable to combine two or more of the individual devices for irradiation, sonication, pyrolysis, or oxidation into a single hybrid device. For such a hybrid device, several processes can be carried out sequentially or even simultaneously, which has the advantage of increasing pretreatment productivity and potential cost savings.

For example, the processes of electron beam irradiation and ultrasound treatment are considered.

Each individual process is effective in reducing the average molecular weight of the cellulosic material by an order of magnitude or more, and several orders of magnitude when performed sequentially.

Both irradiation and sonication processes can be applied using a hybrid electron beam/sonication device such as

Z is illustrated in Fig. 25. A hybrid electron beam/ultrasound treatment device 2500 is shown above an empty bath (3-5 cm deep) for a suspension of cellulosic material 2550 dispersed in an aqueous oxidizing medium such as hydrogen peroxide or carbamide peroxide. The hybrid device 2500 has a power source 2510 that powers both the electron beam emitter 2540 and the sonication horns 2530 .

Electron beam emitter 2540 generates electron beams that pass through electron beam targeting device 2545 to impinge on slurry 2550 containing cellulosic material. The electron beam targeting device may be a scanner that transmits the beam up to about b feet in a direction approximately parallel to the surface of the slurry 2550 .

On either side of the electron beam emitter 2540 are sonication horns 2530 that deliver ultrasonic wave energy to the slurry 2550 .

The sonication horns 2530 terminate in a removable nozzle 2535 that contacts the slurry 2550 .

The 2530 sonication horns are at risk of damage due to prolonged residual exposure to electron beam exposure. Thus, the horns can be protected by a standard shield 2520, for example, which is made of lead or contains a heavy alloy metal, such as Lipovich metal, which is impermeable to electron beam radiation. However, precautions must be taken to ensure that the presence of the shield does not affect the ultrasonic energy. The removable tips 2535, which are constructed of the same material as the horns 2530 and attached to the horns 2530, normally contact the cellulosic material 2550 and are expected to be damaged. As such, the 2535's removable tips are designed to be easily replaceable.

Another advantage of this simultaneous process of electron beam and ultrasound treatment is that these two processes have complementary results. In the case of irradiation with an electron beam separately, an insufficient dose can lead to cross-linking of some of the polymers in the cellulosic material, which reduces the efficiency of the depolymerization process in general. Also, to achieve a degree of depolymerization similar to the degree of depolymerization achieved using electron beam irradiation and ultrasound treatment separately, lower doses of electron beam irradiation and/or ultrasound irradiation can be used.

The electron beam irradiation device can also be combined with one or more of the high frequency rotor-stator devices that can be used as an alternative to ultrasonic energy devices and can perform a similar function.

Other device combinations are also possible. For example, a device for irradiation with ionizing radiation, which generates gamma radiation emitted, for example, from obo tablets, can be combined with an electron beam source and/or an ultrasonic wave source.

The radiation irradiation devices for biomass pretreatment discussed above can also be combined with one or more devices that perform one or more pyrolysis processing sequences. Such a combination can also have the advantage of high performance. However, caution is necessary, as there may be conflicting requirements between some radiation exposure methods and pyrolysis. For example, sonication devices may require the raw material to be immersed in a liquid oxidizing medium. On the other hand, as discussed above, it may be preferable for a feedstock sample to be pyrolyzed to have a specific moisture content. In this case, new systems automatically measure and monitor specific moisture content and regulate it.

In addition, some or all of the above devices, especially the pyrolysis device, can be combined with the oxidation device as discussed above.

BASIC METHODS

Fermentation

As a rule, various microorganisms can produce a number of useful products by influencing the processed biomass material, for example, its fermentation. For example, alcohols, organic acids, hydrocarbons, hydrogen, proteins, or mixtures of any of these materials can be produced by fermentation or other processes.

A microorganism can be a natural microorganism or an engineered microorganism. For example, a microorganism can be a bacterium, such as a cellulolytic bacterium, a fungus, such as yeast, a plant, or a single-celled organism such as algae, a protozoa, or a fungus-like single-celled organism, such as a slime mold. When organisms are compatible, mixtures of organisms can be used.

To promote the destruction of processed biomass materials, which include cellulose, it is possible

To use one or more enzymes, for example cellulolytic enzyme. In some embodiments, materials that include cellulose are first treated with an enzyme, for example, by combining the materials and the enzyme in an aqueous solution. This material can then be combined with a microorganism. In other embodiments, the combination of materials, which include cellulose, one or more enzymes and a microorganism, is carried out simultaneously, for example by combining in an aqueous solution.

Also, to promote degradation of the treated biomass materials, the treated biomass materials can be further treated (eg, after irradiation) with heat, a chemical reagent (eg, a mineral acid, a base, or a strong oxidizing agent such as sodium hypochlorite), and/or an enzyme.

In the fermentation process, the sugars released at the stage of cellulolytic hydrolysis or saccharification are fermented, for example, into ethanol by a fermenting microorganism such as yeast. Suitable fermenting microorganisms are capable of converting carbohydrates such as glucose, xylose, arabinose, mannose, galactose, oligosaccharides or polysaccharides into fermentation products. Fermenting microorganisms include strains of the genus Zasspgotusez 5yr. example

Zasspgotusev5 segemiziae (baker's yeast), zZasspagotusevz aiaysiv, Zasspagotusev imagit; genus Kinumegotuses, for example species Kipsumegotuses5 taghiapi5, Kipumegotuses iadiiii5; genus Sapaida, for example Sapaida rzeiydoihorisaii5 and Sapaida bBgazzisae; genus Siamizroga, for example species

SiIamizroga Iuzyapiaeye and SiIamizroga oripiae; genus Raspuzoiep, for example species of Raspuzoiep

Iapporpyi5; genus Vgehappotuse5, for example, species Vgeiappotusez siaizepia; the genus Rispia, for example species of Rispia 5iirii5; and the genus Sasspagorpady5, for example, species of Sasspahorpady dedgadapz (RPypyrrids,

S,.R., 1996, SeyiPshiose riosopmegsiop Thiespoiodu, Naparook op Vioeiinapo!: Rhodisiyup apa Shknigayop,

Mutap, S.E., ed., Tauyug 5 ERgapsi5, MMazpipdaip, OS, 179-212).

Commercially available yeasts include, for example, Kea (available from Kea, USA) EBAGIF (available from Riezsptapp' Weaby, Division of Vigp5

Riir Rosy Ips., USA), BOREKZTAKTO) (available from Ashesi, currently I aPietapa), SECT

ZTKAMOF (available from Segi 5igapa AV, Sweden) and REKMOI F (available from OM 5resiaKiev).

Bacteria that can ferment biomass into ethanol and other products include, for example, 2utotopavz tpobBiii5 and Siovigidicht (pegtoseiiYisht (Pirridib5, 1996, above). And e5spipe ei ai. (Iptetaiiopa! Uuoshtpa! oi Zuvivetaiiiys apa Emoishiopagu Misgobiodu 2002, 52, 1155-1160) described anaerobic, mesophilic, cellulolytic bacteria from the forest soil, Siovigidicht rpuiepepiapz 5r. pom., which convert cellulose into ethanol.

Fermentation of biomass to ethanol and other products can be carried out using certain types of thermophilic or genetically engineered microorganisms, such as species

Tpeptpoapaegobasieeg, including T. taiyigapii, and yeast species such as Rispia species. An example of a strain of T. taiyigapi is AZM4, described in bBoppe-Napbzep et al. (Arriiei Mistobiyodu apa

VioyespppoYodu 1993, 38, 537-541) or Apngipu ei a. (Agsp. Misgobrio!. 1997, 168, 114-119).

Yeast and 7utotopap bacteria can be used for fermentation or conversion.

The optimal pH value for yeast is approximately pH 4 to pH 5, while the optimal pH value for 7utotope5 is approximately pH 5 to pH 6. Typical fermentation times are approximately 24 to 96 hours at temperatures ranging from 26 to 40 C, however, thermophilic microorganisms prefer higher temperatures.

Enzymes that break down biomass such as cellulose into lower molecular weight carbohydrate-containing materials such as glucose are called cellulolytic enzymes or cellulases; this process is called "saccharification". These enzymes can be a complex of enzymes that act synergistically, degrading crystalline cellulose. Examples of cellulolytic enzymes include: endoglucanases, cellobiohydrolases, and cellobiases (β-glucosidases). For example, a cellulosic substrate is first hydrolyzed by endoglucanases in random regions, producing oligomeric intermediates. These intermediates are then substrates for exocleaving glucanases such as cellobiohydrolase, with the formation of cellobiose cleaved from the ends of the cellulose polymer. Cellobiose is a water-soluble dimer of glucose bound by a B-1,4 bond. Finally, cellobiase breaks down cellobiose to form glucose.

Cellulase is able to degrade biomass and can be of fungal or bacterial origin. Suitable enzymes include cellulases from the genera Vasii5, Rzendotopav, Nitisoia,

Eizagishtus, TppieYamia, Asgetopishtus, Spguzozorogishtus, and Thispodegipa, and include species of Nyptisoia,

Soriipib5, Tniviamia, Ryvagsht, Museiorpiyoga, Astetopisht, SerNaiIozrogisht, Zsuiaiiidiyt,

Repisiysht or AzregudyShie5 (see, for example, EP 458162), especially cellulases produced by strains selected from the species of Nyitisoia ipboiep5 (reclassified as 5Zsuiaiiidiit

ISheptorpishyt, see, for example, US patent Mo 4435307), Sorgipi5 sipegei5, Ryzagisht ohuzrogyt,

Museiiorpiyoga Iegthornpiia, Megirishv5 adidapiei5, TGNieiamia (etevzigiv, Astetopisht 5r., Astetopisht regvisipit, Astetopisht astetopisht, Asgtetopisht Brgaspurepiit, Astetopisht aispgotozrogyt,

Asgetopisht obsiamayt, Asgetopisht ripKepyopiae, Asgetopisht go5eodgizeit, Asgetopisht ipsoiogaisht iy Asgetopistt yiigaishit; mainly from the species Nitisoia ipboyep5 OM 1800, Rizagiit

З5 охуврогит ОМ 2672, Museiiorpiyoga Sheptorpia SV5 117.65, Serpaiozroitst vr. NUM-202,

Astetopisht 5 years SV5 478.94, Astetopisht 5 years. SV5 265.95, Astetopisht regvisipit SV5 169.65,

Asgetopiyt asgetopisht ANA 9519, Sernaiozrogts v5r. SV5 535.71, Astetopisht Bgaspurepiyt

Sv 866.73, Astetopisht aispgotozrogyt SV5 683.73, Astetopisht orsiamaisht SV5 311.74,

Astetopisht ripkepopide SVB 157.70, Astetopisht gozeodgisent SV5 134.56, Astetopisht ipsoiogaiit SV5 146.62 and Asgetopisht iigaisht SV5 299.70N. Cellulolytic enzymes can also be obtained from Sphagnum sp.

Tpispodegta Copypdia), alkalophilic Vasiyl5 (see, e.g., US Pat. No. 3,844,890 and EP 458,162) and Zigeriotuses (see, e.g., EP 458,162).

It is also possible to use cellulolytic enzymes produced using recombinant technology (see, for example, UMO 2007/071818 and UMO 2006/110891).

The cellulolytic enzymes used can be obtained by fermentation of the indicated strains of microorganisms on a nutrient medium containing suitable sources of carbon and nitrogen and inorganic salts, using methods known in the field (see, for example,

Veppei apa Gavbige, I. (sd5.), Moge Sepe Mapirshiayope ip ERipdi, Asadetis Rgez5, SA 1991).

Suitable media are available from commercial suppliers and can be obtained according to published formulations (eg, in Ategisap Toure Siyige SoiIesiiop catalogs). Temperature ranges and other conditions suitable for the growth and production of cellulase are known in the art (see, e.g., Waiyu apa OiYii5, Viospetisa! Epdipeehipd EipdatepiaI5, MeoOgam/-NIIII VookK

Sotrapu, Moscow State University, 1986).

Processing of cellulose with cellulase is usually carried out at temperatures from 30 to 6576.

Cellulases are active in the pH range from about 3 to 7. The duration of the saccharification stage can be up to 120 hours. The dosage of the cellulase enzyme ensures a fairly high level of cellulose conversion. For example, the appropriate dosage of cellulase, as a rule, is from 5.0 to

50 units of filter paper (ERI or ME) per gram of cellulose. ERI is a standard measure and is defined and measured according to Spoze (1987, Ryge apa Arri. Spect. 59:257-268).

In specific embodiments, the enzyme complex ASSEGEKABEF 1000 is used as an enzyme system in a dosage of 0.25 ml per gram of substrate. Enzyme complex

ASSAY ECABEF 1000 is a cocktail of many enzymes with many types of activity, mainly exogluconase, endogluconase, hemicellulase and beta-glucosidase. The cocktail has a minimum endogluconase activity of 2500 SMS O/g and a minimum beta-glucosidase activity of 400 rMPO O/g. The pH of the cocktail is from about 4.8 to about 5.2. In other specific embodiments, the enzyme system used is a mixture

SEP PHOSIGABZTF 1.5 and Momolute 188. For example, for each gram of substrate, you can use 0.5 ml of SE HOSI ABTF 1.5I and 0.1 ml of Momolute 188. When higher hemicellulase (xylanase) activity is desired, you can use ORTIMAZNOe VO.

Gasification

In addition to the use of pyrolysis for the pretreatment of raw materials, pyrolysis can also be used to process pretreated raw materials to extract useful materials. In particular, a form of pyrolysis known as gasification can be used to produce fuel gases, in addition to various other gaseous, liquid and solid products. For gasification, pre-treated raw materials are fed into a pyrolysis chamber and heated to a high temperature, usually 700 C or more. The temperature used depends on a number of factors, including the nature of the raw material and the desired products.

Also, some amounts of oxygen (eg, as pure oxygen gas and/or as air) and steam (eg, superheated steam) are added to the pyrolysis chamber to promote gasification.

These compounds react with the carbon-containing feedstock material in a multistage reaction to form a mixture of gases called syngas (or "syngas"). Basically, during gasification, a limited amount of oxygen is supplied to the pyrolysis chamber to ensure the combustion of part of the raw material with the formation of carbon monoxide and the generation of process heat. The process heat can then be used to start a second reaction that converts the additional feedstock into hydrogen and carbon monoxide.

In the first stage of the reaction in general, heating the raw material leads to char

Of the residues, which can include a wide variety of different hydrocarbon-based compounds. Certain volatiles (such as certain gaseous hydrocarbon materials) may be formed, resulting in a reduction in the total weight of the raw material. Then, in the second stage of the reaction, part of the volatile material that is formed in the first stage reacts with oxygen in a combustion reaction to form both carbon monoxide and carbon dioxide. In this combustion reaction, heat is released, which triggers the third stage of the reaction. In the third stage, carbon dioxide and steam (for example, water) react with the carbonaceous residues formed in the first stage, with the formation of carbon monoxide and hydrogen gas. Carbon monoxide can also react with steam, in a water vapor conversion reaction, to form carbon dioxide and additional hydrogen gas.

Gasification can be used as the main process to obtain products directly from pre-treated raw materials, for example, for further transport and/or sale. Alternatively or additionally, gasification can be used as an auxiliary process to obtain fuel for the entire processing system.

The hydrogen-rich syngas produced in the gasification process can be burned, for example, to generate electricity and/or process heat, which can be used in other parts of the processing system. As a result, the overall recycling system can be at least partially self-sufficient. A number of other products can also be obtained during and/or after gasification, including pyrolytic oils and hydrocarbon-based gaseous substances; they can be separated and stored or transported if desired.

A number of different pyrolysis chambers are suitable for gasification of pretreated feedstocks, including the pyrolysis chambers described herein. In particular, fluidized bed reactor systems, in which pretreated raw materials are subjected to fluidization in oxygen/air vapor, provide relatively high productivity and direct separation of products. The solid carbonaceous residue that remains after gasification in a fluidized bed system (or other pyrolysis chambers) can be burned to generate additional process heat to drive further gasification reactions.

PROCESSING OF PROCESSED BIOMASS

Distillation

After fermentation, the obtained fluids can be subjected to distillation using, for example, a "fermentation column" to separate ethanol and other alcohols from most of the water and residual solids. The steam leaving the fermentation column can be 35 wt.9o ethanol, and it is fed to the rectification column. A mixture of practically azeotropic (92.5 95) ethanol and water from the distillation column can be purified to pure (99.5 95) ethanol using vapor phase molecular sieves. The sludge from the fermentation columns can be sent to the first stage of a three-stage evaporator. The reflux condenser can provide the heat for this first stage. After the first stage, solids can be separated using a centrifuge and dried in a rotary dryer. Part (255905) of the product from the centrifuge can be reused for fermentation, and the remainder can be sent to the second and third stages of the evaporator. Much of the evaporator condensate can be returned to the process as fairly clean condensate, where a small portion is separated for waste water treatment to prevent the formation of low-boiling compounds.

Treatment of waste water

Wastewater treatment is used to minimize the need for additional water at the plant by recycling the water for reuse at the plant. Wastewater treatment can also provide fuels (such as sludge and biogas) that can be used to increase the overall efficiency of the ethanol production process.

For example, as described in more detail below, sludge and biogas can be used to generate steam and electricity that can be used in various manufacturing processes.

The waste water is first pumped through a screen (such as a grate) to remove large particles that are collected in a waste container. In some embodiments, large particles are sent to landfills. Additionally or alternatively, the large particles are burned to generate steam and/or electricity, as detailed below. Typically, the grid spacing is between 1/4 inch (0.6 cm) and 1 inch (2.5 cm) (eg 1/2 inch (1.3 cm)).

The waste water then flows into an equalization tank, where the concentration of organic compounds in the waste water is balanced during the settling time. As a rule, the settling time is from 8 hours to 36 hours (for example, 24 hours). A mixer for mixing the contents of the container is located in the container. In some embodiments, the implementation is for mixing

From the contents of the container, a set of mixers located throughout the container are used. In certain embodiments, the mixer essentially mixes the contents of the equilibration container so that conditions (eg, wastewater concentration and temperature) throughout the container are uniform.

The first pump pumps water from the equalization tank through the liquid-liquid heat exchanger. The heat exchanger is regulated (for example, by controlling the flow rate of the fluid through the heat exchanger) so that the waste water leaving the heat exchanger has the desired temperature for anaerobic treatment. For example, the desired temperature for anaerobic treatment can be from 40 to 60 "С.

After leaving the heat exchanger, the waste water enters one or more anaerobic reactors. In some embodiments, the concentration of sludge in each anaerobic reactor is the same as the total concentration of sludge in the waste water. In other embodiments, the anaerobic reactor has a higher sludge concentration than the total sludge concentration in the waste water.

A nutrient solution containing nitrogen and phosphorus is measured into each anaerobic reactor containing waste water. The nutrient solution reacts with the bottom in an anaerobic reactor to produce biogas, which can contain 50 95 methane and has a calorific value of approximately 12,000 British thermal units, or Vis, per pound (28,000 kJ/kg). Biogas leaves each anaerobic reactor through a vent and enters the collector, where multiple biogas streams are combined into a single stream. The compressor pumps the biogas stream into a steam boiler or internal combustion engine, as described in more detail below. In some embodiments, the compressor also pumps a single stream of biogas through the desulfurization catalyst. Additionally or alternatively, the compressor may pump a single stream of biogas through a sedimentation trap.

The second pump pumps the anaerobic output stream from the anaerobic reactors to one or more aerobic reactors (for example, rectors for activated sludge). Each aerobic reactor is equipped with an aerator for mixing the anaerobic effluent, the bottom and oxygen (for example, oxygen contained in the air). In each aerobic reactor, the oxidation of cellular material in the anaerobic effluent leads to the production of carbon dioxide, water, and ammonia.

The aerobic outlet stream is moved (for example, by gravity) to a separator, where the sludge is separated from the treated water. A portion of the bottoms is returned to one or more aerobic reactors to create an increased concentration of bottoms in the aerobic reactors, thereby promoting the aerobic destruction of cellular material in the waste water. The conveyor removes excess sediment from the separator. As described in more detail below, excess bottoms are used as fuel to generate steam and/or electricity.

Treated water is pumped from the separator into the sump. Solids dispersed in the treated water settle to the bottom of the sump and are subsequently removed. After the settling period, the treated water is pumped out of the settling tank through a fine filter to remove any additional solids remaining in the water. In some embodiments, chlorine is added to the treated water to kill pathogenic bacteria. In some embodiments, one or more methods of physicochemical separation are used for further purification of treated water. For example, treated water can be pumped through a coal absorption reactor. As another example, treated water can be pumped through a reverse osmosis reactor.

Incineration of waste

The production of alcohol from biomass can lead to the formation of various by-product streams suitable for generating steam and electricity for use in other parts of the plant.

For example, steam generated by burning by-product streams can be used in the distillation process. As another example, electricity generated by burning by-product streams can be used to power electron beam generators and ultrasonic transducers used in pretreatment.

Byproducts used to generate steam and electricity are generated from multiple sources throughout the process. For example, anaerobic digestion of waste water produces biogas with a high methane content and a small amount of waste biomass (scum). As another example, post-distillation solids (eg, unconverted lignin, cellulose, and hemicellulose remaining after pretreatment and core processes) can be used as fuel.

Biogas is fed to an internal combustion engine connected to an electric one

With a generator, for generating electricity. For example, biogas can be used as a fuel source for an electric-ignition natural gas engine. As another example, biogas can be used as a fuel source for a direct injection natural gas engine. As another example, biogas can be used as a fuel source for an internal combustion turbine. Additionally or alternatively, the internal combustion engine can be adapted for a combined configuration to produce electrical and thermal energy. For example, waste heat from internal combustion engines can be used to provide hot water or steam throughout production.

The residue and solids after distillation can be burned to heat the water flowing through the heat exchanger. In some embodiments, the water flowing through the heat exchanger evaporates and is superheated to form steam. In certain embodiments, the steam is used in the reactor for pretreatment and during heat exchange in the processes of distillation and evaporation. Additionally or alternatively, the steam is expanded, powering a multi-stage steam turbine coupled to an electric generator. The steam leaving the steam turbine is condensed with cooling water and returned to the heat exchanger to be reheated to steam. In some embodiments, the rate of flow of water through the heat exchanger is controlled to provide a predetermined production of electricity from a steam turbine coupled to an electric generator. For example, water can be added to the heat exchanger to allow the steam turbine to operate above threshold conditions (for example, the turbine spins fast enough to spin the electric generator).

While some embodiments have been described, other embodiments are possible.

As an example, while the biogas is described as being fed to an internal combustion engine coupled to an electric generator, in certain embodiments the biogas, or a portion thereof, may be passed through a fuel reformer to produce hydrogen. The hydrogen is then converted into electricity using a fuel cell.

As another example, while the biogas is described as being burned separately from the bottoms and solids after distillation, in certain embodiments, some or all of the waste byproducts may be burned together with steam.

PRODUCTS/CO-PRODUCTS

In some embodiments, the present invention relates to materials obtained using the methods described herein. In some cases, such materials can be used in the absence of materials added to the biomass before or after processing, such as materials not naturally present in the biomass. In such cases, the materials will contain naturally occurring materials, such as those derived from biomass.

Alternatively or additionally, the materials obtained using the methods described in this document can be combined with other natural and/or synthetic materials, such as materials that are not naturally present in biomass.

As described above, in some embodiments, the methods described herein can be used to convert (e.g., fermentation) biomass into an energy product (e.g., an alcohol such as ethanol or a hydrocarbon) and/or other products that result from the conversion process. (for example, organic acids). In such cases, biomass is affected by conditions suitable for such conversion. Illustrative conditions may include, for example, at least biomass and one or more microorganisms capable of converting biomass to energy (eg, alcohol) under conditions suitable for the functioning of organisms. The conversion process can be allowed to proceed to the extent that at least some of the biomass is converted to energy (e.g., ethanol) and/or other conversion products (e.g., as described below), and/or to the extent that all (e.g. , essentially all) of the materials converted into energy (eg, ethanol) and/or other products that are formed as a result of the conversion. For example, at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, or 100 95 of the materials subjected to conditions suitable for fermentation are converted to energy (e.g., ethanol ) and/or other products that are formed as a result of conversion.

Alternatively or additionally, the methods described herein can be used to modify the biomass, for example to modify (e.g. increase, decrease or preserve) the solubility of the starting materials, to change the structure, for example to functionalize, the starting materials and/or to change (e.g. , decrease) of molecular weight and/or crystallinity relative to the starting material. Such methods can be carried out together or separately. For example, the methods described in this document can be used to convert part of the biomass into energy. The methods described in this document can also be used to modify (eg, increase, decrease, or save)

From solubility, changes in structure, for example functionalization, and/or changes (for example, reduction) in molecular weight and/or crystallinity of biomass, or vice versa.

In some embodiments, the methods described herein can be used to recover (e.g., extract, isolate, and/or increase the availability, e.g., compared to unprocessed biomass materials) of one or more components contained in the unprocessed biomass material (e.g., source material). Illustrative components that can be recovered (e.g., extracted, isolated, and/or made more available (e.g., compared to unprocessed biomass material)) include, but are not limited to, sugars (e.g., 1,4-dibasic acids (e.g., succinic acid, fumaric acid and malic acid), 2,5-furandicarboxylic acids, 3-hydroxypropionic acid, aspartic acid, glucaric acid, glutamic acid, itaconic acid, 3-hydroxypropionic acid, aspartic acid, levulinic acid, 3-hydroxybutyrolactone, glycerin, sorbitol and/or xylitol/arabit), dextrins, cyclodextrins, amylase, amylopectin, cake, proteins, amino acids, peptides, nucleic acids, lipid fats, fatty acids, gluten, sweeteners (eg glucose), sugar alcohols (eg arabbit, xylitol, ribitol, mannitol, sorbitol, isomaltitol, maltitol and lactitol), oils (e.g. triglyceride vegetable oils (e.g. soybean oil, palm oil, canola oil, sunflower oil, peanut oil cottonseed oil, palm kernel oil, olive oil), corn oil, oat oil, nut oil and palm oil), minerals, vitamins, toxins and other chemicals, ash and flavonoids. Such components can be used in various applications described below, for example as individual components, in combination with one or more additional components, in combination with processed and/or unprocessed biomass, and/or in combination with one or more non-derived additional components ( e.g. extracted, isolated and/or subjected to methods of increasing bioavailability) from biomass. Methods of obtaining one or more of these components are known in the art.

In some embodiments, the methods described herein can be used to increase the availability of one or more components contained in biomass (eg, unprocessed and/or partially processed biomass). Components with increased availability may be more easily obtained (eg, extracted and/or isolated), easier to use, and/or they may be more accessible to the animal (eg, because digestible or absorbed by the animal). Components with increased availability may include, for example, components that occur in biomass under natural conditions and/or components that are formed using methods described herein (eg, cross-linked compounds, low molecular weight compounds). Such components can increase the value of biomass. For example, low molecular weight compounds can be more easily hydrolyzed in the stomach than unprocessed biomass. Thus, biomass containing more accessible low-molecular compounds can be used as a valuable food source, for example for animals or insects, or for applications in agronomy, aquaculture, for example, in the breeding of fish, aquatic microorganisms, aquatic plants, sea grass and algae.

In some embodiments, the methods described herein can be used to sterilize biomass to ensure the suitability of the materials for animal and/or human consumption (e.g., ingestion or implantation), insects, or for applications in agronomy, aquaculture, e.g. in breeding fish, aquatic microorganisms, aquatic plants, sea grass and algae. In some embodiments, treatment of the cellulosic material with irradiation provides sterility of the biomass and, thus, suitability for animal and/or human consumption (eg, for ingestion or implantation). Also, irradiated cellulose can be used in other products or co-products.

In some embodiments, the methods described herein can be used to process biomass into material intended for consumption (eg, ingestion or implantation) by humans and/or non-human animals. As a general rule, such materials should not substantially contain infectious material (e.g. pathogenic and/or non-pathogenic material), toxins and/or other materials (e.g. bacterial and fungal spores, insects and larvae) that may be harmful to humans and/or or animals. Methods known in the art and/or described herein can be used to remove, inactivate, and/or neutralize infectious material (eg, pathogenic and/or non-pathogenic material) and/or toxins that may be harmful to humans and/or animals, or which are generally undesirable in material intended for human and/or animal use. For example, the methods can be used to remove, inactivate and/or neutralize infectious material that may be present in the biomass. Such materials include, for example, pathogenic ones

Zo and non-pathogenic bacteria, viruses, fungi, parasites and prions (for example, infectious proteins). In some cases, the methods described herein can be used to remove, inactivate and/or neutralize toxins, such as bacterial toxins and plant toxins.

Alternatively or additionally, the methods described herein can be used to remove, inactivate and/or neutralize materials that may be present in the biomass, which are not necessarily harmful, but which are undesirable in materials to be used in humans and/or animals or in agronomy or aquaculture. Illustrative materials include, but are not limited to, bacterial and fungal spores, insects and larvae.

In some embodiments, the methods described herein can be used to produce the products and co-products and bioconversion products described herein under adverse conditions. Such conditions may include conditions that represent spatial limitations and/or extreme environmental conditions, such as areas of excessive heat or cold, areas of excessive radiation, areas of excessive pollutants, and/or areas of limited oxygen or sunlight. In some embodiments, such conditions may include, but are not limited to, for example, aboard spacecraft, aboard space stations (e.g., in space), aboard submarines (e.g., nuclear submarines), and other marine vessels or barges or platforms, intended to remain in the sea for extended periods of time, underwater facilities (e.g. civilian and/or military underwater facilities), desert conditions, polar conditions, subzero temperature conditions (e.g. in permafrost areas), conditions high altitudes (eg where oxygen supply may be limited and/or extreme temperatures exist) and remote locations (eg autonomous bases).

In some embodiments, the products and/or co-products described herein, e.g., obtained by processing biomass using the methods described herein, may be, e.g., solids (e.g., particles (e.g., films), granules, and /or powders), semi-solid substances, liquids, gases, vapors, gels and their combinations.

Alcohols

Alcohols produced using the materials described herein may include, but are not limited to, a monohydroxyalcohol, such as ethanol, or a polyhydroxyalcohol, such as ethylene glycol or glycerin. Examples of alcohols that can be produced include, but are not limited to, methanol, ethanol, propanol, isopropanol,

butanol, for example, n-, sec- or tert-butanol, ethylene glycol, propylene glycol, 1,4-butanediol, glycerol or mixtures of these alcohols.

In some embodiments, alcohols produced using the processing methods described herein can be used to produce potable beverages.

Hydrocarbons

Hydrocarbons include aromatic hydrocarbons or arenes, alkanes, alkenes and alkynes.

Illustrative hydrocarbons include methane, ethane, propane, butane, isobutene, hexane, heptane, isobutane, octane, isooctane, nonane, decane, benzene, and toluene.

Organic acids

Organic acids produced using the methods and materials described herein may include monocarboxylic acids or polycarboxylic acids. Examples of organic acids include formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, palmitic acid, stearic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, oleic acid, linolenic acid, glycolic acid, lactic acid acid, γ-hydroxybutyric acid or mixtures of these acids.

Food

As described herein, the present invention relates to methods suitable for modifying biomass, for example, by modifying (e.g., increasing, decreasing, or maintaining) the solubility of the starting materials, changing the structure (e.g., functionalization) of the starting materials, and/or changing (e.g., decrease) of molecular weight and/or crystallinity relative to the starting material. The methods can be used to obtain materials with properties that may be favorable for use as food products or in the production of food products. For example, the methods can be used to produce a material with improved (eg, increased or decreased) solubility, for example, compared to the starting material, which can be used as a more absorbable product. Increased solubility can be provided, for example, by dispersing (eg, dissolving) the unprocessed and processed materials in

With a suitable solvent, removal of undissolved material, detection of materials and/or certain components of materials (eg, sugars) and comparison of levels of detected materials in processed and unprocessed materials. In some cases, the solvent containing the materials can be modified, for example, by heating or pH correction.

Alternatively or additionally, the methods can be used to produce a material with higher nutritional value (eg, higher energy (eg, more digestible dietary energy) and/or nutrient availability) when the material is consumed by the animal, eg, compared to the original material or unprocessed biomass. Such methods will not necessarily increase the total amount of energy or nutrients present in a given amount (eg, mass) of a particular type of processed material compared to the same amount and type of unprocessed biomass.

Instead, the methods described herein can be used to increase the nutritional value (e.g., energy availability and/or one or more nutrients) of a set amount (e.g., mass) of a specific type of processed biomass compared to the same amount and type unprocessed biomass.

Increasing the caloric availability of food in a specific type of biomass can be used to increase the use of metabolizable energy (MEI) of that biomass. Methods of measuring the calorie content of food are known in this field. The MEI is typically calculated by multiplying the number of kilocalories or kilojoules contained in a food component by 8595. In some embodiments, the methods described herein can be used to increase the MEI of biomass.

Ways to compare ME! processed and unprocessed biomass may include, for example, feeding equal amounts of processed or unprocessed biomass to at least two separate groups of one or more animals and measuring the growth response of the animals.

The availability of nutrients can be assessed by carrying out a balance study.

Protocols for carrying out balance studies are known in this field. For example, total nutrient levels can be determined in processed or unprocessed biomass. Equal amounts of processed or unprocessed biomass can be fed to at least two different groups of one or more animals. Then the losses with excrement of one or more nutrients during a given period of time are determined. Increased availability of nutrients is defined as lower amounts of one or more nutrients in animal excrement. Alternatively or additionally, nutrient availability can be assessed by measuring and comparing the levels of one or more nutrients in the blood of animals fed processed and unprocessed biomass.

In some embodiments, nutritional value can be increased by increasing the digestibility of one or more of: dietary energy, carbohydrates, sugars, proteins, fats (saturated, monounsaturated, and polyunsaturated), cholesterol, dietary fiber, vitamins (e.g., vitamin A, E, C, Bb, B12, carotene, thiamine, riboflavin and niacin), minerals (for example, calcium, phosphorus, magnesium, iron, zinc, copper, potassium, selenium and sodium) and oils when the animal consumes biomass.

Generally, the methods described herein can be selected and/or optimized to select a method or combination of methods that results in a more soluble, digestible, and/or absorbable material, e.g., with desirable nutrient availability (e.g., higher nutrient availability substances (eg, proteins, amino acids, carbohydrates, minerals, vitamins, fats, lipids and oils) than in the original unprocessed material) that can be used in humans and/or animals as a food product. Since biomass materials are readily available and inexpensive, materials produced in these ways will provide economical food products and reduce waste.

In some embodiments, the materials and methods described herein can be used to produce foods, such as agricultural foods and foods suitable for consumption by mammals, birds, and/or fish. Such animals include, but are not limited to, food producing animals, domestic animals, zoo animals, laboratory animals, and/or humans.

In some embodiments, the materials produced by the methods described herein that are intended for use as food (eg, human and/or animal) may be further processed, eg, hydrolyzed. Hydrolysis methods are known in the art and include, for example, the use of enzymes, acids and/or bases to reduce the molecular weight of saccharides. In some embodiments, foods produced by the methods described herein may include enzymes (e.g.,

(dry enzymes, active enzymes and/or enzymes requiring activation).

In some embodiments, materials obtained by the methods described herein that are intended for use as food (eg, human and/or animal) may be further processed to enhance the sterility of the materials and/or remove, inactivate, and/or neutralize the materials , which may be present in the biomass, such as infectious material (eg pathogenic and/or non-pathogenic material), toxins and/or other materials (eg bacterial and fungal spores, insects and larvae). Generally, the methods described herein can be selected and optimized to provide optimal removal, inactivation, and/or neutralization of materials that may be undesirable.

Animal feed

More than 600 million tons of animal feed are produced worldwide each year, with an annual production growth rate of approximately 2 95. Agriculture is one of the largest consumers of animal feed, and US farmers spend more than $20 billion annually on agricultural feed for animals intended for for food products. Other feed consumers include, for example, pet owners, zoos, and laboratories that keep animals for scientific research.

As a general rule, animal feed must meet or exceed the specific requirements of a given animal, for example to maintain or improve the health of a particular type or species of animal, to ensure the growth of a given animal (e.g. increase in tissue mass) and/or to stimulate production products. Improved animal feeds (eg, more soluble, absorbable and/or digestible feeds) provide or stimulate these effects using less feed and/or at lower costs.

Currently used feedstocks in commercially produced feeds include feed grains (eg, corn, soybeans, sorghum, oats, and barley). The fodder industry is the largest buyer of corn, fodder grain and soybean meal in

USA. However, with the rising cost of feed grains such as corn, cheaper alternatives are desirable. The most widely available feed is biomass, such as cellulosic material. In some embodiments, the methods described herein can be used to increase the availability of nutrients in one of these 60 materials, e.g., to maintain or improve the health of a particular type or species of animal, stimulate the growth of that animal (e.g., increase tissue mass ) and/or to stimulate food production. The low nutrient availability of commonly used forages (for example, hay and grasses) is often associated with the high content of cellulose, hemicellulose and lignin in such material. Unlike humans, who cannot digest cellulose, herbivores, such as ruminants, are able to digest cellulose, at least in part, through a process known as rumination. However, this process is insufficient and requires many rounds of regurgitation. For example, ruminants digest only 30-50 90 cellulose and hemicellulose. In some embodiments, the methods described herein can be used to increase the availability of nutrients or the nutritional value of any of these materials, e.g., to maintain or improve the health of a particular type or species of animal, to stimulate the growth of that animal (e.g., tissue mass increase) and/or to stimulate the production of food products. In the methods described in this document, reduced amounts of feed are used, at lower costs and/or with less waste.

As a rule, increasing the availability of nutrients in animal feed provides a reduction in the amount of feed required to feed an animal so that the animal receives the same amount of energy. Consequently, less feed is required for the animal, thus providing a more economical feed.

Various methods have been used to increase the availability of feed nutrients with limited success. Such methods include the use of enzymes, such as cellulosic enzymes, to break down cellulosic material into shorter-chain oligosaccharides that are more easily digested. Although this practice is used in Europe and Australia, it is expensive and is not often used in developing countries. Other methods include removal of thatch to prevent leaf loss, deaeration, physical treatment of the material (eg compaction of cellulosic material, reduction of particle size and fine grinding), chemical treatment and refeeding. In addition, feeds consisting primarily of cellulosic material are often supplemented with nutrient systems (eg, premix additives). These feeding systems, as a rule, are designed to meet the food needs of a given animal. Despite the guarantee that

Necessary nutrients are obtained from the animal, cellulose material is used inefficiently in such systems.

The methods described herein provide methods for increasing the availability of nutrients or the nutritional value of biomass (e.g., by modifying (e.g., increasing, decreasing, or maintaining) the solubility of the biomass and/or changing the structure (e.g., functionalization) of the starting materials, and/or changing (for example, a decrease in) molecular weight or crystallinity) as described above, thereby obtaining more valuable feeds. In some embodiments, the methods described herein can be used to increase the availability of biomass nutrients by breaking down cellulosic material (eg, cellulose and/or hemicellulose) into shorter chain saccharides or monosaccharides. By increasing the nutrient availability of the biomass, these methods result in a more efficient feed that can be used to maintain or improve the health of a particular type or species of animal, to stimulate the growth of that animal (eg, increased tissue mass) and/or to stimulate food production .

In some embodiments, the beneficial animal feed may include partially processed biomass, such as biomass that has been shredded using the methods described herein. Such partially processed biomass can be more easily hydrolyzed in the animal's stomach.

In some embodiments, the methods described herein can be used to process biomass to produce the materials described herein. These materials may include, but are not limited to, for example, polysaccharides greater than 1,000 saccharide units in length; approximately 1000 saccharide elements; approximately 800-900 saccharide elements; approximately 700-800 saccharide elements; approximately 600-700 saccharide elements; approximately 500-600 saccharide elements; approximately 400-500 saccharide elements; approximately 300-400 saccharide elements; approximately 200-300 saccharide elements; approximately 100-200 saccharide elements; 100, 90, 80, 70, 60, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11,10,9, 8, 7, 6, 5, 4, 3, 2 and 1 saccharide element.

In some embodiments, the methods lead to the formation of disaccharides (for example, sucrose, lactose, maltose, trehalose and cellobiose). In some embodiments, the methods lead to the formation of monosaccharides (for example, glucose (dextrose), fructose, galactose, xylose and ribose). These shorter-chain molecules are more easily absorbed by animals, which,

thereby increasing the availability of biomass nutrients. Therefore, the methods and materials described in this document can be used as feed or in the production of feed.

In some embodiments, the materials described herein can be used as feed, such as agricultural feed and/or feed suitable for consumption by mammals, birds and/or fish. Alternatively or additionally, the methods described herein can be used to process the raw material suitable for use as animal feed or in animal feed.

Materials that can be effectively processed using the methods described herein include cellulosic and lignocellulosic materials, such as arable land products, crops, grasses, plants and/or feed grain, for example, including but not limited to plant material (e.g. forages such as alfalfa meal, hay, Bermuda grass hay, fescue, corn plant and soybean hay), grains (e.g. barley, maize (including natural and genetically modified maize), oats, rice, sorghum and wheat), plant protein products (e.g., canola meal, cottonseed cake and meal, safflower meal, and soybean (including natural and genetically modified soybean) feed and flour), grain by-products (e.g., dry sorghum products, dried brewer's grain , corn gluten, cake and sorghum germ meal, peanut shell and wheat bran), fruit and fruit by-products (for example, in dried citrus pulp, apple cake and pectin pulp), molasses (for example, beet, citrus, starch and cane molasses), almond hulls, ground shell, buckwheat hulls, legumes and legume by-products, and by-products of other crops. Other feedstocks include, but are not limited to, alfalfa, barley, ryegrass, cabbage (e.g., kohlrabi, kale, rapeseed (canola), rutabaga (Swedish turnip) and turnip), clover (e.g., red and white clover, red clover, subterranean clover, and white clover), grasses (e.g. French ryegrass, fescue, Bermuda grass, camphor, sedge, meadow sedge, common hedgehog, weeds, and timothy meadowsweet), corn (corn), ryegrass, millet, sorghum, and soy In some embodiments, the starting material may be livestock waste (eg, ruminant waste) or human waste.

In some embodiments, the feed contains only materials obtained using the methods described in this document. Alternatively or additionally, the feed contains additional starting materials (including starting materials not processed using the methods described in this document) and additives. The composition of the feed can be such that it meets the needs of a given animal, for example, to maintain or improve the health of a particular type or species of animal, to stimulate the growth of a given animal, to increase tissue mass and/or to stimulate food production. In some cases, feed can be made to meet the needs of a given animal at the lowest cost ("least cost ration").

The methods of determining the composition of feed and ration of the least cost are well known to specialists in this field (see, for example, Rebii apa MiSheg, Apita! Reei ERogtshiiaiop: Esopotis apa Sotrshiieg

Arriisayopz (Riapi apa Apita! Zsiepse), Zrgipdeg Rybiizpipuo, Rerbgoagu 28, 1993 and zsher-address of the world wide network Imeiptogtayisv5.sot).

Additional starting materials and additives that can be effectively combined with material obtained using the methods described herein include, but are not limited to, animal products (eg, animal meat, feed scraps, meat and bone meal, poultry meal, animal by-product meal, dried animal blood, blood meal, leek meal, eggshell meal, whole hydrolysed bird, hydrolysed wool and bone marrow), animal waste, seafood and their by-products (e.g. , krill, fish parts and fish meal, fish waste meal, crab parts and crab meal, shrimp parts and shrimp meal, fish oil, fish liver and gland meal, and other fish by-products), dairy products (eg cow's milk powder, casein, whey products and cheese powder), fats and oils (e.g. animal fat, vegetable fat or butter and hydrolysed fats), restaurant food waste (e.g. food waste from restaurants, bakeries and cafeterias) and contaminated/spoilt products treated to kill pathogens.

Other additives include antibiotics (eg, tetracyclines, macrolides, fluoroquinolones, and streptogramins), flavorings, brewer's yeast, pharmaceutical by-products (eg, spent mycelium and fermentation products), minerals and trace elements (eg, bone char, calcium carbonate, chalk, iron salts, magnesium salts, stomatal shell meal and sulfates), proteinized minerals (eg, 60 proteinized selenium and chromium), vitamins (eg, vitamin A, vitamin 0, vitamin B12, niacin, and betaine), direct food organisms /probiotics (for example, Azreguyy5 pideg, Vassyyye5 v5y,yBiyiv,

Vipaobasiegisht apitai!yykh, V. Biydisht, Epiegosossib5 iaesisht, Avregaiy5 ogugaye, Iasiobasiyiv5 asidoripyi5, GG. BiiYidagisy5, G. riapeiagisht, Zigeriosossi5 Iasiykh and Zasspagotusev5 sehemiziaye), prebiotics (eg, mannan-oligosaccharides (MOB), fructo-oligosaccharides and mixed oligodextran), flavors (eg, aloe gel concentrate, ginger, capsicum and fennel), enzymes ( (e.g. phytase, cellulase, lactase, lipase, pepsin, catalase, xylanase and pectinase), acetic acid, sulfuric acid, aluminum salts, dextrans, glycerin, beeswax, sorbitol, riboflavin, preservatives (e.g. butylated hydroxyanisole and sodium bisulfite), nutraceuticals (e.g. herbal and botanical products), amino acids, reserve protein, urea, molasses, fatty acids (e.g. acetic, propionic and butyric acid) and metabolic modifiers (e.g. somatotropins and adrenergic agonists). In some cases, the materials produced using the methods described in this document can be combined or included in a block of urea, molasses and minerals (UMMV).

Feeds made using the materials described in this document may be in a form suitable for consumption, for example, by a given animal. In some cases, the feed may be solid. Alternatively or additionally, the feed may be in liquid form, for example the feed may be in the form of a liquid suspension or solution in a suitable solvent.

Illustrative forms include, but are not limited to, solid forms such as powders, tablets, mineral blocks, granules, briquettes, and mixtures of unprocessed source material (eg, grass) and material processed using methods described herein.

In some embodiments, the materials described herein may be incorporated (eg, mixed) into feed by a farmer, for example, for local use and/or small-batch distribution. In such cases, the materials described herein may be provided to the farmer in packaged form, such as in a form suitable for incorporation into feed. Alternatively or additionally, the materials described herein may be included (eg, mixed) in feed by a feed manufacturer, for example, for large-scale distribution. In such cases, the materials described herein may be provided to the feed manufacturer in a form suitable for incorporation into the feed. Alternatively or additionally,

The materials described in this document can be obtained from the source material in the region where the feed is manufactured.

In some embodiments, the materials described herein can be distributed and consumed by the animal alone in the absence of any additional starting materials and/or additives.

In some embodiments, further processing of the materials is required prior to use as a food product. For example, a dryer can be used to remove moisture from fermentation intermediates to facilitate storage, processing and increase shelf life. Additionally or alternatively. the materials can be ground to a fine particle size in a stainless steel mill to produce a flour-like substance.

As a rule, biomass-based feed is usually fed only to ruminants that are able to digest cellulose, at least partially. Since this description relates to materials in which cellulosic material is broken down into shorter chain sugars, these materials can also be used as acceptable feed for animals that are unable to digest cellulose or hemicellulose. Thus, feeds obtained using the materials and methods described herein can be effectively fed to animals, including, but not limited to, food producing animals, zoo and laboratory animals, and/or pets. Feed can also be used in agronomy and aquaculture. In addition, since the feed obtained using the materials described in this document has a higher availability of nutrients, less feed is required for the animal to obtain the same amount of energy, which can reduce the cost of the feed.

Alternatively, animals may be able to consume greater amounts of energy, resulting in higher growth rates, increased tissue mass, milk production, and egg production.

In some embodiments, the materials described herein can be effectively fed to ruminants (eg, cattle, goats, sheep, horses, crops, bison, deer, camels, alpacas, llamas, giraffes, yaks, buffaloes, wildebeests, and antelopes ), birds, pigs, wild boars, birds, cats, dogs and fish.

Dry liquor and dissolved substances can be converted into a valuable by-product of the distillation-dehydration process. After the distillation-dehydration process, the dry bard and dissolved substances can be dried to improve the storage or processing of the material. The resulting dry bran and solutes (005) are low in starch, high in fat, high in protein, high in fiber, and high in phosphorus. Thus, for example, OO may be valuable as a feed source for animals (eg as a feed source for dairy cattle). BOS can be further combined with nutritional supplements to meet the specific nutritional needs of specific categories of animals (for example, the balance of digestible lysine and phosphorus for pig diets). Alternatively or additionally, biomass processed using the methods described in this document can be combined with OO.

The ratio of processed biomass to BOS can be optimized to meet the needs of a particular animal.

In some embodiments, animal feed can use oils obtained from biomass using the methods described herein, for example, as an additive to pet food.

In some embodiments, the biomass obtained using the methods described herein can be used in animal feed.

Food products for humans

As described above, humans are generally less able to digest cellulose and cellulosic material. Biomass is a widely available material, however, it could serve as a new product for human consumption. However, in order for a biomass material (eg, cellulose-containing material) to be suitable as a human food, the availability of biomass nutrients must be increased by (1) increasing the solubility of the biomass; (2) changes in the structure (for example, functionalization) of the source materials; (3) changes (for example, a decrease) in molecular weight and/or crystallinity relative to the starting material; and/or (4) breaking down cellulosic material into smaller saccharides, such as saccharides longer than 1000 saccharide units; approximately 1000 saccharide elements; approximately 800-900 saccharide elements; approximately 700-800 saccharide elements; approximately 600-700 saccharide elements; approximately 500-600 saccharide elements; approximately 400-500 saccharide elements; approximately 300-400 saccharide elements; approximately 200-300 saccharide elements; approximately 100-200 saccharide elements; 100, 90, 80, 70, 60, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11,10, 9, 8, 7, 6, 5, 4, 3, 2 and 1 saccharide element. Such materials may have increased availability of nutrients (as described above), for example in humans, and may be suitable as human food. As a general rule, a suitable human food product should, for example, provide an acceptable and affordable source of energy and nutrients for humans, for example, to maintain or improve human health and/or stimulate human growth (eg, increase tissue mass). The methods described in this document can be used to produce such useful food products for humans, for example, from biomass-based material.

In some embodiments, processing of the materials is required prior to use as food products. For example, a dryer can be used to remove moisture from fermentation intermediates to facilitate storage, processing and increase shelf life. Additionally or alternatively, the materials can be ground to a fine particle size in a stainless steel mill to produce a flour-like substance.

Such foods may include, but are not limited to, for example, energy supplements (eg, powders and liquids). Alternatively or additionally, the materials described herein can be combined with a first food product to increase the nutritional value of the first food product. For example, the foods described herein can be combined with a low-energy food to increase the energy in that food.

Alternatively or additionally, the materials described herein can be used to enhance the sweet taste of a food product, for example as a sweetener, as well as the nutritional value of the food product. In such cases, it may be desirable to obtain from the materials one or more specific sugars (eg, monosaccharides, disaccharides, oligosaccharides, and/or polysaccharides), for example, by isolating one or more specific sugars from the materials. Methods of extracting sugars are known in this field.

In some embodiments, the materials described herein can be used as a low-cost material for food products.

For example, the materials can be supplied to bakeries for use in bread and/or confectionery, and to food manufacturers for use as a filler, for example to increase the volume and/or nutritional value of a food product.

In some embodiments, the materials may also serve as a source of fiber for human consumption. In such cases, the methods used to break down the cellulolytic material can be adapted to less completely reduce the molecular weight, for example, the methods can lead to materials that contain little cellulose and/or lead to longer chain polysaccharides that are not easily absorbed by humans. . Such materials may be administered to a human in the form of a solid (eg, a tablet or granular powder) or liquid (eg, a solution, gel, colloid, or suspension) for human consumption.

In some embodiments, the materials described herein can be provided to a human for consumption alone or in combination with a second food product that is suitable for human consumption. Such foods include, but are not limited to, bread, dairy products, meat products, fish products, grain products, fruits, vegetables, legumes (eg, soy), and chewing gum. In some embodiments, the materials described herein can be combined with proteins, fats, carbohydrates, minerals, pharmaceuticals, and vitamins.

Squirrels

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate and/or purify) proteins (eg, polypeptides, peptides and amino acids) from biomass. Such proteins (for example, polypeptides, peptides and amino acids) can be used, for example, alone or in combination with one or more materials and components of biomass obtained using the methods described in this document, in the food industry (for example, as additives, excipients and/or fillers), in the cosmetic industry (e.g. in the manufacture of cosmetics) and/or in agronomy (e.g. as nutrients to feed or maintain agricultural crops) or in aquaculture.

In some embodiments, the methods described herein can be used to produce proteins (e.g., polypeptides, peptides, and amino acids) from, for example, okra, Hiripa tishabii, nuts (e.g., macadamia nut), Uezzepia Baiatsa, Oeposagriv, ZiocKevzia Iaemi, and others. , Megopia daiatepviv and Arodapipega ipashiaie.

Fats, oils and lipids

Fats consist of a wide group of compounds that are usually soluble in organic solvents and mainly insoluble in water. Fats are solid at room temperature. Fats that are liquid at room temperature are usually called oils. The term lipids generally refers to solid and liquid fats. As used herein, the terms fats, oils, and lipids include, but are not limited to, edible oils, industrial oils, and materials that include an ester, such as a triglyceride and/or a hydrocarbon.

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate and/or purify) fats (eg, lipids and fatty acids) from biomass. Such fats (e.g., lipids and fatty acids) can be used, for example, alone or in combination with one or more biomass materials and components obtained using the methods described herein in the food industry (e.g., as additives, excipients and /or fillers), in the cosmetic industry (for example, in the manufacture of cosmetic products) and/or in agronomy (for example, as nutrients).

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate and/or purify) oils from biomass. Such oils can be used, for example, alone or in combination with one or more biomass materials and components obtained using the methods described in this document, in the food industry (for example, as additives, excipients and/or fillers), in the cosmetic industry (e.g. in the manufacture of cosmetics), in agronomy (e.g. as nutrients), as biofuels, dry oils (e.g. in paints) and additives in pet food.

In some embodiments, the methods described herein can be used to obtain fats, oils, and/or lipids, for example, from sunflower, okra seed, buffalo squash (Sysygya toeiidibbvita), Igiripi5 tsabiii5, nuts (e.g., macadamia nuts), Ue55epia raiatsa, Oeeposagri5, Statre abuzvzipisa (catran), Mopoesioib5 |oioba (jojoba), Stysitegaye 5r. (for example, Vgazvzisa |ipsea, V. sagipaga, V. parakh (ordinary rapeseed) and V. satrevigiv), ZiokKevzia Iaemi5, Megopia daiatepviv and Arodapipega ipashiaie.

Carbohydrates and sugars

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate and/or purify) carbohydrates and/or sugars from biomass. Such carbohydrates and sugars can be used, for example, alone or in combination with one or more biomass materials and components obtained using the methods described herein, for example, in the food industry (for example, as additives, excipients, syrups and/or fillers), in the cosmetic industry (e.g. in the manufacture of cosmetics) and/or in agronomy (e.g. as nutrients).

Vitamins

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate and/or purify) vitamins from biomass. Such vitamins can be used, for example, alone or in combination with one or more materials and components of biomass obtained using the methods described in this document, for example, in the food industry (for example, as additives and excipients), in the health care industry. I, in the cosmetic industry (for example, in the manufacture of cosmetics) and/or in agronomy.

Minerals

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate, and/or purify) minerals from biomass. Such minerals can be used, for example, alone or in combination with one or more materials and components of biomass obtained using the methods described in this document, for example, in the food industry (for example, as additives and excipients), in the health industry. I, in the cosmetic industry (for example, in the manufacture of cosmetics) and/or in agronomy.

Ash

In some embodiments, the methods described herein can be used to obtain (eg, extract, isolate and/or purify) ash from biomass. Such ash can be used, for example, alone or in combination with one or more materials and components of biomass obtained using the methods described in this document, for example, in the food industry (for example, as an additive, auxiliary substance and/or filler).

Pharmaceuticals

More than 120 currently available pharmaceutical products are of plant origin.

Since the methods described herein are suitable for the processing of cellulolytic material, these methods may be suitable for the isolation, purification and/or preparation of plant-based pharmaceuticals.

In some embodiments, the materials described herein may have medicinal properties. For example, the methods described in this document can lead to materials with new medical properties (for example, not present in the original material). Alternatively or additionally, the methods described herein may result in a material with enhanced medical properties (for example, an increased medical property compared to the starting material).

In some embodiments, the methods described herein can be used to modify (eg, increase, decrease, or maintain) the solubility of a material, such as a material with medical properties. Such material may be more easily administered and/or may be more easily absorbed, for example in humans and/or animals, than the parent material.

In some embodiments, the methods described herein can be used to functionalize (eg, alter the structure, expose a reactive side chain, and/or modify the charge) a material with medical properties. Such materials may have, for example, altered reactivity, altered charge, and/or altered solubility.

In some embodiments, the methods described herein can be used to modify the molecular structure of a material, such as a material with medical properties. Such materials may have altered (eg, increased or decreased) average molecular weight, average crystallinity, surface area, and/or porosity. Such materials may have, for example, altered reactivity, altered charge, altered solubility.

In some embodiments, the methods described herein can be used as highly efficient processing methods, for example, to produce plant-based pharmaceuticals from cellulosic source material, such as plants. In some embodiments 60, the methods described herein can be used to enhance the pharmaceutical activity of a plant-based pharmaceutical. For example, in some embodiments, the methods described in this document can be applied to plants and or herbs with medicinal properties. For example, sonication can stimulate the bioactivity and/or bioavailability of medical components of plants and/or herbs with medicinal properties. In addition or alternatively, irradiation may stimulate the bioactivity and/or bioavailability of medicinal components of plants and/or herbs with medicinal properties.

In some embodiments, the methods described herein can be used to increase the solubility of plant and/or herb material. Alternatively or additionally, the methods described herein can be used to reduce the toxicity of plant and/or herb material without impairing the medicinal properties of the plant and/or herb. In some embodiments, the methods described herein are suitable for isolating and/or purifying pharmaceutical compounds from plant material (which, without regard to theory, is possible due to more efficient degradation of cellulosic material) because the methods destroy, alter, modify, or restructure cellulose, for example, present in the leaves of plant material. Then, the desired compounds released using the methods described herein can be isolated from the undesired material, while less efficient methods may not allow the release of the desired material from the undesired material. Thus, less effective methods inevitably result in the transfer of unwanted material that may reduce the desired efficacy (eg, of a pharmaceutical compound) and/or may be associated with potentially toxic side effects. Thus, the methods described herein can be used to obtain highly purified forms of potential pharmaceutical compounds, for example, that do not contain unwanted plant material, which are not achievable using current methods. These highly purified compounds may be more effective than less purified forms of the same compounds. In some embodiments, the increased efficiency achievable using the methods described herein may allow for reduced dosage. In turn, this reduction in the amount of material administered to the subject may reduce the associated toxicity. Alternatively or additionally, removal of excess or unwanted plant material may help reduce or eliminate any toxicity,

From associated with a plant compound that was subjected to processing using the methods described in this document.

Examples of effective treatment of plants and/or plant material using the methods described herein include, for example, sonication and irradiation, which may be combined with pretreatment of willow bark to stimulate the release, purification, and/or production of salicin. Alternatively or additionally, the methods described herein can be used to process plant material containing fennel plants to facilitate the isolation, purification and/or production of allantoin. Alternatively or additionally, the methods described herein can be used to facilitate the isolation, purification and/or production of benzoin. Alternatively or additionally, the methods described herein can be used to process plant material containing camphor cornflowers to facilitate the isolation, purification and/or production of camphor. Alternatively or additionally, the methods described herein can be used to process plant material containing Erpeaga plants to facilitate the isolation, purification and/or production of ephedrine. Alternatively or additionally, the methods described herein can be used to process plant material containing Juiboiivzia tuorogoide5

E. Vg. (Australian cork tree), to facilitate the isolation, purification and/or production of atropine. In some embodiments, atropine obtained using the methods described herein may have an enhanced anticholinergic effect. Alternatively or additionally, the methods described in this document can be used to process plant material containing Mysypa aegipdiapa (velvet beans) to facilitate the isolation, purification and/or production of I-dopa. In some embodiments, I-dopa obtained using the methods described herein may have an increased antiparkinsonian effect. Alternatively or additionally, the methods described in this document can be used to process plant material containing RVuzovidta mepepozut Vaii. (Calabar bean) to facilitate the isolation, purification and/or production of physostigmine. In some embodiments, physostigmine obtained using the methods described herein may have an enhanced anticholinesterase effect. Examples of other plant-based pharmaceuticals for which the methods described herein can be used to process plant material to isolate, purify 60 iMabo products include, but are not limited to, bromelain, chymopapain, cocaine,

deserpidine, emetine, hyoscyamine, kavaine, monocrotaline, ouabain, papain, pilocarpine, quinidine, quinine, rescinamine, reserpine, scopolamine, tubocurarine, vinblastine, yohimbine, caprylic acid, cineole, citric acid, codeine, cresol, guaiacol, lecithin, menthol, phenolpseudoephedrine, sorbitol and tartaric acid.

In some embodiments, the methods described herein can be used to process herbs, such as medicinal herbs, including, but not limited to, cornflowers, lemon sorghum, parsley, peppermint, and celery. Additional medicinal herbs that can be processed using the methods described in this document can be found at the web address of the world wide web aipaiige.sot.

The latest technology is the production of pharmaceuticals in plants. Pharmaceuticals produced using plants, commonly referred to as plant-produced pharmaceuticals (PPRs), include pharmaceutical compounds and vaccines. As a rule, PMPs are expressed in the leaves of the corresponding plants. Thus, it is apparent that the methods described herein may be useful for processing plant material containing PMP to facilitate the isolation, purification and/or production of PMP.

Additional illustrative medicinal plants that can be processed using the methods described in this document can be found, for example, at the World Wide Web URL pr5.dom/riapiz/MEOVISIMAI/riapiv5. sweat.

In some embodiments, the material processed using the methods described herein can be combined with a pharmaceutical excipient, for example, for administration to a subject. Illustrative excipients that can be used include buffers (eg, citrate buffer, phosphate buffer, acetate buffer, and bicarbonate buffer), amino acids, urea, alcohols, ascorbic acid, phospholipids, polypeptides (eg, serum albumin), EDTO, sodium chloride, liposomes , mannitol, sorbitol, water and glycerin. Dosage forms can be made to be suitable for any standard route of administration. For example, administration may be parenteral, intravenous, subcutaneous, or oral, or it may be any route of administration approved by Eedega! The abomination of Ayatipivigayop (see Mayor's address on the World Wide Web

Ttda.dom/sdeg/dzt/Yu V s/ag900301.pit).

From Nutraceuticals and nutraceuticals

Products with medical health benefits, including disease prevention and/or treatment, are called nutraceuticals or nutraceuticals. For example, nutraceuticals and nutraceuticals are naturally occurring or artificially derived nutritional supplements that can promote a healthy lifestyle, for example, by alleviating disease-related symptoms, reducing the incidence or severity of disease, and promoting long-term health.

In some embodiments, the methods described herein can be used to produce combinations of monosaccharides, disaccharides, oligosaccharides, and/or polysaccharides that can promote a healthy lifestyle. In some embodiments, the methods described herein can be used to produce material that is suitable for weight loss in humans. For example, a material such as a fibrous material may have low availability of nutrients with low digestibility. Such materials can be used as a supplement to the diet, for example, to suppress hunger or ensure satiety. Consumption of such materials may allow the subject to avoid consuming highly available nutrients and/or highly digestible foods and thus may contribute to weight loss in the individual.

In some embodiments, the materials described herein can be supplemented with one or more nutritional supplements that can support a healthy lifestyle.

In such cases, the materials described herein can either enhance the activity of one or more nutritional supplements and/or increase the solubility and/or enhance the pharmacokinetic properties of the nutritional supplements. Illustrative nutritional supplements that may be combined with the materials described herein include, but are not limited to, e.g., silicon dioxide, silicon, boron, potassium, iodine, beta-carotene, lycopene, insoluble fiber, monounsaturated fatty acids, omega- C-fatty acid, flavonols, sulforaphane, phenols (e.g. caffeic acid and ferulic acid), plant stanols and sterols (including their esters), polyols (e.g. sugar alcohols), prebiotics and probiotics (e.g. lactic acid bacteria and bifidobacteria) , phytoestrogens (eg, isoflavones such as daidzein and genistein), proanthocyanids, soy protein, sulfides and thiols (eg, dithiolthiones), vitamins (eg, vitamin A, vitamin VI, vitamin B2, vitamin B3, vitamin B5, vitamin BbE, vitamin 60 B7, vitamin B12, vitamin C, vitamin 0, vitamin E, vitamin K, including their combinations), minerals

(eg, iron, calcium, magnesium, manganese, phosphorus, potassium, zinc, trace elements, chromium, selenium, including combinations thereof) and folic acid.

Pharmaceutical dosage forms and compositions for the delivery of medicines

Medicinal substances are rarely administered alone, rather as part of a formulation in combination with one or more non-medicinal agents that perform different and often specialized pharmaceutical functions. Pharmaceutics is the science of the development of dosage forms, for example, the manufacture of medicinal products in the form of a dosage form suitable for administration to a subject. These non-medicinal agents, called pharmaceutical substances or pharmaceutical ingredients, can be incorporated into the composition to solubilize, suspend, thicken, dilute, emulsify, stabilize, preserve, color, flavor, and shape medical products to produce effective and attractive dosage forms.

Such dosage forms can be unique in their physical and pharmaceutical characteristics. The drug and pharmaceutical ingredients are generally compatible with each other to produce a drug product that is stable, effective, attractive, safe, and easy to administer. The product must be manufactured under appropriate quality control measures and must be packed in containers that provide enhanced stability. Methods describing the preparation of specific dosage forms are well known in the art and can be found, for example, in Apzei et al., Rnappaseciisa! bozade ERogtv apa

Ogyd Oeiiimegu Buzietv, zemepiy Eainop, I irripsoii, UMiPatv, in "UMiKipe Vetipdiopye Rpagtaseciisai

Zsiepsev", 181p Ed., Seppago, ed., Mask Rybiizpipu So., Eazyup, Ra., 1990.

In some embodiments, the pretreated materials described herein can be used as pharmaceutical ingredients, such as inactive ingredients. For example, the materials described herein can be used for the manufacture, for example, of solubilizing, suspending, thickening, diluting, emulsifying, stabilizing, preserving, coloring, flavoring, and shaping medicinal substances into effective and attractive dosage forms that have a pleasant taste. . In such cases, the materials described herein can be mixed with the drug and/or conjugated with the drug so that the solubility, concentration, viscosity, emulsion stability, shelf life, color and taste of the drug are increased or decreased.

In some embodiments, the methods described herein can be used to modify (eg, increase, decrease, or maintain) the solubility of a material.

Such materials can be used to facilitate administration of a drug to a subject.

For example, some of the new pre-processed materials are exclusively soluble in liquids such as water and can be used by mixing with the active ingredients to produce a pharmaceutical composition so that the inert ingredients are readily dissolved in the liquids.

Alternatively or additionally, the materials described herein can be used to slow, control, or modify the release of a drug after administration of the drug to a subject. In such cases, the materials described herein can be used in solid dosage forms and/or controlled release drug delivery systems; semi-solid and/or transdermal systems; pharmaceutical inserts; liquid dosage forms; sterile dosage forms and delivery systems; and new and improved dosage forms, delivery systems and devices. For example, the materials described herein can be formulated, for example, in the form of a tablet, capsule (eg, hard capsule, soft capsule, or microcapsule), suppository, injectable (parenteral) solution or suspension, cream, ointment, ophthalmic solution or suspension, eye drop solution or suspension, inhaled solution or suspension, nasal spray, transdermal patch, emulsion, ointment, cream, gel, suspension, dispersion, solution (eg, intravenous solution), implant, implant coating, lotion, pill , gel, powder and paste. In some embodiments, the materials described herein can be combined with a radiopharmaceutical.

In some embodiments, the methods described herein can be used to produce a material that can be conjugated to a biological agent and/or a pharmaceutical agent. Such conjugates can be used to facilitate the introduction of the agent and enhance the pharmaceutical properties of the agent.

Formulations and methods of administration can be adapted for the disease or disorder being treated and for the particular individual being treated. When using the materials described in this document as pharmaceutical ingredients, it may be necessary to determine the optimal composition and type of dosage. For example, 60 different starting formulations can be developed and tested for desired characteristics (eg, drug release profile, bioavailability and clinical efficacy) and for preliminary research into their production and product scale-up. The formulation that best meets the product objectives (eg, drug release profile, bioavailability, and clinical efficacy) can then be selected as the master formulation. Each subsequent batch of product can then be manufactured to meet the requirements of the master formula. For example, if the product is intended for systemic use and oral administration is desired, tablets and/or capsules are usually made. Also, when choosing a dosage form, you can take into account the age of the intended patient. For example, for infants and children under five years of age, pharmaceutical liquids are preferred for oral administration, rather than solids. In addition, the physical characteristics of the drug or drugs to be included in the formulation together with the pharmaceutical ingredients must be understood prior to the development of the dosage form.

Pharmaceutical compositions containing one or more compounds described herein can be prepared according to the intended route of administration.

In some cases, the nature of the dosage form depends on the method of administration and can be easily determined by a person skilled in the art. In some embodiments, the dosage form is sterile or sterilizable. In particular, the materials described herein are often sterile when pretreated with radiation exposure as described herein.

In some embodiments, dosage forms may contain carriers or excipients, many of which are known to those skilled in the art. Illustrative excipients that can be used include buffers (eg, citrate buffer, phosphate buffer, acetate buffer, and bicarbonate buffer), amino acids, urea, alcohols, ascorbic acid, phospholipids, polypeptides (eg, serum albumin), EDTO, sodium chloride, liposomes , mannitol, sorbitol, water and glycerin. Dosage forms can be made to be suitable for any standard route of administration. For example, administration can be parenteral, intravenous, subcutaneous, or oral, or it can be by any route of administration approved by REedegai Ogyd Aatipivzigayop (see mebr-address on the world wide web

Ttda.dom/sdaeg/dazt/Yu A s/ag900301. sweat).

Zo In addition to the compositions described above, the composition can also be made as a depot drug. Such long-acting compositions can be administered, for example, by implantation (eg, subcutaneously). Thus, for example, the compositions can be made with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, such as a sparingly soluble salt.

Pharmaceutical compositions prepared for systemic oral administration may be in the form of tablets or capsules prepared by conventional methods with pharmaceutically acceptable excipients such as binders (eg, pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose); fillers (for example, lactose, microcrystalline cellulose or calcium phosphate); lubricants (for example, magnesium stearate, talc or silicon dioxide); disintegrants (eg, potato starch or sodium starch glycolate) or wetting agents (eg, sodium lauryl sulfate). Many of the functions of these binders, fillers, lubricants, and disintegrants can be performed by the pretreated materials described herein.

Tablets can be coated by methods well known in the art. Liquid preparations for oral administration may be in the form of, for example, solutions, syrups, or suspensions, or they may be presented as a dry product for dilution with water or another suitable vehicle prior to administration. Such liquid preparations can be prepared by conventional methods with pharmaceutically acceptable additives such as suspending agents (for example, sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifiers (for example, lecithin or gum arabic); non-aqueous carriers (eg, almond oil, oil esters, ethyl alcohol or fractionated vegetable oils) and preservatives (eg, methyl or propyl-p-hydroxybenzoates or sorbic acid). Also, preparations may contain buffer salts, flavor additives, dyes and sweeteners, depending on the situation.

Preparations for oral administration can be suitably formulated to provide controlled release of the active compound.

In some embodiments, the materials described herein may be used as pharmaceutical ingredients for use in a topical iontophoresis, phonophoresis, fast-dissolving tablet, lyophilized film, intravaginal drug delivery system, vaginal liner, urethral liner or suppository, implantable pump for drug delivery, an external drug delivery pump and a liposome.

Hydrogels

In some embodiments, the materials described herein can be used in the manufacture of hydrogels. Hydrogels are three-dimensional networks of hydrophilic polymer chains that are cross-linked either by chemical or physical bonds, are insoluble in water, and are typically superabsorbent (eg, may contain more than 99 95 water) and allow gas exchange and exchange of nutrients.

In some embodiments, the materials described herein can be used to form a hydrogel. For example, to obtain a hydrogel, you can use monosaccharides, oligosaccharides and polysaccharides contained in the materials described in this document. Alternatively or additionally, the materials described herein can be used to form a hydrogel in combination with other materials such as hyaluronan, gelatin, cellulose, silicone, and one or more extracellular matrix (ECM) components.

In some embodiments, hydrogels containing the materials described herein may be cross-linked (eg, chemically cross-linked) and/or oxidized.

Alternatively or additionally, hydrogels containing the materials described herein may be cross-linked using low-intensity irradiation. Doses of low-intensity low-level radiation that can be used to cross-link the materials described herein include, but are not limited to, for example, 0.1 to 10 Mrad. Alternatively or additionally, hydrogels containing the materials described herein may be cross-linked using a combination of chemical cross-linking, low-intensity irradiation, and oxidation.

In some embodiments, the methods described herein can be used to modify (eg, increase) the average molecular weight of the biomass materials described herein. For example, the methods described herein can be used to increase the average molecular weight of biomass material, e.g.

From 10, 25, 50, 75, 100, 150, 200, 300 95 or up to 500 95.

In some embodiments, the methods described herein can be used to modify (increase or decrease) the Poisson's ratio of the hydrogel.

In some embodiments, hydrogels obtained using the materials described herein may include one or more biological cells and/or one or more bioactive agents, such as a pharmaceutical agent or ECM component. Candidate pharmaceutical agents may include, but are not limited to, a therapeutic antibody, an analgesic, an anesthetic, an antiviral agent, an anti-inflammatory agent, an RNA that mediates

RNA interference, microRNA, aptamer, peptide or peptidomimetic, immunosuppressant, hydroxyapatite or bioglass.

Hydrogels containing the materials described herein can be used as biodegradable or non-biodegradable implantable (e.g., subcutaneously implanted) three-dimensional scaffolds, e.g., in wound healing and tissue engineering, implantable disc prostheses, drug delivery carriers (e.g., carriers for delivery of sustained-release drugs), in wound dressings, contact lenses, and as superabsorbent materials (e.g., in diapers).

Hydrogels containing the materials described herein can also be combined with medical devices to treat both external and internal wounds. Hydrogels can be applied to wound dressings, such as non-healing chronic wounds, or used as subcutaneous implants. Alternatively, these hydrogels can be used in organ transplantation, such as liver transplantation from a living donor, to stimulate tissue regeneration. Hydrogels can be tailored to specific tissue types by balancing water content, biodegradation kinetics, and Poisson's ratio with these parameters of the target tissue to be repaired.

Methods of obtaining hydrogels are well known in the art and can be found, for example, in 0.5. 2006/0276608.

Absorbent materials

In some embodiments, the methods described herein can be used to prepare absorbent materials. For example, in some embodiments, biomass can be processed using one or more of the pretreatment methods described in this document. Such materials may have, for example, a modified

(increased, decreased, maintained) solubility, porosity, surface area, average molecular weight, functionalization (for example, increased number of hydrophilic groups).

Alternatively or additionally, these materials can be chemically treated to enhance a particular absorption property. For example, materials can be treated with silanes to render them lipophilic. These materials can have the ability to absorb 1, 2, 5, 10, 20, 50, 100, 500 and 1000 times more liquid than the original materials and/or 1, 2, 5, 10, 20, 50, 100, 500 1000 times more than the own mass of materials. In some embodiments, these materials can be used to adhere (eg, selectively) one or more materials (eg, biological materials in blood or plasma, toxins, pollutants, waste materials, inorganic chemicals, and organic chemicals), for example, in solution or in a dry environment.

In some embodiments, the materials described herein can be used as absorbent materials, for example, for use as bedding for animals, such as small and large animals, and bedding for animals. Methods of making animal bedding are well known in the art (see, for example, US Pat. No. 5,352,780).

In some embodiments, the absorbent animal bedding further includes a scented or scented material and/or an odor-eliminating material as known in the art.

In some embodiments, the materials described herein can be used to absorb spilled chemicals, for example, by applying the material to the spilled substance.

In some embodiments, the materials described herein can be used in combination with a filter, such as a medical filter or a non-medical filter.

The materials described herein may provide suitable absorbent materials due to the high surface area, high absorbency, high swelling properties, and high porosity of the materials described herein.

Fight against pollution

In some embodiments, the absorbent materials described herein can

To be used to fight pollution. When absorbent materials are used for such applications, they can be used in solid, liquid, or gas form.

For example, the materials described herein can be used to absorb oil and/or to clean up contaminated environments, such as in water, in air, and/or on land. The materials described in this document can also be used for the treatment of waste water (eg industrial waste and sewage) and for water treatment.

In some embodiments, the absorbent materials described herein can be used in combination with biological agents (microorganisms, fungi, green plants, or enzymes thereof) or chemicals to aid in the removal, inactivation, or neutralization of a pollutant from the environment, such as using a biological restoration.

In some embodiments, the absorbent materials described herein may be degradable (eg, biologically degradable). Such a process can be controlled to achieve the desired rate of degradation. In some embodiments, the absorbent materials described herein may be resistant to degradation.

In some cases, these absorbent materials may be associated with a structure or carrier such as a net, membrane, floating device, bag, shell, filter, case, or biodegradable substance. Optionally, the structure or carrier itself may be made of the materials described herein.

Air purification

In some embodiments, the biomass processed using the methods described herein may carry a charge (eg, positive or negative) or it may be neutral. In some embodiments, charged (eg, positively or negatively charged) materials can be used to remove pollutants (eg, microorganisms, spores, mold spores, dust, pollen, allergens, soot particles, and dust mite residues) from the air. In some embodiments, charged (eg, positively or negatively charged) materials can be used to trap contaminants. Alternatively or additionally, charged (eg, positively or negatively charged) materials can be used to remove polluting agents. 60 For example, in some embodiments, the methods described herein can be used to increase the cation level of a material. As a rule, cationic compounds have antimicrobial activity. In some cases, charged (eg, positively or negatively charged) materials can be combined with phenolic resins, pharmaceuticals, and/or toxins (eg, those listed herein) to eliminate microorganisms and/or spores.

In some embodiments, charged (eg, positively or negatively charged) materials can be used with a device, such as an air purifier.

For example, charged (eg, positively or negatively charged) materials can be immobilized on a surface in an air cleaning device, such as a filter (eg, a fibrous filter and/or a fibrous filter in the form of a layer). Alternatively or additionally, charged (eg, positively or negatively charged) materials may be present in the air cleaning device in the form of gas and/or vapor. Alternatively or additionally, charged (e.g., positively or negatively charged) materials can be used in an air handling system (e.g., an air conditioner), such as an enclosed environment such as a vehicle (e.g., a car, bus, airplane, and train car). ), in a room, office or building. For example, charged (eg, positively or negatively charged) materials can be immobilized on a surface in an air handling system, such as a filter. Alternatively or additionally, charged (eg, positively or negatively charged) materials may be present in the air treatment system in the form of gas and/or vapor. Alternatively or additionally, charged (eg, positively or negatively charged) materials can be used more locally. In such cases, charged (eg, positively or negatively charged) materials may be contained in a container and dispensed from the container, such as a pressurized canister or a non-pressurized container, by a pump. Alternatively or additionally, charged (eg, positively or negatively charged) materials can be used in a sustained release system, for example, where the charged (eg, positively or negatively charged) materials are released into the air over a period of time. Such sustained release systems are known in the art and are commercially available. In some embodiments, the implementation in such systems with a delayed release to provide release

Charged (eg, positively or negatively charged) materials can be used to generate heat (eg, generated using electricity).

In some embodiments, charged (eg, positively or negatively charged) materials can be used with the air filter.

In some embodiments, charged (eg, positively or negatively charged) materials can be used in a device designed to filter air that is inhaled and/or exhaled by a person (eg, masks, filter helmets, and/or filter suits). In some embodiments, such devices can be used to reduce human inhalation of one or more potential pollutant agents.

Alternatively or additionally, such devices can be used to reduce human exhalation of one or more potential polluting agents.

In some embodiments, the methods described herein can be used to produce materials suitable as flavoring agents. Such flavors can be combined with any of the products and co-products described in this document.

Alternatively or additionally, flavoring agents may be used to alter the odor or flavor of a material (eg, solid or liquid) and/or air. In such cases, fragrances can be used in combination, for example, with candles, perfumes, detergents, soaps, gels, sprays and air fresheners. Illustrative flavors that can be derived from biomass include, for example, lignin and bioflavors.

Food preservation

In some embodiments, the methods described herein can be used to produce materials that are suitable for food preservation or that can be used for food preservation. In such cases, suitable materials can be in the form of gas, vapor, liquid and/or solid. In some embodiments, materials (eg, charged materials) can be used to trap contaminants. Alternatively or additionally, materials (eg, charged materials) can be used to remove fouling agents. In some cases, materials (eg charged materials) can be combined with phenolic resins and/or toxins to eliminate microorganisms and/or spores. For example, materials (e.g., charged materials) can be used to remove contaminating agents (e.g., microorganisms, spores, and mold spores) from the environment surrounding food components to prevent,

limiting or reducing spoilage of food components. For example, materials (e.g., charged materials) may reside in a container in which food components are transported. Alternatively or additionally, the materials (eg, charged materials) may be contained in a container (eg, a package or bag) designed to store the food component. Such components may be sold with materials (eg, charged materials) that may already be present in them, or materials (eg, charged materials) may be added when the food component is added to the container. Alternatively or additionally, the materials (eg, the charged materials) may reside in refrigerated storage, such as a refrigerator and/or freezer.

Herbicides and pesticides

In some embodiments, the methods described herein can be used to produce toxins (eg, natural toxins), including, but not limited to, herbicides and pesticides. Such materials include, for example, lectins, glycoalkaloids, patulin, algal toxins, paralytic shellfish poison (PP), amnesic shellfish poison (A5P), diarrheal shellfish poisons (O5P), vitamin A, and mycotoxins.

Fertilizer

In some embodiments, the methods described herein can be used to produce materials that can be used as fertilizer. Biomass is rich in nutrients and is currently used as a fertilizer, but the raw material has low solubility and is suitable as a fertilizer only after partial or complete decomposition processes, both of which take a significant amount of time, require some monitoring and require the provision of storage space for the time while decomposition takes place. This mainly limits the use of biomass as fertilizer.

In some embodiments, the methods described herein can be used to modify biomass into materials, for example, with modified (eg, increased) solubility, which can be used as fertilizers. Such materials can be spread over an area requiring fertilizer and will solubilize when in contact with a solution (such as water and rainwater). This solubilization can make the nutrients in the materials more available to the area that needs the fertilizer.

In some embodiments, the methods described herein can be used to modify biomass into materials for use as fertilizers. Such materials can be combined (eg, mixed) with seed, nitrate, nitrite, nitrogen, phosphorus, potassium, calcium, lime, vitamins, minerals, pesticides, and any combination thereof.

Alternatively or additionally, such materials can be combined with one or more microorganisms capable of degrading the materials and/or one or more enzymes capable of degrading the materials. These components can be provided together or individually in liquid or dry form. In some cases, these materials may be associated with a structure or carrier, such as a net, membrane, floating device, bag, shell, filter, case, or biodegradable substance. Optionally, the structure or carrier itself may be made of the materials described herein. In some embodiments, these materials and combinations of these materials can be mixed in a container (eg, bag or solid container), for example, to ensure decomposition. Such mixtures may be provided for use in a container (eg, a bag or rigid container).

In some embodiments, the methods described herein can be used to produce materials that can be combined with plant seeds. For example, the materials obtained using the method described in this document can be applied to the surface of the seed, for example, to protect the seed from decay, to protect the seed from microorganisms and/or to fertilize the seed.

Chemical and biological applications

In some embodiments, the methods described herein can be used to prepare materials suitable for use as acids, bases, and/or buffers. Such materials can be used, for example, to change and/or buffer the pH of a material (eg, solid or liquid) that requires such treatment. Such materials include non-edible solids and liquids and/or edible solids and liquids (for example, foodstuffs such as meat products, beverages and dairy products).

In some embodiments, the methods described herein can be used to prepare materials suitable for use in supporting or stimulating the growth of 60 microorganisms (e.g., bacteria, yeasts, fungi, single-celled organisms such as algae, protozoa or fungi-like single-celled organisms, e.g. slime mold) and/or plants and trees.

Lignin

In some embodiments, the methods described herein can also be used to produce lignin, such as lignin residue.

Lignin is a phenolic polymer that is usually associated with cellulose in biomass, such as plants. In some cases, in the methods described in this document, lignin is formed, which can be obtained (for example, isolated or purified) from the biomass feedstock described in this description. In some embodiments, the lignin produced by any of the processes described herein can be used, for example, as a plasticizer, an antioxidant, in a composite (e.g., a fiber-resin composite), as a filler, as a reinforcing material, and in any which of the pharmaceutical compositions described in this document.

In addition, as described above, lignin-containing residues from the main and pretreatment processes have value as high/medium energy fuels and can be used to generate power and steam for use in manufacturing processes. However, such lignin residues represent a new type of solid fuel and the off-site demand for it may be low, and the cost of drying it for transport may reduce its potential value. In some cases, gasification of lignin residues can be used to convert it into a high-value product at a lower cost.

In some embodiments, lignin can be combined with one or more of the products or co-products described herein. For example, lignin can be combined with one or more herbicides and/or pesticides, for example, to produce a delayed release system, for example, where one or more herbicides and/or pesticides are released over a period of time. Such sustained release systems can be combined with the fertilizers described in this document. Alternatively or additionally, lignin can be combined with charged (eg, positively or negatively charged) materials to produce a sustained release air cleaning system. In some embodiments, lignin can be used, for example, alone or in combination with one or more of the products and co-products described herein as a composite,

For example, for use as an additive in plastics and/or resins. An example of the structure of lignin is presented below.

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Other products

Cell material, furfural and acetic acid have been identified as potential co-products of biomass fuel plants. Interstitial cellular material can be valuable, but may require significant purification. Sales markets for furfural and acetic acid are relevant.

Products of bioconversion

As described above, the methods described herein can be used to process biomass to obtain/produce, for example, food products (e.g. food for animals (including aquatic animals), humans and/or microorganisms), proteins, fats and oils , carbohydrates and sugars, vitamins, minerals, ash, pharmaceuticals, nutraceuticals and nutraceuticals, pharmaceutical dosage forms, hydrogels, absorbent materials, air purification materials, food preservatives, herbicides and pesticides, fertilizers, acids, bases and buffers, and lignin. As shown in Fig. 43A, these methods typically include processing the biomass, such as altering (eg, reducing) the recalcitrance level of the biomass, to produce products, such as those derived directly from the biomass, and/or to produce products containing these materials.

Alternatively or additionally, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to yield a second material that can be used as a substrate for additional processes, e.g. to obtain materials and products present (eg, substantially present) or in excess in the first material In some embodiments, additional processes may include a bioconversion step, as shown in FIG. 43B. In some embodiments, the bioconversion step may include the use of microorganisms. Examples of methods that include a bioconversion step are described above, for example, when applying the methods described herein for the production of energy products (e.g., ethanol), alcohols and/or organic acids, all of which are optionally present (e.g., essentially absent) or present in excessive amounts in natural unprocessed biomass. Additional examples of such methods are described below.

Food

In some embodiments, the methods described herein can be performed in combination with a bioconversion step (eg, see FIG. 43B) to produce a food product (eg, an ingestible product such as a food product, such as a food starch and/ or protein) for animal or human use. One of the advantages of such methods over conventional methods of agricultural food production is that the methods described herein do not require large areas of land and can be carried out in environmental conditions that are not favorable for conventional production methods.

Malnutrition, particularly protein-calorie deficiency, is a growing problem worldwide, particularly in developing countries. Insufficient calories and protein lead to an increase in infectious diseases, stunting of physical growth, and slowing of brain and mental development. These problems of undernutrition are caused by growing populations around the world, combined with inadequate food supplies in developing countries and aging food production methods. Without changes in population growth, food supply and production methods, malnutrition will also soon become a serious problem in developed countries.

One of the solutions to these problems is to increase the food supply. However, this may be difficult under conventional agricultural practices due to the limited availability of land for agronomy and documented global climate change. In addition, conventional agricultural practices are not favorable in certain environmental conditions, such as environmental conditions in which excessive heat or limited oxygen and/or limited sunlight are present. An alternative solution is to modify the usage

From currently available materials (eg biomass) to create alternative food sources, for example to increase the nutritional value or suitability of already available materials.

The use of microbial proteins as a food product for animal and human consumption is known in this field and is monitored by Te Rosa apa Adgisiiyge Ogdapigayop oi She

Opyeia Maiop5 (BA). BAO, in collaboration with the World Health Organization (WHO), has published publicly available reports outlining the guidance and standards required for food products produced by biotechnology (see, for example, “oipi

EAOLINO Ekhrep SopziyMayop op Roodv Oegimed iot Vioyespppoiodu, 1996; 5ieme Tauyiog, doipi

EAOLUNO Ekhrep Sopzyinaiyop op Rosaz Oegimed iot Vioiespoiodu, 2001 (Vioiiesp 01/03); to you

Om, ChdChoipi EAOLINO Ehrepy SopzyMayop op EROode Oegimei iot Vioyesnpoiodu, 2000 (Viotesn 00/14)3). This guidance outlines the safety issues that must be considered when using microorganisms for food production, the types of organisms that are suitable for such use, and the requirements for the proteins produced (see, for example, Sotitivviop oi

Sepeiis Vezotsgsev Tog Ros apa Adgisiyete, 1118 bezvziop, Vote Uipe 11-15, 2007, published reference SOKEA-11/07/Sigs.3).

The use of microorganisms and microbial proteins as a food source is justified by their known long-term use as food. For example, the Indonesian plant tempeh is combined with the fungus (for example, a mold) Kpi2oryh oiidozorogy5 and consumed. Algae are also used as a food source in populations along the shores of Lake Chad and Lake Texcoco in Mexico City, and currently in

Mexico City produces thousands of tons of spirulina as a protein-rich food source. In the mid-1960s, a quarter of a million tons of nutritional yeast were produced, and by 1970, the Soviet Union planned an annual production of 900,000 tons of nutritional yeast to compensate for the shortage of agricultural protein (VipKeg, "Mem/ Rosa", 2pa Ipi Sopdg. Rooy osi. apa Tesppo). ,

UmMagzam, p. 48 (1966)). As a result of marked improvements in crop production, increased communication between countries with food surpluses and shortages, and increased oil prices, microbial protein production has not developed as predicted. However, the protein obtained from Rizagisht mepepaisht is currently approved for use in Europe and is sold in the USA under the trade name OtsogpF (for review see MMebre, Musoipodivy, 18:17-20, 2004).

The use of microbial proteins as a food source for animals and humans further confirmed the 60 observation that the chemical composition and levels of microbial protein from bacteria, fungi (e.g.

yeast and mold) and algae are comparable to these indicators in soybean meal. Moreover, the amino acid composition and digestibility (including total energy (kcal/kg), based on pig data) of microbial proteins from yeast, bacteria, fungi, and algae have also been reported to be comparable to those in soybean meal (see, for example, Khoipo et al., US patent Mo 4938972).

In some embodiments, the food products described below can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Squirrels

Methods of obtaining microbial proteins using cellulosic materials are described in the art (see, for example, Catazatu ei ai., 9U. Arri. Viotespoi!., 46:117-124, 1979, Moshpa ei a!., Vioieispoi I ei, 14 :863-868, 1992, Apirata apa Ramipaga, Vgaliyap Agspime5 og Vioiodu apa

VioyesNnpoi. 44:79-88, 2001, US Patents MoMo 3627095, 4379844, 4447530, 4401680, 4526721, 5047332 and 4938972).

In some embodiments, the methods described herein can be performed in combination with a bioconversion step (eg, see Fig. 43B) for protein production. In some embodiments, a second material is used as a substrate for microorganisms that converts the organic material present in the second material into proteins, such as microbial proteins (for example, when combined with a nitrogen source). In some embodiments, proteins can be used as or in products for ingestion (eg, food) for animal and/or human consumption.

The term "microbial proteins" includes unicellular proteins (UC), a term coined in the 1960s to encompass microbial biomass produced by fermentation, in which the microbial cells are typically released from the substrate, and microbial biomass products (MBPs), material in which the substrate is not cleaned of ZSR.

Illustrative microbial proteins can be obtained from cells of bacteria, fungi (eg, yeast and mold), and/or algae. With proper cultivation, these cells can contain more than 40 95 protein per dry weight. One of the advantages of using microbial proteins as a potential food source is that microbial protein is easily renewable and readily available.

With a resource. For example, 1,000 kg of yeast can produce 12,000 kg of new cells containing 6,000 kg of protein in 24 hours.

In some embodiments, microbial proteins can be produced using the methods described herein to process a first material (e.g., biomass) into a second material (e.g., substrate) that is provided by one or more of bacteria, fungi (e.g., yeast and mold ) and/or algae, for example, in the presence of nitrogen or a source of nitrogen, in the presence or absence of oxygen and at the temperature and pH required by the organism or mixture of organisms for protein synthesis (for example, at a level above the normal level of protein synthesis in the cell). As a rule, these methods include the use of any microorganism that synthesizes protein in the presence of materials obtained using the methods described in this document. Such organisms are generally suitable, or can be made suitable, for animal and/or human consumption. In some embodiments, the microorganism may be a non-pathogenic organism and/or an organism generally recognized as safe (OKA5B). Additional selection criteria considered when selecting a microorganism may include, for example, whether the organism is capable of producing large amounts of proteins (eg, food proteins or proteins that can be converted into food proteins) or whether it can be modified to produce them ; whether isolated cultures of the organism are commercially available and whether the organism can be effectively isolated; can the microorganism be easily maintained in culture; whether the microorganism is genetically stable; and whether the organism can effectively dispose of the substrates produced using the methods described herein (eg, whether the microorganism can be cultured on the substrate provided).

In some embodiments, microorganisms can be modified (eg, by engineering methods) to express one or more recombinant proteins, such as proteins that are not normally encoded by microorganisms. For example, these proteins may be proteins known to have high nutritional value for humans and/or animals (e.g., as determined by evaluating the biological value (BI) of the protein (e.g., fraction of residual nitrogen absorbed) and/ or the use of pure (MP) protein (eg fraction of residual protein used)). In experimental animals, MRO can be directly estimated by cadaver analysis, and the values are thus likely to be more accurate than the VM and MRI derived from M balance data in human studies. The inaccuracy inherent in 60 studies of the M balance is known, and it does not matter how carefully it is carried out. Thus, MRI and VM are indicators of the same parameter (residual M, except that

VM is calculated from absorbed M, and MRI is calculated from used M (for a review, see, for example, Vepaeg,

Veiayop Veimeep Rgoivip EPisiepsu apa Mei Rgoivip ShNii2ayop, Meazigetepi oi Rgoivip Shii2ainop, 10:135-143, 1956)). In some embodiments, proteins with high nutritional value may have a high BM at the applied level (mg/kg) necessary to meet the recommended daily protein requirements of an animal and/or human, and may contain suitable levels of essential amino acids (EAAs) required for the formation of protein in the animal or human body (EAAs include, for example, phenylalanine (the GAO recommended daily intake is 2.2 g); methionine (the RADO recommended daily intake is 2.2 g); leucine (the ECHO recommended daily intake is 2.2 g ); valine (the recommended EAO daily intake is 1.6 g); lysine (the EGAO recommended daily intake is 1.6 g); isoleucine (the ECHO recommended daily intake is 1.4 g); threonine (the EAO recommended daily intake is 1, 0 g) Its tryptophan (the daily intake recommended by RADO is 0.5 g)). In some embodiments, the high nutritional value proteins may be synthetic proteins, e.g., designed to have a high BM at consumption levels necessary to meet the recommended daily requirements of an animal and/or human, and may contain suitable levels of all EAA, necessary for the formation of protein in the body of an animal or a person. In some embodiments, proteins with high nutritional value can be labeled (eg, labeled), for example, to facilitate protein identification and/or purification. Such proteins are also referred to herein as microbial proteins.

Illustrative fungi that can be used in the methods described herein include, but are not limited to, Azregoii5 piedeg, A. lipidagi5, A. ieggei5, SospiioboiIi5 vresiiteg, Mugoipesiitis metisaipa, Vpigosiupia 5oiiiapi, Zrisapa hyvivroga, Repisyit vr., Siiosiadiitis 5r ., Eryzapytst 5 years.

Eizagitst mepepaisht (formerly R. dgatipeagyt) strain A 3/5 (for example, ATSS 20334). Suitable cultivation conditions for this organism are described in US Plant Patent Mo 4347 and Pat

of Europe Mo 123434. R. 501 Iapi, RE. ohuzrogiit and Raesiotusez magioiiii, mycelium, Kpigory5 oiYidozrogiv,

Sapaida schi5 ii Zasspagotusez segemiziae. Illustrative algae that can be used in the methods described herein include, but are not limited to, 5Zrigiiipa 5r.,

From Zsepedebzty5 asshiv5, Zrigiypa Tahita and So5tagisht ishgripii. Illustrative bacteria that can be used in the methods described herein include, but are not limited to, KpodozrigPyt 5r., and Kpodorzendotopazh 5r., Soguperasiegiit dishatisit, E5spegisnpia soii,

AiIsaiiidepez Taesaiiii5, Tpegtotopozroga iTizsa (Asiiipotuseiiaseae) and Rhsendotopaz UM127.

In some embodiments, microbial proteins, such as BSR, can be provided for use in

C5 food by animals and/or humans, for example, without release of a microorganism or a mixture of microorganism. In such cases, cells containing 5SR can be concentrated using, for example, filtration, precipitation, coagulation, centrifugation and the use of semipermeable membranes. Cells containing ZSR can also be dried, for example, to a moisture content of about 10 95 and/or they can be compacted and acidified to limit spoilage.

In some embodiments, the SR can be provided for animal and/or human consumption shortly (eg, within 12 hours, 24 hours, 48 hours) after production without further processing of the 5SR. In some embodiments, SSRs can be used in the absence of additional food sources (see the EAO publication regarding the recommended daily intake of SSRs for animals and humans). Alternatively or additionally, 5SR can be combined, for example mixed, with other food sources before or simultaneously with animal and/or human consumption. To obtain ZSR mixtures, ZSR can be combined with dry and/or wet food sources. In some embodiments, mixtures containing ZSR can be processed, for example, as described by Tappeprast (US patent Mo 3925562). For example, 5CP microorganisms can be combined with an auxiliary protein (for example, vegetable protein) and structured into a paste suitable for use as a food additive. Such methods can be used to add 5CP properties to the desired texture.

In some embodiments, protein utilization and nitrogen assimilation from protein material

ZSR can be increased by homogenization of cells (see, for example, Uapod eyi ai., 9. Boba 5soi., 42:1247-1250, 2006). Thus, in some embodiments, microbial proteins can be extracted or isolated from a microorganism or a mixture of microorganisms prior to consumption by animals and/or humans. For example, microbial whites can be extracted by chemical, enzymatic and/or mechanical disruption of the microbial cell wall and/or membrane, for example, to release the intracellular contents of the cells. Microbial proteins can then be isolated or purified from contaminating materials using protein isolation methods,

known in the field. In some embodiments, microbial proteins can be isolated or purified using a detectable label fused to the protein.

In some embodiments, the microbial protein can be modified, for example, glycosylated and/or folded before use, for example, to make them more or less antigenic.

In some embodiments, microbial proteins can be isolated and hydrolyzed to individual amino acids, peptides and/or polypeptides, for example, prior to consumption by animals and/or humans.

Methods of protein hydrolysis are known in this field.

In some embodiments, the microbial bilayer can be purified (to at least 50 95 , such as 60, 70, 80, 90, 95, 99, or 100 95 w/w, w/v, or v/v) and not must concentrate. The protein structure can then be modified to resemble the fibrous structure of animal muscle proteins before the product is flavored with meat using meat flavorings and fats. In some embodiments, microbial proteins can be used as the primary source of protein in a meat analog. Alternatively, microbial proteins can be used to supplement currently available meat analogs, such as meat analogs sold under the trade name OiogpF) and soy protein-based products.

Fats, oils, lipids and hydrocarbons

In some embodiments, the methods described herein can be performed in combination with a bioconversion step (eg, see Fig. 438) to produce fats and/or oils.

The market for fats and oils is large and extremely diverse, ranging from mass goods used for food and technical purposes to more specialized oils.

The use of microbial fats and oils is known in the art (for a review on this topic, see, for example, Rgude, Mem/ 5o!tsgsez oi Raiv apa OiiYi5, Ateg OIII Spetivi5 5osieiu, (Ategisap OIII Spetivi

Bossiviu (ADOS5), 1981)).

In some embodiments, fats and/or oils obtained using the methods described in this document can be used, for example, as substitutes for animal and vegetable fats and oils, in the production of energy products, combustible substances (solid and/or liquid), in the production and preparation of food, as flavor enhancers (for example, for food

From products) as animal feed or in animal feed as food additives or in food additives as pharmaceuticals or in pharmaceuticals as nutraceuticals or in nutraceuticals as cosmetics or in cosmetic products and as nutritional therapy after surgery or in nutritional therapy after surgery.

In some embodiments, microbial fats and/or oils can be produced using methods described herein to process a first material (e.g., biomass) into a second material (e.g., substrate) that is provided to one or more of bacteria, fungi (e.g. , yeasts and moulds) and/or algae, for example in the presence of nitrogen or a source of nitrogen, in the presence or absence of oxygen and at the temperature and pH required by the organism or mixture of organisms for the synthesis of fats and/or oils (e.g. above normal level of synthesis of fats and/or oils in the cell). As a rule, these methods include the use of any microorganism that synthesizes fats and/or oils in the presence of materials obtained using the methods described in this document. In some embodiments, the microorganism may be a non-pathogenic organism and/or an organism generally recognized as safe (OKA5B). Additional selection criteria considered in selecting a microorganism may include, for example, consideration of whether the organism is capable of producing large amounts of fats and/or oils or whether it can be modified to produce them; whether isolated cultures of the organism are commercially available and/or whether the organism can be efficiently isolated; can the microorganism be easily maintained in culture; whether the microorganism is genetically stable; and whether the organism can efficiently dispose of the substrates produced using the methods described herein (eg, whether the microorganism can be cultured on the substrate provided).

In some embodiments, microorganisms that can be used in the methods described herein, for example, to obtain or produce microbial fats and/or oils, include, for example, bacteria (e.g., mycobacteria, corynebacteria, and norcadia), algae (e.g., SPiohorpuia (Siadorpoga hyrezigiv, Viiidipdia tipita, Epiegotogrna iptesiipaii5), RIiaeorpuia (Adagite sgibgosite, Absorpoint podbosite and Gatipagia aidynaga) and

Kpodorpuia (Roiuzirpopia Iapoza, raitagia raiYitaie, NaiIozassiop gatepiasesht and Rogrpuga

Ieisoziysie)), seaweed and sea grass, yeast (for example, Sapaida 107, Stuyusossiv5

Iegtisoyish5, Napsepshtia zayigpyv, Hyrotusez Irogtega, H. "agkeui, Kpodoiogitsia dgasiiy5, K. (ogshoidez and

Sapaida sigmaga) and molds (for example, Azregoyi5 piashiapv, A. iegei5, Rizagiit topoiyogte,

Mysog siksipePoide5, RepisiPit 5ripshosite, APygoriz 5r).

In some embodiments, prior to use, microbial fats and/or oils obtained using the methods described herein can be separated from, for example isolated from, microbial cells. Alternatively or additionally, microbial fats and oils obtained using the methods described herein can be used without separation from the microbial cells.

Some microorganisms can be used to produce hydrocarbons. For example, as discussed in the "Technical Level" section 0.5. 2008/0293060, the content of which is incorporated herein by reference, a variety of organisms such as bacteria, algae and plants can synthesize hydrocarbons, for example n-alkanes with different carbon chain lengths, as previously described (Oeppi5, M.MU. 5 KoianikKiadu , R. E. (1991) Agspime5 oi Biospetivigu apa Biorpuzisv 287, 268-275;

Kipvi, Y. b Zativye!v5, A.I. (2003) Rgoagezz ip Iiriy gezvagsi 42, 51-80; TiPtap, 9.A., Zeuroid, 5.4).,

Uigtepka, V.A., 5 Viotadiivii, (3.U. (1999) Ipsesi Biospetivigu apa toiiesshiag Bioiosdu 29, 481-514; Totarepe, T.2x. (1982) Ehrepepia 38, 1-4, each of which is included as link).

Illustrative species that synthesize hydrocarbons are presented in Table A and Table B, below.

Table A

Carbohydrate-producing prokaryotes

Siovigidi! Vadaema apa /ipygoma, 2004, Viospet (Mozsomu), mine. 69, o5tidiit rabvieshiapyt 427-428 oMoviyustivsogyt 77711111

Sosospiotivviarepeo 77711111

Table B

Carbohydrate-producing eukaryotes ivit zaymit 341-349

Rodi od Speezrogotsodi apa Koianikiau, 1988, 9. Vio!. Snet., my! 263, 2738-

Carbohydrates, sugars, biopolymers and polymer precursors

A large number of biopolymers, such as polysaccharides, polyesters, and polyamides, are naturally produced by microorganisms (for a review, see Mysgobia!

Vioroiuteg5 apa Roiuteg Rgesygzog5, Kept, ey, (Saisieg Asadetis Rgev55, 2009)). These biopolymers vary from viscous solutions to plastics and their physical properties depend on the composition and molecular weight of the polymer.

In some embodiments, the methods described herein can be performed in combination with a bioconversion step (eg, see Fig. 438) to produce carbohydrates, sugars, biopolymers, and polymer precursors. In some embodiments, the methods described herein can be used to process a first material (e.g., biomass) to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria), fungi (e.g., yeast and mold), and/or algae) capable of producing, for example, xanthan, alginate, cellulose, cyanophycin, poly(gamma-glutamic acid), levan, hyaluronic acid, organic acids, oligosaccharides and polysaccharides, and polyhydroxyalkanoates. Applications of such carbohydrates, sugars, biopolymers and polymer precursors include, for example, applications as food additives, in cosmetics, in the manufacture of plastics, in the manufacture of fabrics and in pharmaceuticals and nutraceuticals.

As a rule, these methods include the use of any microorganism that synthesizes one or more carbohydrates, sugars, biopolymers and/or polymer precursors in the presence of materials obtained using the methods described in this document. In some embodiments, these methods include the use of any microorganism that synthesizes one or more of xanthan, alginate, cellulose, cyanophycin, poly(gamma-glutamic acid), levan, hyaluronic acid, organic acids, oligosaccharides and polysaccharides, and polyhydroxyalkanoates in the presence of materials obtained using the methods described in this document. In some embodiments, suitable organisms are suitable, or can be modified to be suitable, for animal and/or human consumption, or they can be generally recognized as safe (KAB).

Additional selection criteria taken into account when selecting a microorganism may include, for example, whether the organism is capable of producing large amounts of carbohydrates, sugars, biopolymers and/or polymer precursors (e.g., xanthan, alginate, cellulose, cyanophycin, poly(gamma-glutamine acid), levan, hyaluronic acid, organic acids, oligosaccharides and polysaccharides, and polyhydroxyalkanoates) or whether it can be modified to produce them; whether isolated cultures of the organism are commercially available and/or whether the organism can be efficiently isolated; can the microorganism be easily maintained in culture; whether the microorganism is genetically stable; and whether the organism can effectively dispose of the substrates produced using the methods described herein (eg, whether the microorganism can be cultured on the substrate provided).

Vitamins

In some embodiments, the methods described herein can be carried out in combination with a bioconversion step (eg, see Fig. 43B) to produce vitamins, for example, including, but not limited to, vitamin riboflavin (vitamin B2), vitamin B12, and vitamin C

In some embodiments, the substrate is used by the microorganism AbPpBbua dozv5iguyiY, and the produced vitamin is riboflavin (vitamin B2).

In some embodiments, the substrate is used by the microorganisms Vasiyi5 tedaipegisht, Rzhendotopah depygiysap5 and/or species of the genus Phoriopyrasiegisht, and the vitamin produced is vitamin B12.

In some embodiments, the substrate is used by the microorganism Zasspagotuse5 5r., and the produced vitamin is vitamin C.

In some embodiments, vitamin products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Edible mushrooms

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for cultivating or growing edible mushrooms These fungi can be used as a higher quality food source than a first material (eg biomass) and a second material that can be consumed by animals and/or humans as food.

Edible mushrooms are mushrooms that grow above ground on a suitable source of nutrients. As used herein, the term "edible mushroom" refers to edible mushrooms, including, but not limited to, mushrooms with a stem (stump), a cap (umbrella), and a spore-bearing layer (plate) on the underside of the cap, and mushrooms without peduncles, fresh fruiting bodies of some Absotusoia, woody or leathery fruiting bodies of some

Vasidiotusoia, and spores of edible mushrooms. In some embodiments, the term edible mushroom includes mushrooms that are edible to animals.

In some embodiments, suitable edible fungi of the present invention include, but are not limited to, for example, mushrooms, fungal mycelium, and fungal spores of Rieslings,

Vasidiotusoia, Adagisoptuseii5, MyimageNPa moiYimasea (radius mushroom), Rieigoii5 o5igeaiyi5 (common mushroom), Adagisib5 bBivrogib5, BiIattiiiypa meishireb5, Riecygoii5 egupadia, mushrooms Sapodepta and

Sohauserv.

Methods of cultivating edible mushrooms are known in the art (see, for example, US patent No. 6,737,065). After cultivation, edible mushrooms can be collected and stored for later

From the application or they can be used immediately. Mushrooms have a relatively low protein content (eg 2-5 95) on a raw weight basis, however, the protein content of edible mushrooms can be increased by drying the edible mushrooms (eg 30-50 95 on a dry weight basis). Thus, in some embodiments, edible mushrooms obtained using the methods described herein may be dried (eg, lyophilized) or dehydrated prior to use, such as ingestion. In some embodiments, edible mushrooms can be mixed with an auxiliary protein and binder and can be textured.

Growing plants without soil

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used in soilless plant cultivation. Growing plants without soil is a method of growing plants using solutions of mineral nutrients without soil. Plants can be grown with their roots only in a solution of mineral nutrients (culture in solution) or in an inert medium (culture in medium) such as perlite, gravel or mineral wool. The three main types of solution cultivation are static solution culture, constant flow solution culture, and aeroponics. The materials obtained using the methods described in this document can be used alone or in combination with microfertilizers, such as potassium nitrate, calcium nitrate, potassium phosphate and magnesium sulfate, to produce a hydroponic solution. Various micro-fertilizers can also be incorporated to provide essential elements such as Re (iron), Mn (manganese), Si (copper), P (zinc), B (boron), Si (chlorine) and Mi (nickel). Chelating agents can be added to increase iron solubility. During the life cycle of plants, various hydroponic solutions can be used to enhance growing conditions.

Aquaculture

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used in aquaculture. For example, the second material can be used for feeding or other maintenance of aquatic species. Aquaculture is the cultivation of freshwater and saltwater organisms, including molluscs, crustaceans and aquatic plants. Unlike fishing, aquaculture, also known as aquaculture, involves the cultivation of aquatic populations under controlled conditions. Mariculture refers to aquaculture carried out in the marine environment. Specific types of aquaculture include algae cultivation (growing brown algae/sea grass and other algae), fish farming, shrimp farming, oyster farming, and cultured pearl farming. Aquaponics combines fish breeding and plant breeding using symbiotic cultivation of plants and aquatic animals in a recirculating environment.

Production of food Rizagiit mepepash

In some embodiments, the methods described in this document can be used to process a first material (e.g., biomass), for example, to change (e.g., reduce) the level of recalcitrance of the biomass, to produce a second material that can be used as a substrate for the production of food Rizagisht mepepaisht (for example, which are supplied to the market under the trade name OptsogpF). Methods of production of OtsogpF are described, for example, in US patents MoMo 5935841, 6270816, 5980958 and 3809614, and reviewed in M/eire (M/eire, Musoiodivi, 18:17-20, 2004). In modern methods of production of OtsogpF, glucose is used as the main source of carbon. Substitution of glucose with the substrate described in this document can reduce the costs associated with the production of OtsogpF) because the substrates provided in this description provide a cheaper carbon source than glucose.

Alcohol

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for producing an alcohol that suitable for human consumption.

Such alcohols can be used as alcoholic beverages or in their products. For example, alcohols produced using the methods described herein can be used in the production of beer, wine, spirits and/or carbonated beverages containing alcohol.

Products for health

In some embodiments, the methods described herein may be used

For the processing of the first material (for example, biomass), for example, for changing (for example, reducing) the level of recalcitrance of the biomass, for the production of a second material that can be used as a substrate for obtaining health products for use in animals or humans. Such health products may include, for example, pharmaceuticals, nutraceuticals, cosmetics, medical cosmetics, and beauty products (eg, creams and lotions (eg, for application to the skin and/or hair)). In some embodiments, these health products may include, for example, functional foods that do not necessarily provide any nutritional value, but that increase gastrointestinal motility, or that can be used to lower cholesterol levels ( for example, high-fiber products that include soluble and/or insoluble fiber and products that contain soluble and/or insoluble fiber).

Amino acids and amino acid derivatives

Biotechnological methods have been used in the industrial production of amino acids for 50 years (for a recent review see I enispiepregdeg ei a!., Arri. Misgobioi. Viotesppoi!., 69:1-8, 2005).

Major products include flavor enhancers and animal feed products such as I-lysine, 1-threonine and I-tryptophan, which are usually produced using high-performance strains.

Soguperasiegisht dishatisit (see Kipospya ei ai., bep. Arri. Misgobioi., 3:193-205, 1957, and

Kailipomuv Ki ei ai., U. Violspoi., 104:5-25, 2003) and E5sPegispia soii, and substrates such as molasses, sucrose or glucose (I enspiepregeg, above).

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (for example, yeast or mold) and/or algae) capable of producing amino acids and/or amino acid derivatives (for example, when combined with a nitrogen source). These amino acids and derivatives can be used, for example, as flavor enhancers (for example, for food products), in animal feed, as food additives and in the production of pharmaceuticals, nutraceuticals, cosmetics and in post-operative nutritional therapy.

In some embodiments, amino acids and amino acid derivatives that can be expressed using the methods described herein include, but are not limited to, for example, I-amino acids and O-amino acids, such as Ii-glutamic acid (monosodium glutamate (M55 )), I -aspartic acid, I -phenylalanine, I -lysine, I-threonine, I-tryptophan, I1-valine, I-leucine, I -isoleucine, I -methionine, I-histidine and I-phenylalanine, I - lysine, and -methionine and 1- tryptophan.

For example, the aromatic amino acids tryptophan, phenylalanine, and tyrosine are biosynthesized from glucose via the shikimic acid cascade (shown below).

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The shikimic acid cascade converts simple carbohydrate precursors, formed during glycolysis and in the pentose phosphate cascade, into aromatic amino acids. One of the intermediate compounds of the cascade is shikimic acid, which gave its name to the entire sequence of reactions. The shikimic acid cascade exists in plants, fungi, and bacteria, but is not found in animals. Animals do not have ways of synthesizing the three aromatic amino acids - phenylalanine, tyrosine and tryptophan, which, therefore, are indispensable nutrients in animal diets.

In some embodiments, these amino acids can be modified to produce amino acid derivatives. Amino acid derivatives include, but are certainly not limited to, the following groups.

Amino alcohols nn on bzhryV, and АХ still and not t n MN";

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Gevnoletsnnoya Ietritofanog

Aminoaldehydes "a Zh Zh shi she ke

M-Vos-O-phenylalanine MA tert-butoxycarbonip-prod nal

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Muedymethyleneits Hydvoklornya M.sdymeraleinniahu Zhavyany etnlovnyy one

М.М-limetnaglitsnnu a 9 s: M tone "lita vash

In some embodiments, the microorganisms (e.g., bacteria, fungi (e.g., yeast and mold) and/or algae) suitable for use in the production of amino acids may be, but are not limited to, non-pathogenic organisms and/or organisms that are

SKA5. Additional selection criteria considered in selecting a microorganism may include, for example, consideration of whether the organism is capable of producing large quantities of a particular product or whether it can be modified to produce them; whether isolated cultures of the organism are commercially available and/or whether the organism can be efficiently isolated; can the microorganism be easily maintained in culture; whether the microorganism is genetically stable; and whether the organism can be cultured on the substrate provided. Alternatively or additionally, the microorganism may be a wild-type organism (eg, unmodified) or a genetically modified microorganism (eg, a mutant), for example, a microorganism that is modified or can be modified to overexpress one or more selected amino acids and/or amino acid derivatives. Illustrative microorganisms include, but are not limited to, lactic acid bacteria (LAB), E. soii, Vasilii5 5iIibBiib5, and Sogupebrasiegiit dishatisita (eg, ATCC 13032).

In some embodiments, amino acids and amino acid derivatives can be expressed using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution. In some embodiments, the methods and/or materials described in this document may be incorporated into methods currently used in Japan, Japan, AOM, USA,

Uedapud (South Korea), siora! VioSpet (China) and WABE apa Oedichza (Germany) for obtaining amino acids and amino acid derivatives.

Antibiotics

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate by microorganisms (e.g., bacteria, fungi (eg, yeast and mold) and/or algae) capable of producing antibiotics, for example, including but not limited to tetracycline, streptomycin, cyclohexamide, neomycin, cycloserine, erythromycin, kanamycin, lincomycin, nystatin, polymyxin B, bacitracin, daptomycin, vancomycin, and ansamycins or the natural products listed below.

Accurate brain protectors

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In some embodiments, the substrate is used by the microorganism 5igeriotusev5 geto5iv, and the produced antibiotic is a tetracycline.

In some embodiments, the substrate is used by the microorganism 5bigeriotuse5 dyzei5, and the produced antibiotic is streptomycin and/or cyclohexamide. The biosynthesis of streptomycin is illustrated below, starting with O-glucose.

Biosynthesis of streptomycin

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In some embodiments, the substrate is used by the microorganism Zigeriotuse5

Then, and the produced antibiotic is neomycin.

In some embodiments, the substrate is used by the microorganism 5bigeriotuse5 ogspidasetsv, and the produced antibiotic is cycloserine.

In some embodiments, the substrate is used by the microorganism 5bigeriotuse5 eguiphetsv5, and the produced antibiotic is erythromycin.

In some embodiments, the substrate is used by the microorganism 5bigeriotuse5

Kapatuseiisiv5, and the antibiotic produced is kanamycin.

In some embodiments, the substrate is used by the microorganism Zigeriotusev5

IpsoIpepv, and the produced antibiotic is lincomycin.

In some embodiments, the substrate is used by the microorganism Zigeriotuse5 poshgzei, and the produced antibiotic is nystatin.

In some embodiments, the substrate is used by the microorganism Vasi rotituha, and the antibiotic produced is polymyxin B.

In some embodiments, the substrate is used by the Vasiyya5 microorganism

Ispepitogitiv5, and the produced antibiotic is bacitracin.

In some embodiments, the substrate is used by the microorganism 5bigeriotuse5 gozeozrogiv, and the produced antibiotic is daptomycin.

In some embodiments, the substrate is used by the microorganism Atusoiiagrzib ogpiepiaiy5, and the produced antibiotic is vancomycin. The biosynthesis of vancomycin is described below, starting with POHIID GLUCOSE.

Biosynthesis of vancomycin by primary NAAS

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In some embodiments, the substrate is used by two strains of 5igeriotuse5 pudgossorisich, and the antibiotics produced are of the ansamycin family. The biosynthesis of 10 ansamycin is described below, starting from the glucose derivative.

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Carotenoids

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass) into a second material that can be used as a substrate by microorganisms (e.g., bacteria, fungi, yeast, mold, and/or algae), able to produce carotenoids, including, for example, β-carotene, lycopene and astaxanthin. Carotenoids are water-soluble natural pigments with 30-50 carbon atoms. Industrial uses of carotenoids include their use in nutritional supplements, for pharmaceutical purposes, as food dyes, and in animal feed.

Illustrative carotenoids in the automotive industry

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In some embodiments, antibiotic products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Vaccines

In some embodiments, vaccines are immunostimulating molecules (eg, low molecular weight compounds, peptides, and/or antigenic molecules). In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (e.g., yeast or mold) and or algae) capable of producing vaccines, including, for example, an influenza vaccine (e.g., a universal influenza vaccine, e.g., a universal influenza vaccine

MahIppaie Mav).

In some embodiments, vaccine products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Specialized chemicals

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (e.g. yeast or mold) and/or algae) capable of producing specialized chemicals, e.g. thickeners, xanthan (E 415), acidity regulators, citric acid (E 330), natamycin (E 235), nisin (E 234) and lysozyme (E 1105). In some embodiments, the methods described herein can be used to produce pure chemicals, such as flavors and aromas.

In some embodiments, chemicals can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Alcohols

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to modify (e.g.,

reduction) of the level of recalcitrance of biomass, for the production of a second material that can be used as a substrate for microorganisms (e.g. bacteria, fungi (e.g. yeast or mold) and/or algae) capable of producing alcohols, to supplement energy products (e.g. ethanol ), described above, for example, including, but not limited to, acetone and butanol. In some embodiments, the substrate is used by the microorganism Siovigidicht aseobshuyisit, and the alcohol produced is acetone. In some variants of implementation, the substrate is used by a mutant of the microorganism Siovigidiyt asyurshuiisite IPP 904 (ATOSS 39058), and the produced alcohols are acetone and butanol.

In some embodiments, alcohol products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Acids and bases

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (for example, yeast or mold) and/or algae) capable of producing acids and bases. In some embodiments, the substrate is used by microorganisms Aceiobrasieg and/or

Siisoporasieg, and the acid produced is acetic acid (for example, for use in the manufacture of vinegar).

In some embodiments, acidic and basic products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Enzymes

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (for example, yeast or mold) and/or algae) capable of producing enzymes.

Illustrative enzymes that can be produced using the methods described herein include, but are not limited to, for example, rennet enzymes, glucoamylase, polygalacturonase, cellulase, alpha-amylase, protease, beta-glucanase, pullulanase, amyloglucosidase, phospholipase, xylanase, monoglucose oxidase, novolipase, ultralipase, lipase, maltogenic amylase, alpha-acetodecarboxylase, soft protease, pectinesterase, carbohydrase, cellobiose oxidase, pectinlyase, monoxylanase, transferase, wheat xylanase, phytase, subtilisin, K-1-alpha-amylase, pectate , mannanase, trypsin and laccase.

Application of such enzymes (for example, individually or in combinations of one or more enzymes), for example in juice production, brewing industry, starch industry, bakery industry, oil and fat industry, meat industry, dairy industry, alcohol industry, animal feed industry animals, the detergent industry, the textile industry and the industry of personal hygiene products, known in this field.

In some embodiments, enzyme products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Growth factors

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (eg, yeast or mold) and/or algae) capable of producing growth factors.

Illustrative growth factors that can be obtained using the methods described herein include, but are not limited to, insulin-like growth factor, keratinocyte growth factor (KOR)-1 and -2, epidermal growth factor, fibroblast growth factor, granulocyte-macrophage colony-stimulating factor, human growth hormone, interleukin-1, platelet-derived growth factor, and transforming growth factor-B.

In some embodiments, growth factor products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution. because Plastics

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (eg yeast or mold) and/or algae) capable of producing plastics or plastic precursors. In some embodiments, the substrate is used by the microorganism

AlIsaidepe5 eshorpah, and the molecules produced are B-hydroxybutyrate and B-hydroxyvalerate.

In some embodiments, plastic products can be produced using a fed-batch fermentation process in which nutrients are added in a controlled manner according to the needs of the culture solution.

Fertilizers

In some embodiments, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g., bacteria, fungi (eg yeast or mould) and/or algae) capable of producing materials that can be used as or in fertilizers (eg proteins, fats and oils, carbohydrates and/or minerals). In some embodiments, the fertilizers produced using the methods described herein may be protein-based fertilizers or protein-enriched fertilizers (see Rapdio-ioppieppe et al., RMAB, 104:4524-4529, 2008, for a review of fertilizers based on proteins).

Methods of cultivation

As detailed above, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g. , bacteria, fungi (e.g. yeast or mold) and/or algae) to produce materials and products not necessarily present (e.g. essentially not present) or present in large quantities in the first

From the material. The choice of microorganisms will depend on the products produced.

Selection of microorganism

Several additional factors may also be considered when selecting suitable microorganisms for use in the methods described herein. For example, if the microorganisms are to be used to produce a health product for animal or human use, or if the microorganisms are to be used as food or in the production of food, the selected microorganisms are generally non-pathogenic and/or generally recognized as safe (OKAB). In addition, the selected microorganisms must be capable of producing large quantities of the desired product or they must be capable of being modified to produce large quantities of the desired product. In some embodiments, the microorganisms may also be commercially available and/or isolated efficiently, easily maintained in culture, genetically stable, and/or well characterized. The selected microorganisms can be wild-type microorganisms (for example, unmodified) or genetically modified microorganisms (for example, mutant microorganisms). In some embodiments, a genetically modified microorganism can be adapted to increase its production of a desired product and/or to increase the tolerance of the microorganism to one or more environmental factors and/or experimental conditions, for example, the microorganism can be modified (e.g., by engineering methods) to withstand temperature, pH, acids, bases, nitrogen and oxygen levels outside the range normally tolerated by the microorganism. Alternatively or additionally, microorganisms can be modified (eg, by means of engineering) to tolerate the presence of additional microorganisms. In some embodiments, microorganisms can be modified (eg, by engineering methods) to grow at a desired rate under desired conditions.

Culture solutions

As detailed above, the methods described herein can be used to process a first material (e.g., biomass), such as to alter (e.g., reduce) the recalcitrance level of the biomass, to produce a second material that can be used as a substrate for microorganisms (e.g. , bacteria, fungi (for example, yeast or mold) and/or algae), for example, in a culture solution. Generally, culture solutions can be prepared based on their ability to support the growth of 60 selected microorganisms. In addition to biomass-based substrates obtained as described herein, culture solutions may also optionally include an additional carbon source (eg, glucose), water, salts, amino acids, or a source of amino acids. In some embodiments, the culture solutions include an additional nitrogen source. The pH of these culture solutions can be adapted to the needs of the selected microorganism. Culture solutions may also optionally include one or more antibiotics to prevent contamination.

Some culture solutions are commercially available, for example, commercially available growth media include Hygia Vegiapi medium (IV), Iegitiis rgoii medium (TV), yeast and mold broth (UM) (yeast extract C g/L, malt extract C g/l, peptone 5 g/l and dextrose 10 g/l and pH 6.0-pH 8.0), URO medium (yeast extract, 3 g; mycological peptone, 5 g; U-glucose, 10 g per liter water) and bactopeptone. Growth media can be purchased from commercial sources (e.g. BZidta AiYdgisj or Oso). Culture solutions suitable for these methods are described in the art, for example, in Catazat ei ai.,..

Arri. VioyesNpo!., 46:117-124, 1979, Uoipo ei ai!., Vioitesnpoi I eiy., 14:863-868, 1992, Apirata apa

Vamipaga, Vgagiyap AgsPpime5 og Vioiodu apa Vioyesppoi., 44:79-88, 2001, US Patents MeMo 3627095, 4379844, 4447530, 4401680, 4526721, 5047332 and 4938972. In some embodiments, any or all of these commercially available culture solutions are published. can be supplemented with biomass substrates obtained as described in this document.

However, in some embodiments, the use of commercially available media is not the most economically feasible option. In such cases, culture solutions can be obtained manually. In some embodiments, culture solutions may contain, in addition to biomass substrates obtained as described in this document, per liter of water at pH 4-7.5: 1.88-2.357 g (MNa)25O, 0.75-1 .5 g of KnegRO", 0.25-5 g of Ma5O4:7HgO, 0.25-0.5 g (Reb5)4:7 HO, 0.25-0.5 27p5O4:7H2O, 0.1-14 ml of solution trace elements. In some embodiments, the culture solution may additionally include 114 mg of boric acid, 480 mg of ammonium molybdate, 780 mg of copper sulfate, and 144 mg of manganese chloride. In some embodiments, the culture solution may additionally contain 0.5 g of yeast extract and may be used to cultivate yeast. In some embodiments, the culture solution can additionally contain 1.0 g of yeast extract and can be used for

From the cultivation of 2 utotopaz tobiiy5. In some embodiments, the culture solution may be adapted for ethanol fermentation and may contain, in addition to the biomass substrates obtained as described herein, per liter of water: sugar equivalent to 80-160 g of glucose, 1 g of KNegRoOx, 1.5 g of MNASI, 0.16 g of MaZO4:7 H2O, 0.08 g of Sasi" and 1.0 g of yeast extract.

In some embodiments, the selected microorganism may be a yeast, and the growth medium may contain, in addition to the biomass substrates obtained as described herein, 1.7 g/L yeast nitrogen base, 2.27 g/L urea, 6.56 g/l of peptone at pH 5.0.

In some embodiments, selected microorganisms can be cultured in the presence of a nitrogen source and/or an additional nitrogen source (for example, when the desired products are proteins or amino acids). In such cases, the nitrogen source may include any nitrogen source, such as livestock waste (eg, bird droppings), human waste, inorganic nitrogen sources, nitrite, nitrate, anhydrous ammonia, ammonium nitrate, diammonium phosphate, monoammonium phosphate, beef, or yeast extract. . In some embodiments, livestock waste and human waste can be sterilized (eg, filtered or autoclaved) prior to use.

Selected microorganisms can be cultivated on a small scale (for example, using standard laboratory equipment and methods known in the field) or on a large scale (for example, using fermentation methods or industrial fermentation). The choice of culture solution depends on the desired scale of cultivation.

Cultivation conditions

Cell culture conditions (eg, temperature, pH, and oxygen requirements) for most organisms are known in the art and can easily be optimized, if desired.

For example, cultivation can be carried out in batch or continuous conditions.

The temperature used for cell cultivation can be chosen according to the selected microorganisms, so as to obtain acceptable yields and conversion rates of the substrate, in particular carbon. Illustrative temperatures are in the range of 25-40 "C. Similarly, the pH value used for cell culture can be maintained in the range at which the selected microorganisms show maximum growth. Illustrative pH ranges are pH 5.0-8.0, such as pH 6, 0-7.0 Additionally, oxygenation levels can be adjusted to maintain 60 a level that provides optimal growth for the selected microorganism. For example, aerobic organisms can be cultured in an oxygenated medium. Alternatively, anaerobic organisms can be cultured in an anaerobic medium.

Methods of cultivation

In some embodiments, selected microorganisms can be cultured without the use of fermentation equipment. For example, a first lignocellulosic biomass material with a first level of recalcitrance can be processed to produce a second material with an altered (eg, reduced) level of recalcitrance. This second material can then be used in the bioconversion stage to obtain a product not present in the first lignocellulosic biomass material. In some embodiments, this second material can be combined (eg, in a liquid medium or in culture) in a cell culture vial with one or more microorganisms under conditions suitable for the growth of microorganisms and product formation. The culture can then be incubated for a period of time sufficient for product formation.

In some embodiments, all cell culture equipment is sterilized or sterile prior to use.

Small-scale methods

In some embodiments, selected microorganisms can be cultured using bench-top fermentation equipment. For example, a first lignocellulosic biomass material with a first level of refractoriness can be processed to produce a second material with an altered (eg, reduced) level of refractoriness. This second material can then be used in the bioconversion stage to obtain a product not present in the first lignocellulosic biomass material. In some embodiments, the second material can be combined with selected microorganisms and cultured in a bench-top fermenter, Vgacp fermenter (V. Vgacp Vioyesii, Auiezrigu, Wysk5) Vioviai IVF with a working volume of 2.8 liters, in a growth medium and under cultivation conditions, suitable for the growth of microorganisms and product formation. The process can then be maintained for a period of time sufficient for product formation. Illustrative parameters set include: temperature 20-45 C; pH 3-9 (which can be maintained by autoclaving); with specific shaking rates and air flow rates (e.g., approximately 1000 rpm and 2 L/min, respectively). In addition, foaming can optionally be suppressed by adding a defoamer at a certain time, for example, an antifoam oil based on polypropylene glycol.

Large-scale methods

In some embodiments, selected microorganisms can be cultured using large-scale fermentation equipment. For example, a first lignocellulosic biomass material with a first level of refractoriness can be processed to produce a second material with an altered (eg, reduced) level of refractoriness. This second material can then be used in the bioconversion stage to obtain a product not present in the first lignocellulosic biomass material. In some embodiments, the second material can be combined with selected microorganisms and cultivated, for example, in a stirred tank bioreactor (eg, in a 300 L stirred tank bioreactor).

Alternatively or additionally, the second material can be combined with selected microorganisms and cultivated in an airlift (pressure cycle) bioreactor (for example, in a 40,000 L airlift bioreactor manufactured by ENM and ISI for OpogpF products)). In both cases, the second material can be combined with selected microorganisms in a culture solution under cultivation conditions suitable for the growth of microorganisms and product formation. The process can then be maintained for a period of time sufficient for product formation.

In some embodiments, the selected microorganisms can be cultured using fed-batch fermentation (eg, fixed-volume fed-batch fermentation or variable-volume fed-batch fermentation) in which nutrients are added in a controlled manner according to the needs of the culture solution (see Fig. 44 and Fig. 45). In the process of serial fermentation with feeding, growth-limiting substrates are added to the culture solution in a highly concentrated form or in a gaseous form that does not change the volume of the culture solution. After the fermentation reaches a certain stage, the volume of the culture solution can optionally be removed and replaced with a fresh culture solution. At this stage, the volume of culture solution not removed from the fermenter serves as the starting culture for the next cycle, and the removed volume contains the desired product. This process is known in the industry as cyclic cultivation with feeding for a culture with a fixed volume. One of the advantages of using fed-batch for a fixed-volume culture is that the desired products can be obtained before the fermentation process is complete. In addition, cyclic cultivation with feeding for the cultivation process with a fixed volume can be continuous. In the process of batch fermentation with feeding with a variable volume, growth-limiting substrates are added as necessary to stimulate further growth of the culture at a concentration equal to that of the original culture. Therefore, the total volume of culture is growing. This process can be repeated until the culture volume reaches the capacity of the fermenter. In this method, larger containers for fermentation are preferable, since such containers contain larger volumes of culture solution. Then the desired products can be obtained from the culture solution, for example, at the end of the fermentation process. Both of these fed-batch processes allow for optimal yields and productivity. In some embodiments, the process may include providing continuously oxygenated water, for example, using an airlift fermentation system.

Batch processes with feeding are also described in European patent application Mo 533039.

After fermentation, the selected microorganism and/or product can be collected and optionally isolated and/or purified. Methods of collecting microorganisms from culture solutions include, for example, centrifugation and/or filtration.

Additional processing to obtain food products

Cultures for use, for example, as ingestible food for animals and/or humans can be further processed, for example, using the methods described in US patents MoMo 5935841; 6270816; 5980958 and 3809614. Alternatively or additionally, the collected organism can be processed to reduce the content of nucleic acids in it, for example, using the method of the British patent Mo 1440642; separate, if desired, for example, using the method of British patent Mo 1473654, or by filtration or centrifugation; and it is possible to modify its taste, for example, using the methods of British patents MoMo 1508635; 1502455; 1496113 and/or 1587828.

Humans lack the enzyme uricase to catalyze the conversion of uric acid to soluble allantoin. Consumption of microbial cells that contain high levels of nucleic acid can therefore lead to elevated uric acid levels and associated complications in humans. Thus, in some embodiments, nucleic acids can be removed or can be reduced in samples containing microbial cells or food products derived from microbial cells (e.g., proteins, fats, oils, and carbohydrates) prior to human consumption, e.g., with using the methods described in I amloga and I em/i5 (US patent Mo 4330464). In some embodiments, nucleic acids can be removed or can be reduced in samples containing microbial cells or food products derived from microbial cells (e.g., proteins, fats, oils, and carbohydrates) prior to human consumption, e.g., using methods described in US patent No. 6,270,816. For example, microbial cells can be destroyed and simultaneously reduce the amount of their nucleic acids by rapidly heating the culture solution to at least 60 "C. This process can be used to stimulate the loss of viability and the release of some of the cellular nucleic acids (for example, DNA and RNA) into the supernatant After heating, the culture solution can be centrifuged and washed to remove nucleic acids.

In some embodiments, the ratio of protein:"RNA for protein in the human consumption sample should be at least 12:1. In some embodiments, the total nucleic acid content of the human consumption sample can be reduced to about 2 95 (eg, 2 9o, less than 2 90, 0.1-2.0 90, 0.1-1.5 V, 0.1-1 90, 0.1-0.5 95, 0.1-0.3 90, 0.1 Fo ) of the dry weight of the sample.

In some embodiments, nutritional evaluation and/or toxicological evaluation of samples containing microbial cells or food products derived from microbial cells (e.g., proteins, fats, oils, and carbohydrates) may be performed prior to consumption by animals (e.g., for each specific species). .

In some embodiments, the microbial proteins can be dried, lyophilized, or they can be in solution, and they can be present in isolated form or in the presence of one or more additional food sources.

In some embodiments, samples containing microbial cells or food products derived from microbial cells (eg, proteins, fats, oils, and carbohydrates) can be made into nutritional gels. The quality of the gel can be evaluated using stress and tension tests, for example, using the torsion method of UMy ei ai., U. Tech. Zitsaiyev, 16:53-74 (1985) or with the help of a Kpeo Tech AR-83 helemeter (Bip bsiepsez So. Zeashe, MA, OBA).

As a rule, stress values (elastic component) greater than 1.9 to 2.0 and tensile values of 30-35 kPa are a reliable indication of gel strength. for In some embodiments, samples containing microbial cells or food products

derived from microbial cells (e.g., proteins, fats, oils, and carbohydrates) can be flavored and/or colored, for example, to increase the palatability of these species.

In some embodiments, samples containing microbial cells or food products derived from microbial cells (eg, proteins, fats, oils, and carbohydrates) can be used as meat analogs or in the preparation of meat analogs. "Meat analog" is an industry term for meat substitutes or synthetic meats made primarily from a non-animal source, such as plant proteins.

In some embodiments, the health and nutritional value of foods derived from microbial cells (eg, proteins, fats, oils, and carbohydrates) described herein are considered prior to consumption by animals and/or humans.

Composition of products

In some embodiments, the food products described herein can be used as food products or for their preparation (eg, solid or liquid food products). In some embodiments, the food products can be used individually or can be combined. In some embodiments, food products can be combined with materials that improve texture (eg, wheat protein). In some embodiments, the food products described herein can be manufactured as meat alternatives (see, e.g., OyogpF manufactured by Magi/Bosa5, OK). In some embodiments, the food products described herein can be combined with other proteins, protein sources, or foods, such as mycoprotein, textured vegetable protein, tofu, tempeh, miso, soy products, and/or wheat protein.

In some embodiments, any of the products and co-products described herein can be combined with flavors and/or colorants, such as fine chemical flavors and aromas.

Water treatment

In the processes described in this document, when water is used in any process, it may be domestic waste water, such as municipal waste water, or fecal water. In some embodiments, domestic or fecal water is sterilized prior to use.

3. Sterilization may be performed using any desired method, such as irradiation, steam, or chemical sterilization.

EXAMPLES

The examples presented below are intended to illustrate and not limit the ideas of this description.

Example 1 - Production of fibrous material from paper with a multilayer coating

A 1,500 lb (680 kg) slipway of clear half-gallon (1.9 L) juice cartons made of unprinted white kraft paperboard having a bulk density of 20 lb/ft3 (0.32 g/cm3), obtained from Ipiegpaiop! Rare Each carton is folded into a flat state and then fed into a Z Priipsp Vatsoy shredder at a rate of approximately 15 to 20 pounds per hour (6.8-9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 in. (2.5 cm) and a thickness equivalent to that of the parent material (approximately 0.075 in. (0.2 cm)).

The confetti-like material is fed into a Mipsop Model 5230 Rotary Knife Cutter. The Model 5230 is equipped with four rotating blades, four fixed blades, and a discharge screen that has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The fibrous material has a VET surface area of 0.9748 mg/g w/- 0.0167 me/g, a porosity of 89.0437 905 and a bulk density (at 0.53 psi abs. (3.7 kPa) ) 0.1260 g/ml. The average fiber length is 1.141 mm and the average fiber width is 0.027 mm, giving an average G/O of 42:1. A picture obtained with the help of a scanning electron microscope at a magnification of 25X of the fibrous material is presented in Fig. 26.

Example 2 - Production of fibrous material from bleached kraft cardboard

A slipway weighing 1500 lb (680 kg) of pure bleached kraft paperboard having a bulk density of 30 lb/ft? (0.48 g/smu), obtained from Ipiegpaiopa! Rare The material is folded into a flat state and then fed into the shredder at a rate of approximately 15 to 20 pounds per hour (6.8-9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 in. (2.5 cm) and a thickness equivalent to that of the parent material (approximately 0.075 in. (0.2 cm)). The candy-like material is fed into a Mipsop model 5230 rotary knife cutter. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between and fixed rotating blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The fibrous material has a VET surface area of 1.1316 mg/g w/- 0.0103 mg/g, a porosity of 88.3285 95 and a bulk density (at 0.53 lb/sq. in. abs. (3.7 kPa) ) 0.1497 g/ml. The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average G/O of 43:1. A picture obtained with the help of a scanning electron microscope at a magnification of 25X of the fibrous material is presented in Fig. 27.

Example C - Production of twice-shredded fibrous material from bleached kraft cardboard

A slipway weighing 1500 lb (680 kg) of pure bleached kraft paperboard having a bulk density of 30 lb/ft? (0.48 g/smu), obtained from Ipiegpaiopa! Rare The material is flattened and then fed into a Z Pr Ripsp Watersop shredder at a rate of approximately 15 to 20 pounds per hour (6.8-9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The grinding material coming out of the device looks like confetti (see above). The candy-like material is fed into a Mipsop rotary knife cutter, Model 5230. The discharge screen has 1/16 inch (0.16 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A cutting device with a rotating knife crushes confetti-like fragments with points

From the blade, breaking the fragments and releasing the fibrous material at a rate of about one pound per hour (454 grams per hour). The material obtained after the first crushing is again fed into the same plant described above and crushed again. The resulting fibrous material has a VET surface area of 1.4408 mg/g w/- 0.0156 mg/g, a porosity of 90.8998595 and a bulk density (at 0.53 lb/sq. in. abs. (3.7 kPa) ) 0.1298 g/ml. The average fiber length is 0.891 mm and the average fiber width is 0.026 mm, giving an average G/O of 34:1. A picture obtained with the help of a scanning electron microscope at a magnification of 25X of the fibrous material is presented in Fig. 28.

Example 4 - Production of three-fold shredded fibrous material from bleached kraft cardboard

A slipway weighing 1500 lb (680 kg) of pure bleached kraft paperboard having a bulk density of 30 lb/ft? (0.48 g/cm3), obtained from Ipiegpaiopa! Rare The material is flattened and then fed into the Z Pr Ripsp Vacoi shredder at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The material coming out of the shredder resembled confetti (see above). The candy-like material is fed into a Mipsop model 5230 rotary knife cutter. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A cutting device with a rotating knife crushes confetti-like fragments with the edges of the blades. The material obtained after the first crushing is again fed into the same plant described above, and the sieve is replaced with a sieve with holes of 1/16 inch (0.16 cm). The material is crushed.

The material obtained after the second crushing is again fed into the same plant described above, and the screen is replaced with a screen with openings of 1/32 inch (0.08 cm). This material is crushed. The resulting fibrous material has a VET surface area of 1.6897 mg/g /- 0.0155 me/g, a porosity of 87.7163 95 and a bulk density (at 0.53 psi abs. (3.7 kPa) ) 0.1448 g/ml. The average fiber length is 0.824 mm and the average fiber width is 0.0262 mm, giving an average G/O of 3271.

A picture obtained with the help of a scanning electron microscope at a magnification of 25X of the fibrous material is presented in Fig. 29. bo Example 5 - Obtaining compacted fibrous material from bleached kraft paper without adding a binder

The fibrous material is prepared according to Example 2. Approximately 1 pound (454 g) of water is sprayed for every 10 pounds (4540 g) of fibrous material. The fibrous material is compacted using a Saigoppia Reimei MiiII 1100 pellet press operating at 75"b. Pellets having a bulk density in the range of about 7 lb/ft (0.11 g/cm3) to about 15 lb/ft are obtained. (0.24 g/cm3).

Example 6 - Production of compacted fibrous material from bleached kraft cardboard with a binder

Fibrous material is obtained according to example 2.

Prepare a stock solution of ROI MOH "M UUZEA M10 (polyoxyethylene) in water at a concentration of 2 wt. 95.

Approximately 1 pound (454 g) of stock solution is sprayed for every 10 pounds (4540 g) of fibrous material. The fibrous material is compacted using a granulation press

1100, operating at 75°C. Granules having a bulk density in the range of about 15 lb/ft" (0.24 g/cm) to about 40 lb/ft" (0.64 g/cm cm3).

Example 7 - Reduction of the molecular mass of cellulose in fibrous kraft paper by gamma radiation with minimal oxidation

The fibrous material is prepared according to example 4 and then compacted according to example 5.

Compacted granules are placed in a glass ampoule with a maximum capacity of 250 ml. The glass ampoule is evacuated under a high vacuum (105 torr) for 30 minutes, and then backfilled with argon gas. The ampoule is sealed under argon. The granules in the ampoule are irradiated with gamma radiation for about 3 hours at a dose level of about 1

Mrad per hour with the production of irradiated material in which the cellulose has a lower molecular weight than in the source material of fibrous kraft paper.

Example 8 - Reduction of the molecular mass of cellulose in fibrous kraft paper by gamma radiation at maximum oxidation

The fibrous material is prepared according to example 4 and then compacted according to example 5.

Z Compacted granules are placed in a glass ampoule with a maximum capacity of 250 ml. The glass ampoule is sealed in an air atmosphere. Granules in the ampoule are irradiated with gamma radiation for about 3 hours at a dose level of about 1 Mrad per hour to produce irradiated material in which the cellulose has a lower molecular weight than in the source material of fibrous kraft paper.

Example 9 - Methods of determining the molecular weight of cellulosic and lignocellulosic materials using gel permeation chromatography

Cellulosic and lignocellulosic materials for analysis are processed according to example 4.

Sample materials presented in the following tables include kraft paper (P), wheat straw (M/5), alfalfa (A), and millet (503). The number "132" in the sample ID refers to the particle size of the material after passing through a 1/32 inch (0.08 cm) sieve. The number after the hyphen refers to the dose of radiation (Mrad) and "05" refers to ultrasonic treatment. For example, the IU sample "P132-10" refers to kraft paper, which was subjected to crushing to the size of particles of caliber 132 and irradiated with a dose of 10 Mrad.

Table 1

Peak average molecular weight of irradiated kraft paper deviation

Kraft paper |RIZ2 | 77/70 777777 no | 77 Z2853a10006 79 110000 DFRYZAMO | 70770111 71111111 bi39vzho4681shШ 11111000 FRIZOLOOYI | 70004171 8444а2580.4...:К КК: dust SSZ IN ETO PO EX PNYA BOTH POLON 2 15 Ei I 11111111 DFRIZ2OV | 07777171 yes///// | 777777 zoo5mO3 "k Low doses of radiation appear to increase the molecular weight of some materials

Dosing level - 1 Mrad/h hTreatment for 30 minutes with 20 kHz ultrasound using a 1000 W horn in recirculation conditions where the material is dispersed in water.

Table 2

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IBa132-100-05| 1 | forehead HER 77777777171711717111111111111469641465...:.:SU "Peaks merge after processing ""Low doses of radiation appear to increase the molecular weight of some materials

Dosage level - 1 Mrad/hour 2 Treatment for 30 minutes with 20 kHz ultrasound using a 1000 W horn in recirculation conditions where the material is dispersed in water.

Gel permeation chromatography (GPC) is used to determine the molecular weight distribution of polymers. During CPS analysis, a polymer sample solution is passed through a column filled with a porous gel that traps small molecules. The sample is separated based on molecular size, with larger molecules eluting faster than smaller molecules. The retention time of each component is most often determined by refractive index (RI), evaporative light scattering (EI5) or ultraviolet radiation (M) and compared with a calibration curve. The resulting data are then used to calculate the molecular weight distribution for the sample.

Molecular weight distribution is used to characterize synthetic polymers instead of individual molecular weight. Statistical averages are used to characterize this distribution. The most common of these average values are "number average molecular weight" (My) and "weight average molecular weight" (Mu). Methods of calculating these values are described, for example, in example 9 PCT/O5/2007/022719.

The polydispersity index or RI is defined as the Mu/My ratio. The higher the RI, the wider or more dispersed the distribution. The lowest value that RI can have is 1. It corresponds to a monodisperse sample, that is, a polymer in which all molecules in the distribution have the same molecular weight.

The peak value of the molecular weight (Mp) is another descriptive feature defined as the mode of the molecular weight distribution. It means the molecular weight that is most common in the distribution. This value also gives an idea of the molecular weight distribution.

Most CPS measurements are performed against various polymer standards. The accuracy of the results depends on how close the characteristics of the analyzed polymer are

Match the characteristics of the used standard. The expected reproducibility error between different series of individually calibrated determinations is approximately 5-10 95 and is typical of the limited precision of CPS determinations. So,

ODS results are most useful when comparisons are made between molecular weight distributions of different samples in the course of one series of determinations.

Lignocellulosic samples require preparation before SPC analysis. First, prepare a saturated solution (8.4 95 by mass) of lithium chloride (LIS!) in dimethylacetamide (OMAs). About 100 mg of each sample is added to about 10 g of freshly prepared saturated solution of HICI/OMAsC, and the mixtures are heated to about 150-170 °C with stirring for 1 hour. The resulting solutions are mainly light yellow to dark yellow in color. The temperature of the solutions is reduced to approximately 100 "C and they are heated for an additional 2 hours. The temperature of the solutions is then reduced to about 50 °C and the sample solutions are heated for about 48 to 60 hours. It should be noted that the samples irradiated at 100 Mrad are more easily solubilized compared to their untreated counterparts. In addition, the crushed samples (indicated by the number 132) have a slightly lower average molecular weight compared to uncut samples.

The obtained solutions of the samples are diluted 1:11 using ХОМас as a solvent and filtered through a 0.45-μm RTEE filter. Then the filtered solutions of the samples are analyzed using SPC. The peak average molecular weight (Mw) of the samples, as determined by gel permeation chromatography (GPC), is summarized in Tables 1 and 2, as indicated above, under the analysis conditions presented in Table 3. Each sample is prepared in duplicate, and each preparation the sample is analyzed in two parallels (two injections), with a total of four injections per sample. To obtain a calibration curve for the molecular weight scale from approximately 580 to 750,000 Daltons, ROTA polystyrene standards are used and

RBO1V EazisSai.

Table C

Conditions of ORS analysis

Example 10 - Determination of crystallinity of irradiated material using X-ray diffraction

X-ray diffraction (XD) is a method by which a crystalline sample is irradiated with monoenergetic X-rays. The interaction of the lattice structure of the sample with these X-rays is recorded, and it provides information about the crystalline

From the irradiated structure. The obtained characteristic "print" allows the identification of crystalline compounds present in the sample. Using pattern matching analysis (She Kieimeiyi Keiipetepi) quantitative analyzes of samples containing more than one crystalline compound can be performed.

Each sample is placed on a holder with a zero background and placed in a diffractometer

RUM1800, which uses radiation of Si. Scans are then performed in the range from 57 to 507 with a step size of 0.05" and a counting time of 2 hours in each case.

After obtaining the diffractograms, the phases are identified using the Romaeg Oiitasinop Riie published by Ipiegpaiopai Sepige tog Oiitasiiop BVaia. In all samples, the identified crystalline phase is cellulose - Ia, which has a triclinic structure.

Distinctive features for 20 samples are the peak width, which is related to the size of the crystal domain. The experimental peak width is used to calculate the domain size and percent crystallinity, which are presented in Table 4.

Table 4

KKO data including domain size and 95 crystallinity

Percent crystallinity (Xe 95) is defined as the ratio of the area for the crystalline phase to the total area under the X-ray diffraction peaks, and it is equal to 100905 x (As/Aa--As), where

Ac is the area for the crystalline phase,

Aa is the area for the amorphous phase,

Heh x percentage of crystallinity.

To determine the percent crystallinity of each sample, it is necessary to first determine the amount of the amorphous phase. This is done by estimating the area of each diffractogram, which can be attributed to the crystalline phase (which is shown by the sharper peaks) and the non-crystalline phase (which is shown by the broad humps below the pattern and centered at 227 and 387).

A systematic process is used to minimize error in these calculations due to broad crystal peaks as well as high background intensity. First, a linear background is applied, and then it is canceled. Second, each of the two Gaussian peaks centered at 227 and 387 with a width of 10-12" is aligned with the bumps under the crystalline peaks. Third, the area under the two broad Gaussian peaks and the rest of the pattern is determined. Finally, the percentage is calculated crystallinity by dividing the area under the crystalline peak by the total intensity (after background subtraction).The domain size and 95 crystallinity of the samples as determined by X-ray diffraction (XRD) are presented in Table 4, above.

Example 11 - Porometry analysis

Mercury analysis of pore size and pore volume (Table 5) is based on passing mercury (a non-wetting liquid) into a porous structure under strictly controlled pressure. Since mercury does not wet most substances and does not penetrate the pores spontaneously due to capillary action, it must be driven into the cavities of the sample using external pressure. The pressure required to fill the cavities is inversely proportional to the pore size. Only a small amount of force or pressure is required to fill large cavities, while much higher pressure is required to fill very small pores.

Table 5

Distribution of the size and volume of pores using mercury porometry. Sere- Sere-. Average O'bemna Imaginary

General with dinny dinny: density at 10 penetrating diameter | diameter diameter 0.50 lb./sq. (skeletal) - pore area and density of 1 sample pore pore (me/g) pore pore (4M/A) inch.abs. Porosity volume ml/g (area) | (area) (3.45 kPa) (μm) (g/ml) (μm) (μm) (g/ml)

The ASHOROGE 9520, a device for determining the density of pores, can reach a maximum pressure of 414 MPa or 60,000 psi. inch abs It has four low pressure stations for sample preparation and macropore data collection at from 0.2 psi. inch abs (1.4 kPa) to 50 lb/sq. inch abs (345 kPa). It has two high-pressure chambers that collect data at from 25 psi. inch abs (172 kPa) to 60,000 lb/sq. inch abs (414 MPa). The sample is placed in a cup-shaped device called a penetrometer, which is connected to a glass capillary rod with a metal coating. As mercury penetrates the cavities in and around the sample, it moves down the capillary rod. The loss of mercury from the capillary rod leads to a change in capacitance. The change in electrical capacity during the experiment is converted into the volume of mercury, based on the known volume of the rod of the used penetrometer. A variety of penetrometers are available with different bowl (sample) and capillary sizes to accommodate most sample sizes and configurations. Table 6 below identifies the key parameters calculated for each sample.

Table 6

Definition of parameters

Total penetrating volume | The total volume of mercury penetrating during the experiment. It may include interstitial filling between small particles, sample porosity and sample compression volume assuming a cylindrical pore shape (area)

Bulk density The mass of the sample divided by the total volume. Full volume is defined at a fill pressure of typically 0.5 psi (3.45 kPa)

Apparent Density The mass of the sample divided by the volume of the sample measured at the highest pressure, typically 60,000 psi abs (414

MPa)

Example 12 - Particle size analysis

The method of determining particle size using light scattering is based on theory

Mee (which also covers Fraunhofer's theory). Mie's theory predicts the intensity-angle relationship as a function of the size of the spherical scattering particles, provided that the other system variables are known and held constant. These variables are the wavelength of the incident light and the relative refractive index of the sample material. The application of Mie's theory provides detailed information on particle size. Table 7 summarizes the size of the particles when used as parameters of median diameter, average diameter and modal diameter.

Table 7

Particle size during laser radiation scattering (dry sample dispersion) (μm) (μm) (μm)

Particle size is determined by laser scattering (dry sample dispersion) using a MaiYimet Mawiegwileg 2000, using the following conditions: feed rate: 35,905, disperser pressure: 4 bar, optical model: (2.610, 1.000), 1.000.

The appropriate amount of the sample is fed to the vibrating tray. The feed rate and air pressure are adjusted to ensure that the particles are properly dispersed.

The key component is choosing an air pressure that breaks up the agglomerates but does not compromise the integrity of the sample. The amount of sample required varies with particle size. As a general rule, samples with fine particles require less material than samples with large particles.

Example 13 - Surface area analysis

The surface area of each sample is analyzed using the Misgotegys5 AZAR 2420 AssayIegagei Agea apa Rogoziteigu Zubiet system. The samples are prepared by primary degassing for 16 hours at 40 "C. Next, the free space (both warm and cold) with helium is calculated, and then the tube with the sample is again evacuated to remove the helium.

Data collection begins after this second vacuum and consists of establishing a set pressure that controls how much gas is dosed into the sample. At each given pressure, the amount of adsorbed gas and the true pressure are determined and recorded. The pressure inside the sample tube is measured using a pressure sensor. Additional doses of gas continue to arrive until the set pressure is reached and equalization is ensured. The amount of adsorbed gas is determined by summing the set of doses for the sample. Pressure and quantity determine the gas adsorption isotherm and are used to calculate a number of parameters, including surface area

VET (table 8).

Table 8

Generalized representation of surface area by gas adsorption

The BET model for isotherms is a widely used theory for calculating the specific surface area. The analysis involves determining the monolayer capacity of the sample surface by calculating the amount needed to cover the entire surface with one close-packed layer of krypton. To determine the total surface area, the capacity of the monolayer is multiplied by the cross-sectional area of the sample gas molecule. The specific surface area is the surface area of a sample aliquot divided by the mass of the sample.

Example 14 - Determination of fiber length

Fiber length distribution testing was performed in three parallels on the presented samples using the Tesprar Mogri GVO1 system. The average length and width are presented in Table 9.

Table 9

Generalized presentation of data on the length and width of lignocellulosic fibers

Arithmetic average length, | Statistically adjusted Width

IO sample average value | weighted along the length | average length weighted (mm) (mm) (mm) lengthwise (mm)

Example 15 - Ultrasonic treatment of irradiated and non-irradiated millet

Millet is crushed according to example 4. Millet is treated only with ultrasound or irradiation with gamma rays at a dose of 10 or 100 Mrad followed by ultrasound treatment.

The resulting materials correspond to 5132-VC (unirradiated), 5132-10-VC (10 Mrad and sonicated), and 5132-100-VC (100 Mrad and sonicated), as shown in Table 1.

Ultrasound treatment is carried out for each sample for 30 minutes using ultrasound with a frequency of 20 kHz from a 1000 W horn in recirculation conditions. Each sample is dispersed in water at a concentration of approximately 0.10 g/ml.

In Fig. 30 and 31 present the device used for ultrasonic treatment.

The device 500 includes a transducer 502 connected to a booster 504 connected to a horn 506 made of titanium or a titanium alloy. The horn, which has an insulation 510 made of MITOMF around its perimeter on its processing side, forms a liquid-tight seal with the processing cell 508. The processing side of the horn is immersed in a liquid, such as water, in which the sample to be processed is dispersed. ultrasound treatment. The pressure in the cell is monitored using the pressure sensor 512. In the working state, each sample is pushed by the pump 517 from the container 516 through the processing cell and treated with ultrasound. After sonication, the sample is collected in container 520. The process may be reversible in that the contents of container 520 may be sent through the processing cell and collected in container 516. This process may be repeated several times until the desired level of sample processing is achieved.

Example 16 - Scanning electron microscope images of non-irradiated millet compared to irradiated and irradiated and sonicated millet

Millet samples for scanning electron microscope images are placed on carbon tape and sputter coated with gold (70 seconds). Images are obtained using a scanning electron microscope with field emission of UEOI. 6500.

In Fig. 32 presents a picture obtained with the help of scanning electronic

From a microscope at 1000X magnification, fibrous material obtained by crushing millet on a rotary knife cutter and then passing the crushed material through a 1/32 inch (0.08 cm) sieve.

In Fig. 33 and 34 show images obtained using a scanning electron microscope at a magnification of 1000X of the fibrous material from Fig. 32 after irradiation with gamma rays at a dose of 10 and 100 Mrad, respectively.

In Fig. 35 presents images obtained using a scanning electron microscope at a magnification of 1000X of the fibrous material from Fig. 32 after 10 Mrad irradiation and ultrasound treatment.

In Fig. 36 presents images obtained using a scanning electron microscope at a magnification of 1000X of the fibrous material from Fig. 32 after 100 Mrad irradiation and ultrasound treatment.

Example 17 - Infrared spectrum of irradiated kraft paper compared to non-irradiated kraft paper

ET-IR analysis was performed using standard methods on a Misoietras 400.

The results indicate that all samples presented in Table 1 correspond to cellulose-based material.

In Fig. 37 shows the infrared spectrum of kraft paper shredded according to example 4, and Fig. 38 presents the infrared spectrum of the kraft paper from Fig. 37 after exposure to gamma radiation at a dose of 100 Mrad. The irradiated sample exhibits an additional peak in region A (centered at approximately 1730 cm") that is not present in the unirradiated material.

Example 18 - Combination of pretreatment with an electron beam and ultrasound

Millet is used as raw material and crushed with a cutting device with a rotating knife

Mipsop on fibrous material. The fibrous material is then spread evenly on an open tray made of tin with an area of more than 500 in: (12.7 me). The fibrous material is distributed to a thickness of approximately 1-2 inches (2.5-5 cm) in an open tray. The fibrous material can be distributed in plastic bags at lower radiation doses (less than 10 Mrad) and left exposed on a metal tray at higher radiation doses.

Then individual samples of fibrous material are subjected to successive doses of electron beam irradiation to achieve a total dose of 1, 2, 3, 5, 10, 50 and 100 Mrad. Some samples are maintained under the same conditions as other samples but not irradiated to serve as controls. After cooling, the irradiated fibrous material is sent for further processing through an ultrasonic treatment device.

The ultrasonic treatment device includes a transducer connected to a booster connecting a horn made of titanium or a titanium alloy. The horn, which has insulation made of MITOMO' around its perimeter on its processing side, forms a liquid-tight seal with the processing cell. The processing side of the horn is immersed in a liquid, such as water, in which the sample to be sonicated is dispersed.

The pressure in the cell is monitored using a pressure sensor. In operation, each sample is pumped from the tank through the processing cell and sonicated.

For the preparation of irradiated fibrous material for sonication,

The irradiated fibrous material is extracted from any container (for example, plastic bags) and dispersed in water at a concentration of approximately 0.10 g/ml. Ultrasound treatment is carried out on each sample for 30 minutes using ultrasound with a frequency of 20 kHz from a 1000 W horn in recirculation conditions. After ultrasonic treatment, the irradiated fibrous material is collected in a container. This process can be repeated many times until the desired level of processing is achieved, based on the monitoring of structural changes in the millet. Also, some irradiated samples are kept under the same conditions as other samples but not sonicated to serve as controls. In addition, some samples that were not irradiated or not treated with ultrasound also act as controls. Thus, some controls are not treated, some are only irradiated, and some are only treated with ultrasound.

Example 19 - Testing of pre-treated biomass using microorganisms

Specific lignocellulosic materials, pretreated as described herein, are analyzed for toxicity to common strains of yeast and bacteria used in the biofuel industry for the fermentation step of producing ethanol. In addition, sugar content and compatibility with cellulase enzymes are investigated to determine the effectiveness of the processing process. Testing of pre-treated materials is carried out in two phases as follows.

I. Toxicity and sugar content

The toxicity of pre-treated herbs and paper raw materials is determined in yeast

Zasspagotusev segemisiae (wine yeast) and Rispia 5iiirie (ATSS 66278), as well as in bacteria 2utotope5z torbiii5 (ATSS 31821) and Siovigidiit "(peptosePysht (ATSS 31924). For each of the organisms, growth studies are conducted to determine the optimal incubation time and sample collection.

Then each raw material is incubated, in two parallels, with 5. segemiviae, R. 5iiryiv, 7. torbiiyiv and S.

Shegtoseiisht, in a standard microbiological environment for each organism. For two yeast strains, Z. segemibiae and R. Ziiryich, UM broth is used. For 7. tobiii5 use KM medium and for S. IpeptosePysht use CMA4 medium. For comparison, a positive control is used, with the addition of pure sugar, but without raw materials. In the incubation process, during the 12-hour period, a total of five samples are obtained at 60 time points of 0, 3, 6, 9, and 12 hours and analyzed for viability (counting in cups for 2. thoria and direct counting for 5. segemiviae) and ethanol concentration.

The sugar content in raw materials is determined using high-performance liquid chromatography (HPLC), using a Zpodeh?F zidag 5РО810 column, or a Viogai column

Atipeh? NRH-87R. Each raw material (approximately 5 g) is mixed with reverse osmosis (RO) water for 1 hour. The liquid part of the mixture is removed and analyzed for the content of glucose, galactose, xylose, mannose, arabinose and cellobiose. The analysis is carried out according to the protocol

Oeieptipaiyop oi 5igisiiga! Sagbopuagaieez apa Gidpip ip Wiotav5 of Mayopai! Vioepegdu Sepieg.

II. Cellulase compatibility

The raw material is tested, in two parallels, with the commercially available enzyme complex AssayIegazef 1000, which contains an enzyme complex that reduces lignocellulosic biomass to fermentable sugars, including two different cellulase preparations, Thispodegta geeseii and Avregoii5 podshiap5, at the recommended temperature and concentration in the vial

Epepteueg. Vials are incubated with moderate shaking at approximately 200 rpm. within 12 hours. During this period of time, samples are taken every three hours at time points 0, 3, 6, 9 and 12 hours to determine the concentration of reducing sugars (Nore apa Oeap, Vioiesp -., 1974, 144:403) in the liquid part of the vials.

Example 20 - Production of alcohol using pretreatment in the form of irradiation - ultrasound treatment

The optimal size of biomass conversion plants is influenced by factors including economies of scale and the type and availability of biomass used as feedstock.

Increasing plant size tends to increase the economies of scale associated with manufacturing processes. However, increasing plant size also tends to increase costs (e.g., transportation costs) per unit of biomass feedstock. Studies analyzing these factors indicate that an acceptable biomass conversion plant size can be in the range of 2,000 to 10,000 dry tons of biomass feedstock per day. The installation described below is scaled to process 2,000 tons of dry biomass feedstock per day.

In Fig. 39 presents a technological diagram of a biomass conversion system adapted for millet processing. The raw material preparation subsystem processes raw materials

From biomass to remove foreign objects and provides particles of constant size for further processing. The pretreatment subsystem changes the molecular structure (eg, reduces the average molecular weight and crystallinity) of the biomass feedstock by irradiating the biomass feedstock, mixing the irradiated biomass feedstock with water to form a slurry, and applying ultrasonic energy to the slurry. Irradiation and ultrasound treatment transform cellulosic and lignocellulosic components of biomass raw materials into fermentable materials. The primary processing subsystem ferments glucose and other low molecular weight sugars present after pretreatment to form alcohols.

Preparation of source material

The selected level of the given raw material for the enterprise is 2000 tons of dry millet biomass material per day. The specified starting material is chopped and/or crushed millet.

Biomass raw material, in the form of bundles of millet, is delivered to the installation. In some cases, the millet bundles are wrapped in plastic netting to ensure that they do not fall apart during transport, and they may also be wrapped in plastic film to protect the bundle from the weather. Ties have either a square or round shape. Bundles are delivered to installations from remote storage on large trucks with trailers. When trucks arrive, they are weighed and unloaded with forklifts.

Some bundles are sent to local storage, while others are placed directly on conveyors.

Since millet is only available seasonally, long-term storage is required to supply the plant with raw material year-round. Long-term storage may consist of 400-500 acres of uncovered clustered bonds in an area (or areas) reasonably close to the ethanol production facility. Local short-term storage, equivalent to 72 hours of production, is provided in open areas. Ties and surrounding driveways, as well as transport conveyors are on a concrete slab. The concrete slab is used due to the necessary volume of freight traffic for the delivery of the necessary large amount of biomass raw materials. The concrete slab minimizes the amount of standing water in the storage area, as well as reducing the impact on the raw biomass of dirt. Stored material provides short-term supply for weekends, holidays and when normal direct delivery of material for processing is interrupted. because the bundles are unloaded by forklifts and placed directly on the conveyors for transporting the bundles or on the short-term storage area. Bundles are also removed from short-term storage by forklifts and placed on conveyors for bundle transportation.

The bundles are sent to one of two stations for deployment. The unfolded bundles are broken using a straightening bar and then unloaded onto a conveyor that passes a magnetic separator to remove metal before crushing. A doped iron magnet is provided to capture stray magnetic metal, and a receiving screen removes oversize and foreign material before a plurality of slicer-shredder systems reduce the size of the biomass feedstock to the proper size for pretreatment. Slicing device-crushing device systems include slicing devices and rotary knife cutting devices. Slicing devices reduce the size of the raw biomass raw material and feed the resulting material into cutting devices with a rotating knife. Cutting devices with a rotating knife simultaneously crush raw biomass and sift the resulting material. At the end, the raw biomass can be transported to the system for pre-treatment.

To limit total downtime due to required maintenance and/or system interruptions, three storage silos are provided for raw material preparation. Each silo can hold approximately 55,000 cubic feet (1,560 mu) of biomass feedstock (43 plant hours).

Preliminary processing

The conveyor belt moves the biomass feedstock from the raw material preparation subsystem 110 to the pretreatment subsystem 114. As shown in FIG. 40, in the pre-treatment subsystem 114, biomass raw materials are irradiated using electron beam emitters, mixed with water to form a suspension and exposed to ultrasonic energy. As discussed above, irradiation of biomass raw materials changes the molecular structure (for example, reduces refractoriness, average molecular weight, and crystallinity) of biomass raw materials. Mixing the irradiated biomass raw material into a suspension and applying ultrasonic energy to the suspension further changes the molecular structure of the biomass raw material.

The use of sequential radiation exposure and ultrasound treatment can have

Synergistic effects, since the combination of methods appears to provide more significant changes in molecular structure (eg, reduced recalcitrance, average molecular weight, and crystallinity) than changes that can be effectively provided by either method alone. Without regard to theory, in addition to reducing the polymerization of the biomass feedstock by breaking the intramolecular bonds between the segments of the cellulosic and lignocellulosic components of the biomass feedstock, irradiation can make the overall physical structure of the biomass feedstock more fragile. After mixing the brittle biomass feedstock into a suspension, the application of ultrasonic energy further changes the molecular structure (for example, lowers the average molecular weight and crystallinity) and can also reduce the particle size of the biomass feedstock.

Irradiation with a beam of electrons

A conveyor belt 491 carrying the biomass feedstock to the pretreatment subsystem distributes the biomass feedstock into a plurality of feedstock streams (eg, 50 feedstock streams), each of which leads to individual electron beam emitters 492. In this embodiment, the biomass feedstock is irradiated in dry state Each stream of source material is transferred on a separate conveyor belt to its associated electron beam emitter. Each conveyor belt for irradiation of raw materials can be approximately one meter wide. Before reaching the electron beam emitter, localized vibration is induced in each conveyor belt for uniform distribution of dry biomass raw material across the transverse width of the conveyor belt.

The electron beam emitter 492 (eg, electron beam irradiators commercially available from Tyap Sogrogayop, Zap Oiedo, CA) is adapted to use an electron dose of 100 kilogray applied at a power of 300 kW.

Electron beam emitters are devices with a scanning beam with a sector width of 1 meter, corresponding to the width of the conveyor belt. In some embodiments, electron beam emitters with a large fixed beam width are used.

Factors including belt/beam width, desired dose, biomass feedstock density, and applied power govern the number of electron beam emitters required for a plant to process 2,000 tons of dry feedstock per day.

Treatment with ultrasound because before applying ultrasonic energy, the irradiated biomass raw material is mixed with water to obtain a suspension. A separate sonication system may exist for each electron beam feedstock stream, or multiple electron beam streams may be collected as feedstock for a single sonication system.

In each sonication system, irradiated biomass feedstock is fed into vessel 1214 through first inlet port 1232, and water is fed into vessel 1214 through second inlet port 1234. Appropriate valves (manual or automatic) control biomass feedstock flow and water flow to obtain the desired feedstock ratio. biomass and water (for example, 10 95 cellulosic material, mass by volume). Each container 1214 includes a mixer 1240 for mixing the contents of the volume 1236 and dispersing the raw biomass in water.

In each sonication system, the slurry is pumped (eg, using a vortex impeller pump 1218) from a vessel 1214 into and through a flow cell 1224 that includes an ultrasonic transducer 1226. In some embodiments, the pump 1218 is adapted to agitate the slurry 1216 so that the mixture of biomass feedstock and water was essentially homogeneous in the inlet channel 1220 of the flow cell 1224. For example, the pump 1218 may agitate the slurry 1216, creating a turbulent flow that is maintained throughout the conduit between the first pump and the inlet channel 1220 of the flow cell 1224.

In the flow cell 1224, an ultrasonic transducer 1226 transmits ultrasonic energy to the slurry 1216 as the slurry flows through the flow cell 1224.

Ultrasonic transducer 1226 converts electrical energy into high-frequency mechanical energy (e.g., ultrasonic energy), which is then delivered to the suspension via booster 48. Ultrasonic transducers (e.g., from Nieuwspeg OA, Ips. oi Kipdugos,

Mem Uegzeu) capable of delivering a constant power of 16 kilowatts are commercially available.

Ultrasonic energy passing through the booster 1248 in the reactor volume 1244 creates a series of compressions and vacuums in the process stream 1216 with an intensity sufficient to cause cavitation in the process stream 1216. The cavitation disaggregates the components of the biomass feedstock, including, for example, cellulosic and lignocellulosic material dispersed in the process stream 1216 (eg, suspensions). Cavitation also leads to the formation of free radicals in process water 1216 (eg

From the suspension). These free radicals act to further degrade the cellulosic material in process stream 1216. Typically, approximately 250 MJ/m3 of ultrasonic energy is applied to process stream 1216 containing fragments of poplar sawdust. Other levels of ultrasonic energy (from about 5 to about 4000 MJ/m3, such as 10, 25, 50, 100, 250, 500, 750, 1000, 2000, or 3000 MJ/mU) can be applied to other biomass feedstocks. After exposure to ultrasonic energy in the volume of the reactor 1244, the process flow 1216 leaves the flow cell 24 through the outlet channel 1222.

The flow cell 1224 also includes a heat exchanger 1246 in thermal contact with at least a portion of the reactor volume 1244. A cooling fluid 1248 (e.g., water) enters the heat exchanger 1246 and absorbs the heat generated when the process stream 1216 (e.g., slurry) is irradiated into the reactor vessel 1244. In some embodiments, the flow of cooling fluid 1248 entering the heat exchanger 1246 is controlled to maintain an approximately constant temperature in the reactor vessel 1244. Additionally or alternatively, the temperature of the cooling fluid 1248 entering the heat exchanger 1246 is controlled to maintain an approximately constant temperature in the reactor volume 1244.

The outlet channel 1242 of the flow cell 1224 is located near the bottom of the container 1214 to induce gravity flow of the process flow 1216 (e.g., slurry) from the container 1214 toward the inlet channel of the second pump 1230, which pumps the process flow 1216 (e.g., slurry) toward the subsystem basic processing.

Sonication systems may include a single flow path (as described above) or multiple parallel flow paths, each associated with individual sonication elements. Multiple sonication elements can also be arranged in series to increase the irradiation energy applied to the slurry.

Basic processing

Before fermentation, a vacuum filter with a rotating drum removes solid particles from the suspension. The liquid from the filter is pumped out in a cooled state before entering the fermenters. The filtered solids pass through the downstream processing subsystem for further processing.

Fermentation tanks are large stainless steel tanks, under low 60 pressure, with conical bottoms and slow stirrers. Several containers for the first stage of fermentation can be arranged in series. The temperature in the containers for the first stage of fermentation is regulated to 30 "C using external heat exchangers. Yeast is added to the container for the first stage of fermentation at the beginning of each series of containers, and it passes through other containers located in a row.

The second stage of fermentation consists of two consecutive continuous fermenters.

Both fermenters are constantly stirred with slow mechanical stirrers. The temperature is controlled by chilled water in internal heat exchangers with constant recirculation.

Recirculating pumps are pumps with moving cavities because the solids concentration is high.

The gases leaving the fermentation tanks and fermenters are combined and washed in a column with a counterflow of water before being released into the atmosphere. The off-gases are scrubbed to extract ethanol, not to control emissions to the atmosphere.

Further processing

Distillation

Distillation and adsorption on molecular sieves are used to extract ethanol from raw fermentation broth and obtain 99.5% ethanol. Distillation is carried out in two columns: the first, called the fermentation column, removes dissolved COg and most of the water, and the second concentrates ethanol almost to an azeotropic mixture.

All water from the practically azeotropic mixture is removed by adsorption on vapor phase molecular sieves. Regeneration of adsorption columns requires that the ethanol-water mixture be recycled to the distillation for separation.

Fermentation off-gases (which contain mainly CO2, but also some ethanol) and off-gas from the fermentation column are cleaned in a water scrubber, which removes almost all the ethanol. The gas leaving the scrubber is fed to the first column for distillation together with the fermentation broth.

The sediment after the first distillation contains all unconverted insoluble and dissolved solids. Insoluble solids are dewatered by a pressure filter and sent to the combustion chamber. The liquid from the pressure filter, which is not reused, is concentrated in a multibody evaporator using

From waste heat from distillation. The concentrated syrup from the evaporator is mixed with the solids sent to the combustor, and the evaporator condensate is used as relatively clean recycle water for processing.

Since the amount of distillation water that can be recirculated is limited, an evaporator is included in the process. The total amount of water from the pressure filter that is directly recirculated is set at 25 95. This stream contains organic salts such as ammonium acetate or lactate, accumulated fluid components that are not disposed of by the body, or inorganic compounds in the biomass. Recirculation of very large amounts of this material can lead to levels of ionic strength and osmotic pressure that can be detrimental to the efficiency of the fermenting organism. For water that is not recirculated, the evaporator concentrates the dissolved solids into a syrup that can be sent to the combustion chamber, minimizing the burden on waste water treatment.

Treatment of waste water

The waste water treatment department treats process water for reuse to reduce the plant's additional water needs. Waste water is first filtered to remove large particles, which are collected in a waste container and sent to a landfill. After screening, anaerobic digestion and aerobic digestion are performed to break down the organic material in the stream. Anaerobic digestion results in a methane-rich biogas stream that is fed into the combustion chamber. Aerobic digestion results in a relatively clean water stream for reuse in the process, as well as a bottom that is primarily composed of cell mass. The sludge is also burned in the combustion chamber. This screening/anaerobic digestion/aerobic digestion scheme is standard in today's ethanol industry, and equipment in the 1-5 million gallons per day range can be obtained from suppliers as "off-the-shelf" units.

Combustion chamber, steam boiler and turbogenerator

The purpose of the combustion chamber, boiler and turbogenerator system is to burn various by-product streams to generate steam and electricity. For example, some part of lignin, cellulose and hemicellulose remains unconverted during pretreatment and basic processes. Most of the waste water from the process is concentrated into a syrup with a high 60 content of soluble solids. Anaerobic digestion of the remaining waste water leads to the formation of biogas with a high methane content. Aerobic digestion leads to a small amount of waste biomass (bottom). Combustion of these by-product streams to generate steam and electricity allows the plant to be energy independent, reduces the cost of solid waste disposal, and provides additional revenue through the sale of excess electricity.

Three streams of primary fuel (solids after distillation, biogas and evaporator syrup) are fed into the combustion chamber with a circulating fluidized bed. A small amount of waste biomass (sludge) after the treatment of waste water is also sent to the combustion chamber.

The fan supplies air to the combustion chamber. Treated water enters the circuit of the heat exchanger in the combustion chamber, evaporates and is subjected to excessive heating to steam at 510 "С (950 "РЕ) and 86 atm. (1265 psi abs. (8.7 MPa)). Furnace gases from the combustion chamber preheat the air entering the combustion chamber and then penetrate a fabric filter to remove particles that are sent to landfill. The gas exits through the chimney.

A multi-stage turbine is used to generate electricity. Steam comes out of the turbine in three different states for injection into the reactor for pretreatment and heat exchange during distillation and evaporation. The other steam is condensed with cooling water and returned to the boiler feed water system along with condensate from the various heat exchangers in the process. Water from a well is used as additional water to replace the flow used in direct injection.

Example 21 - Production of feed for animals from millet

A 1,500 lb (680 kg) millet slip is purchased from the farm and transported to the processing site. The material is fed into the Z Pr Ripsp Wacdy shredder at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material.

Zo The candy-like material is fed into a Mipsop rotary knife cutter, model 50230. The discharge screen has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage.

Cows are fed the required amount of pellets per day.

Example 22 - Production of feed for animals from millet

A 1,500 lb (680 kg) millet slip is purchased from the farm and transported to the processing site. The material is fed into the Z Pr Ripsp Wacdy shredder at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material.

The candy-like material is fed into a cutting device with a rotating knife Mipsop, model

ZS30. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used. (510)

Table 10

Parameters of KpodoihopF TT 200 bundles of SG! nominal (maximum): 10 MeV (10 keV - 250 keV)

Energy dissipation at 10 MeV Full width at half maximum (EMUNM) 300 keV

Beam power at 10 MeV Guaranteed operating range from 1 to 80 kW (Power consumption 7777711

OHM):

With a beam power of 50 kW -210 kW

With a beam power of 80 kW -260 kW

SystemavE 11111111 107.5:1 MHz

Tpotvop TN7VI

Scanning horn pnn"'nnshshshshshshshshshhn

Nominal scan length (measured at | 120 cm 25-35 cm from the window - 077 - t scan

Nominal scan frequency (at 100 Hz and 5 95 maximum scan length)

Uniformity of scanning (over 90 Fo 59 of the nominal scan length) o

Table 11

Dosages delivered to samples

Total dosage (Mrad)

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 23 - Production of fodder for animals from alfalfa

A 1,500 lb. (680 kg) bale of alfalfa is purchased from the farm and transported to the processing site. The material is fed into the Z Pr Ripsp Vatsodi shredder at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material.

The candy-like material is fed into a Mipsop rotary knife cutter, model 50230. The discharge screen has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 24 - Production of fodder for animals from alfalfa

A 1,500 lb. (680 kg) bale of alfalfa is purchased from the farm and transported to the processing site. The material is fed into the Z Pr Riipsp Vacoi shredder at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material.

The candy-like material is fed into a Mipsop rotary knife cutter, model 50230. The discharge screen has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

These processed samples are compacted to form pellets suitable for consumption by cows

Zo and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 25 - Obtaining feed for animals from paper

A 1,500 pound (680 kg) sheet of paper is folded flat and fed into the Z Pr Ripspj Vatsop shredder at a rate of approximately 15 to 20 pounds per hour (6.8-9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm).

The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The candy-like material is fed into a Mipsop model 5230 rotary knife cutter. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 43:1.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 26 - Obtaining feed for animals from paper

A 1,500 pound (680 kg) sheet of paper is folded flat and fed into the Z Pr Ripspj Vatsop shredder at a rate of approximately 15 to 20 pounds per hour (6.8-9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm).

The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The candy-like material is fed into a Mipsop rotary knife cutter, model 5230. The discharge screen has holes of 1/8 inch (0.32 cm). The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 27 - Obtaining fodder for animals from grass

A 1,500-pound (680-kg) wad of grass is fed into the shredder Z Pr Riipsp

Water at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The candy-like material is fed into a Mipsop rotary knife cutter, model 53230. The discharge screen has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm).

A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 431.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

From Example 28 - Obtaining fodder for animals from grass

A 1,500-pound (680-kg) wad of grass is fed into the shredder Z Pr Riipsp

Water at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The candy-like material is fed into a Mipsop rotary knife cutter, model 53230. The discharge screen has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm).

A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 29 - Production of animal feed from wheat straw

A 1,500 lb (680 kg) load of wheat straw is fed into a shredder

From pr Ripsp Vatsoi at a rate of about 15 to 20 pounds per hour (6.8-9.1 kg/hr.). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and thick, because it is equivalent to the thickness of the original material. The candy-like material is fed into a Mippop Model 5230 Rotary Knife Cutter. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 4371.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 30 - Production of animal feed from wheat straw

A 1,500 lb (680 kg) load of wheat straw is fed into a shredder

From pr Ripsp Vatsoi at a rate of about 15 to 20 pounds per hour (6.8-9.1 kg/hr.). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The candy-like material is fed into a Mippop Model 5230 Rotary Knife Cutter. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

These processed samples are compacted to form pellets suitable for consumption by cows

Zo and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 31 - Production of feed for animals from biomass

1,500 lb (680 kg) millet, alfalfa, paper, grass, and wheat straw straws are individually fed into the Z Pr Riipsp Vatsodi shredder at a rate of approximately 15 to 20 lb/hr (6.8-9.1 kg/hr .). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The confetti-like material is fed into a cutting device with a rotary knife Mypbop, model 5C30.

The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 32 - Production of feed for animals from biomass

1,500 lb (680 kg) millet, alfalfa, paper, grass, and wheat straw straws are individually fed into the Z Pr Riipsp Vatsodi shredder at a rate of approximately 15 to 20 lb/hr (6.8-9.1 kg/hr .). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. Similar to confetti because the material is fed into a cutting device with a rotary knife Mypbop, model 5C30.

The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 33 - Production of feed for animals from biomass

A 1,500 lb (680 kg) millet, alfalfa, paper, grass, and wheat straw straw is mixed and fed into a Z Pr Riipsp Vatsody shredder at a rate of approximately 15 to 20 lb per hour (6.8-9.1 kg/hr .). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The confetti-like material is fed into a cutting device with a rotary knife Mypbop, model 5С30.

The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 34 - Production of feed for animals from biomass

1500 lb (680 kg) millet, alfalfa, paper, grass, and wheat straw straws are mixed and fed into the Z Pr Riipsp Vatsodyi shredder at a rate of approximately 15 to 20 lb/hr (6.8-9.1 kg/hr .). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The confetti-like material is fed into a cutting device with a rotary knife Mypbop, model 5C30.

The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average GG /O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF T1200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

These processed samples are compacted to form pellets suitable for consumption by cows and other livestock. Pellets are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 35 - Production of feed for animals from biomass

A 1,500 lb (680 kg) millet, alfalfa, paper, grass, and wheat straw straw is mixed and fed into a Z Pr Riipsp Vatsody shredder at a rate of approximately 15 to 20 lb per hour (6.8-9.1 kg/hr .). The shredder is equipped with two 12- to 60-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm)

unloading sieve. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The confetti-like material is fed into a cutting device with a rotary knife Mypbop, model 5C30.

The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average of 1/0 431.

Processed samples are combined with dry barda (000) to obtain a mixture suitable for consumption by cows and other livestock. These mixtures are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 36 - Production of feed for animals from biomass

A 1,500 lb (680 kg) millet, alfalfa, paper, grass, and wheat straw straw is mixed and fed into a Z Pr Riipsp Vatsody shredder at a rate of approximately 15 to 20 lb per hour (6.8-9.1 kg/hr .). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The confetti-like material is fed into a cutting device with a rotary knife Mypbop, model 5С30.

The discharge screen has 1/8 inch (0.32 cm) holes. The gap between the rotating and fixed blades is set at approximately 0.020 inches (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average G/O of 43:1.

Samples are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

Processed samples are combined with dry barda (005) to obtain a mixture suitable for consumption by cows and other livestock. These mixtures are distributed on farms and stored in silos for storage. These pellets are fed to cows and other livestock.

Example 37 - Autonomous agriculture

A farmer harvests millet and sends it to a processing plant for processing.

The millet is processed as described in Example 21. The processed material is supplied to the farmer in pellet form, which is fed to his cows and other livestock.

Example 38 - Autonomous agriculture

A farmer harvests millet and sends it to a processing plant for processing.

The millet is processed as described in Example 22. The processed material is supplied to the farmer in the form of pellets, which are fed to his cows and other livestock.

Example 39 - Autonomous agriculture

A farmer harvests millet and processes it using equipment located on the farm. The millet is processed as described in Example 21. The processed material is supplied to the farmer in pellet form, which is fed to his cows and

BO other cattle.

Example 40 - Autonomous agriculture

A farmer harvests millet and processes it using equipment located on the farm. The millet is processed as described in Example 22. The processed material is supplied to the farmer in the form of pellets, which are fed to his cows and other livestock.

Example 41 - Fermentation in a rotary flask using Rispia 5iiryov

Brief summary

Fermentation in a rotary flask using different enzymes and Rispia 5iiryi5 was carried out with the use of physical treatment. because the Protocol

The experiments were performed according to the parameters listed in Table 12.

Table 12

Chemical substances and materials used for the experiment in a rotating flask

Obtaining seed material

The working cell bank of R. 5iirie MEKI U-7124 is obtained from a rehydrated lyophilized culture obtained from AKZ Siyige SoPesiop. Cryoflacons containing the culture of R. 5Iiriy5 in 15

Fo ob./0b. glycerol, store at -75 "С. Part of the thawed material of the cell bank is applied with strokes to broth for yeast and mold (SUM) "t 20 g/l agar (pH 5.0) and incubated at 30 "С for 2 days. The cups are kept for 2 days at 4 "C before use.

In a 250 mL Erlenmeyer flask containing 100 mL medium (40 g/L glucose, 1.7 g/L yeast nitrogen base, 2.27 g/L urea, 6.56 g/L peptone, 40 g/ l xylose, pH 5.0), inoculate one colony and incubate it for 24 hours at 25 °C and 150 rpm. After growth for 23 hours, samples are taken and analyzed for optical density (600 nm in a UV spectrophotometer) and purity (Gram stain) Based on these results, two seed flasks, each with an optical density (003) of 4 to 8, are pooled to inoculate all the culture flasks.

Illustrative experiments

Experiments are conducted to 1) determine the exact power of the device for ultrasonic treatment and temperature regulation (below 60 "C), and 2) confirm the concentration

SeiPisiavi 1.5 ES and Momogute 188 with and without Riygopis E-68.

Five hundred milliliters of water are added to an analytical beaker with a volume of 1 liter. Horn of the device for irradiation Vgapzop Mode! 450 Bopiegs are placed 1/2 inch (1.25 cm) into a beaker and set on maximum continuous power for 60 minutes. Water temperature is measured every 10 minutes during 60 minutes of ultrasound treatment.

The experiment is carried out to determine: 1) whether the concentration of SePysiabi is 1.5 EC and

Momo7ute 188 (0.5 mL and 0.1 mL per gram of biomass, respectively) is sufficient for spin-flask experiments, and 2) whether the addition of Rishigopis E68 enhances cellulose hydrolysis. Prepare four 250-ml flasks with 100 ml of sterile broth (1.7 g/l yeast nitrogen bases, 2.27 g/l urea, 6.56 g/l peptone, pH 5.0). Duplicate flasks contain 1 95 wt./rev. Riygopis E-68. In the flask

ZoiKkaBios crystalline cellulose (b g) is added and allowed to soak at room temperature for 14 hours. Seysiavi 1.5 EU and Momo7ute 188 (0.5 ml and 1 ml per gram of ZoiKkagios, respectively) are added and each flask is incubated at 50 °C for 24 hours at 100 rpm.

Taking samples from all four flasks is carried out before the addition of the enzyme and 24 hours after the addition of the enzyme and they are analyzed for glucose concentration using the U5I Viospet analyzer (U5I, Ipieg5siepse). One milliliter of Rispia 5iirini seed flask content is added to four flasks and incubated at 25 °C and 125 rpm for 24 hours. Samples are taken from each flask before inoculation and after incubation for 24 hours and analyzed for ethanol concentration using the U5I Viospet analyzer (U5I, Ipteg5siepse).

Tested flasks

Test flasks are 2.8 L Fernbach flasks containing 900 mL of broth (1.7 g/L yeast nitrogenous bases, 2.27 g/L urea, 6.56 g/L peptone, pH 5.0) . Control flasks are 250 mL flasks containing 100 mL of broth (40 g/L glucose, 1.7 g/L yeast nitrogenous bases, 2.27 g/L urea, 6.56 g/L peptone, 40 g/L l xylose, pH 5.0). The exact characteristics of each flask are determined by Huieso and are described in Table 13, below.

The samples are not sterilized before the start of the experiment. All samples are added to flasks and allowed to infuse for 15 hours at room temperature. Some of the samples are sonicated for one hour using the Vgahop Mode sonicator! 450 Zopieg equipped with a 1/2 inch (1.25 cm) disintegrating horn. Initially, it was planned to divide the contents of the flasks into two parts, and each half to be treated with ultrasound continuously at the maximum power of the equipment up to 450 watts (the permissible power depends on the viscosity of the sample) for 1 hour. The power parameter C and the pulse utilization factor of 90 95 were sufficient to mix the contents of the beaker. With the power parameter C, the meter reading is from 30 to 40. The calculated power is 40-60 watts.

Initially, it was planned to mix several samples (see Table 13) for different periods of time using a laboratory homogenizer ROIMTKOM RT 10/35 (or rotor/stator) at 25,000 rpm. during different periods of time. Samples 222 and 223 are divided into two glasses and processed for 30 minutes using a large Kipetaiis Royuigop RT 10/35. The generator (tip) is a PTA 20 with a stator diameter of 20 mm. The device operates at a speed of 11,000 rpm. Work at above 11,000 rpm. leads to splashing of the contents of the glass, displacement of the glass and excessive heating of the equipment. After samples 223 and Zh24

Roiugop RT 10/35 stopped working, presumably due to excessive use with rather viscous samples. Thus, they use a portable Royiop RT1200S.

The generator (tip) is RT-BA 1212 with a stator diameter of 12 mm. The device could operate at 25,000 rpm. The operator noticed that on a portable device at 25,000 rpm. observed a similar degree of agitation compared to the larger model at 11,000 rpm. The operator periodically stirs the sample to ensure uniform mixing.

Samples 19 to 22 are mixed using a portable Roiugop RT 12000.

Enzymatic pre-treatment includes: 1) enzyme complex Ei - AsseyiIegazet 1000 at a loading density of 0.25 ml per gram of substrate; and 2) E2 - SePisiavi 1.5 BO and Momogute 188 at a loading concentration of 0.5 and 0.1 ml per gram of substrate, respectively. After physical pretreatment (see Table 13, below), the appropriate enzyme(s) and flasks are added

This is maintained at 50 °C and 125 rpm for 20 hours. After 20 hours, the flasks are cooled to room temperature for 1 hour before adding R. 5iiiiii5.

Table 13

Generalization of tested treatments

Number Concentration Enzymatic

Test number sample sample Physical processing processing (50 "C, g/900 ml) 21 h.)

Control (250-ml flask), performed in duplicate every week

Impregnation for 15 hours. at c.t. at c.t. at c.t. at c.t.

Orzhiv shen in the case of kt. at c.t.

Continuation of table 13 ren in fizheeennnnia! o in at k.t. ren ovo frreennnniavo at k.t.

Impregnation for 15 hours. 10 ХР-10е 35 at c.t. E2 ultrasound treatment for 1 hour.

Impregnation for 15 hours. 11 KhrR-ZOe 35 at c.t. E2 ultrasound treatment for 1 hour.

Impregnation for 15 hours. 12 KhR-BOe 35 at c.t. E2 ultrasound treatment for 1 hour.

Impregnation for 15 hours. 13 KhR-10e 35 at kt. E2 ultrasound treatment for 10 min.

Impregnation for 15 hours. 14 KhrR-ZOe 35 at c.t. E2 ultrasound treatment for 10 min.

Impregnation for 15 hours. 15 KhR-BOe 35 at c.t. E2 ultrasound treatment for 10 min.

Impregnation for 15 hours. 16 KhR-10e 35 at c.t. E2 ultrasound treatment for 30 min.

Impregnation for 15 hours. 17 KhrR-ZOe 35 at c.t. E2 ultrasound treatment for 30 min.

Impregnation for 15 hours. 18 KhR-BOe 35 at c.t. E2 ultrasound treatment for 30 min. at c.t. 10 min. on the rotor/stator at kt. 10 min. on the rotor/stator at kt. 10 min. on the rotor/stator at kt. 30 min. on the rotor/stator at kt. 30 min. on the rotor/stator at kt. 30 min. on the rotor/stator

Analysis

Samples are taken from each flask after physical and/or enzymatic pretreatment (immediately before the addition of R. viiirii5) and analyzed for glucose concentration using a U5I Viospet analyzer (U5I, Ip(eg5siepse). The samples are centrifuged at 14,000 rpm for 20 minutes and the supernatant is stored at -20 "C. Before analysis, the samples are diluted to a glucose level of 0-25.0 g/L. The glucose standard is analyzed approximately after every 30 samples to ensure that the integrity of the membrane is maintained.

A total of five samples are taken from each flask after 0, 12, 24, 48 and 72 hours and analyzed for ethanol concentration using the U5I Viospet analyzer based on alcohol dehydrogenase analysis (U5I, Ipiegosiepse). The samples are centrifuged at 14,000 rpm. for 20 minutes and the supernatant is stored at -20 "C, and diluted to an ethanol concentration of 0-3 g/l before analysis. A standard in the form of ethanol at a concentration of 20 g/l is analyzed approximately after every 30 samples in order to make sure that that membrane integrity was maintained throughout the assay.

A sample of the seed flask is analyzed to determine the initial concentration of cells in the tested flasks. In addition, one sample is taken from each flask after incubation for 72 hours and analyzed for cell concentration. Appropriately diluted samples are mixed with 0.05 96 trypan blue and applied to a Meicyrateseg hemocytometer. Cells are counted at 40X magnification.

The results

Experiments

The results of the experiment with the ultrasonic treatment device are presented in Table 14.

There were no problems with overheating the water.

Table 14

An experiment with a device for ultrasound treatment

Temperature СС) ов ны и пох С: PO

The results of the experiment to confirm the concentration of SeiPysiavi 1.5 ES and Momo7ute 188 with

Rishgopis E-68 and without it are presented in tables 15 and 16. Cellulose is added to each flask at a concentration of 60 g/l (ZoiIKanRios). After incubation for 24 hours, from 33.7 to 35.7 g/l of glucose is obtained (from 30.3 to 32.1 g/l of split cellulose).

After incubation for 24 hours with R. 5iiryi5, 23.2-25.7 g/l of glucose remained in the flasks. This indicates that not all of the glucose was used during the 24-hour incubation.

There were no data on the toxicity of Riygopis E-68 against P. spp. However, there was no increase in the amount of glucose after treatment with the enzyme for 24 hours when Riigopis E-68 was added.

Table 15

Results for glucose

Glucose concentration (g/l)

Flask Before treatment After treatment with enzyme After R. ziirii5 enzyme 50 "C, 24 hours, 100 rpm | for 24 hours

Which contains Rishgopis V

Table 16

Results for ethanol

Ethanol concentration (g/l) at certain time points (hours)

Flask - -8-- i

O (inoculation, after enzyme treatment) | R. 5iiiii5 within 24 hours.

Which contains Rishgopis A

What does Rishopis contain? | -////7777777771017000777111111111111111111111111117361

During the first week of testing, the seed flask has an optical density (600 nm) of 9.74 and a cell concentration of 4.21x108 cells/ml. Nine ml of inoculum material is added to each of the test flasks and 1 ml is added to the control flasks (1 95 vol./06.). Thus, the initial concentration of cells in each flask is 4.21x105/ml.

During the second week of testing, the seed flask has an optical density (600 nm) of 3.02 and a cell concentration of 2.85x109 cells/ml. In order to account for differences in the number of cells and 00, 12 ml of the inoculum material is added to each of the test flasks and 1.5 ml is added to the control flasks (1.5 95 vol./06.). Thus, the initial concentration of cells in each flask is 3.80x106/ml.

The concentration of ethanol in the flasks is presented in Table 17. The highest concentration of ethanol is observed in flask Ж (Sample ХР, impregnation overnight, E2 treatment at 50 "С for 21 hours). The concentration obtained within 48 hours from the initial 35 grams is 19.5 g /l (17.55 g/per flask).The yield of ethanol (grams of ethanol/grams of substrate) in the flask is 0.50.

Table 17

Ethanol concentration 016 |oily salt |006 77771775... | 0312 | 031 |1030| 1130 | 18.80 7777776 | 0399 | 2 sh073 | 755 | 195507 | 19.00 7771777... .| 0419 | 038 | 473 | 16807 | 15.40 7777778 ..yuyuyuyuyuyu.| 0370 | 046 | 056| 986 | 15.50 77778... | ol83 | 047 | 053| 5 yush 1200 | 140 7.771077 | 0216 | 035 2 sh | bli| 13.80. | 1560 77713... 086 | sh099 2 4 yush |842| 1050 25 DSH YuuKH | 1420 7717717... 0989 | yush045 | 173| 1060 | 12.00 777723. | 096 | yush 057 sh |677| lo | l2lo “Samples were analyzed twice with the same result.

Flasks with an ethanol concentration greater than 15 g/l are highlighted in bold.

The results of the glucose analysis are presented in Table 18. After treatment with the enzyme for 21 hours, the highest concentration of glucose is 19.6 g/L (17.6 grams/per flask) in flask b (sample XP, overnight impregnation, E2 treatment at 50 " C for 21 hours). This flask is also the flask with the highest concentration of ethanol (see Table 17). Very little glucose remained in the flasks after 72 hours. No glucose was detected in flasks 1 and 2.

Table 18

Glucose concentration

Концентрація глюкози (г/л) у визначені моменти часу (год.) 00111111 0001 а111111711111111111111110011111111111171711111111100071 нннижини,ининншсснишшшншишиши у ти 81111111111Г11111111111111154777777771117111171 17717171 006 71 81111118 17111111111009 8 111111111111111111111158777777777777171171177717171717171711006 1 в 11111111111111111111155.77771717171717171717171177171717171717100621щ

The results of direct cell counting are presented in Table 19. The concentration of viable cells was higher than in the control flasks. The lowest number of cells is observed in flasks 1-4.

Table 19

Number of cells 2 411111111111100611111111 61111110 tu n"'nИХХХШШВІХШВВВВЛИИЙНВВВВСТССКСХХХИИИИИИИИИИИЯIVVVVVVVNYA n"niiХkushnnnnshih today

Continuation of table 19

Example 42 - Production of bioconversion products from biomass

A 1,500-pound (680-kg) batch of biomass is fed into the shredder Z Pr Riipsp

Water at a rate of approximately 15 to 20 pounds per hour (6.8 to 9.1 kg/hr). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The candy-like material is fed into a Mipsop model 5230 rotary knife cutter. The discharge screen has 1/8 inch (0.32 cm) holes. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm).

A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 431.

Materials are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

Processed materials are distributed in table-top sterilized Mem/Vhyp5ulisK zsiepiys fermenters in the form of a liquid medium, which is made to support the growth, reproduction and/or activity of a microorganism selected due to its ability to produce the required bioconversion product. Different concentrations of processed materials are added in combination with different amounts of other auxiliary materials, which are usually necessary for the growth, reproduction and or activity of the selected microorganisms. A source of nitrogen is also added to the medium.

The concentration or amount of processed materials and each of the auxiliary materials (including the nitrogen source) is recorded on a laboratory notebook or computer hard disk.

Z The starter culture of the selected microorganism is added to each of the different culture solutions in the fermenters. Each of the inoculated culture solutions is incubated at a temperature from about 15 "C to about 40 "C for 4 to 48 hours under aerobic or anaerobic conditions. After cultivation, microorganisms and cell supernatants are collected and optionally separated using centrifugation. Then the samples are either frozen

C5 for storage, or evaluated to determine the level of bioconversion of the product in cells or supernatant. The results are recorded, and the experiments are repeated until the maximum yield of the bioconversion product is obtained. The culture solution and conditions used to obtain this maximum yield are scaled up for use in large-scale fermentation.

Example 43 - Large-scale production of bioconversion products from biomass

A 1,500 lb (680 kg) batch of biomass is mixed and fed into a shredder

From pr Ripsp Vatsoi at a rate of about 15 to 20 pounds per hour (6.8-9.1 kg/hr.). The shredder is equipped with two 12-inch (30-cm) rotating blades, two fixed blades and a 0.30-inch (0.8-cm) discharge screen. The distance between the rotating and fixed blades is set at 0.10 inches (0.25 cm). The shredding material coming out of the device resembles confetti, 0.1 inch (0.25 cm) to 0.5 inch (1.3 cm) wide, 0.25 inch (0.6 cm) to 1 inches (2.5 cm) and a thickness equivalent to the thickness of the original material. The confetti-like material is fed into a Mippop model 5230 rotary knife cutter. The discharge screen has 1/8 inch (0.32 cm) openings. The clearance between the rotating and fixed blades is set at approximately 0.020 inch (0.05 cm). A rotating knife cutter crushes the confetti-like fragments with the blade tips, breaking the fragments and releasing the fibrous material at a rate of approximately one pound per hour (454 grams per hour). The average fiber length is 1.063 mm and the average fiber width is 0.0245 mm, giving an average I/O of 43:1.

Materials are treated with a beam of electrons using an accelerator located in a storage with non-attenuating waves KpodoihopF TT200, which delivers electrons with an energy of 5

MeV at an output power of 80 kW. Table 10 describes the parameters used. Table 11 describes the nominal dose used.

The processed materials are used to produce the culture solution defined in Example 42. The selected microorganism and the culture solution are combined in a fixed-volume, bulk-fed fermenter and maintained using the conditions and for the time period defined in Example 42. Concentrated Culture Solution , containing recycled materials, is added to the fermenter if necessary. In addition, the bioconversion product and microorganisms are removed from the fermenter and processed for storage or use.

Example 44 - Large-scale production of bioconversion products from biomass using livestock waste as a nitrogen source

Bioconversion products are obtained as described in example 43, using livestock waste as a nitrogen source. Before use, livestock waste is sterilized using filtration or steam and high pressure sterilization. Sterilized livestock waste is dried before adding to the culture solution.

Example 45 - Large-scale production of Ryzagisht mepepaisht (ATSS 20334) from biomass

Eizagicht mepepaisht is cultivated using the method described in example 43. Collected

E. mepepaisht is combined with rehydrated egg white, onions, textured wheat protein (wheat protein, wheat starch) and canola oil and processed for use as human food.

OTHER EXECUTION OPTIONS

A number of variants of implementation of the invention are described. However, it is clear that various modifications can be made without departing from the essence and scope of the invention.

In some embodiments, relatively low doses of radiation, optionally in combination with sonic energy, such as ultrasonic energy, are used to cross-link, graft, or otherwise increase the molecular weight of a natural or synthetic hydrocarbon-containing material, such as any of the materials in any -what form (for example, fibrous form) described in this document, for example shredded or unshredded cellulosic or lignocellulosic materials, such as cellulose. Cross-linking, grafting, or otherwise increasing the molecular weight of a natural or synthetic hydrocarbon-containing material can be accomplished in a controlled or predetermined manner by selecting the type or types of radiation used (eg, e-beam and ultraviolet radiation or e-beam and gamma radiation) and/or dose or number of doses of applied radiation.

For example, a fibrous material that includes a first cellulosic and/or lignocellulosic material having a first molecular weight can be irradiated to produce a second cellulosic and/or lignocellulosic material having a second molecular weight greater than the first molecular weight. For example, if gamma radiation is used as the radiation source, a dose of from about 0.2 Mrad to about 10 Mrad can be used, for example from about 0.5 Mrad to about 7.5 Mrad or from about 2.0 Mrad to about 5.0 Mrad. If e-beam irradiation is used, a lower dose (relative to gamma radiation) can be used, such as from about 0.1 Mrad to about 5 Mrad, for example from about 0.2 Mrad to about 3 Mrad or from about 0.25 Mrad to about 2.5 Mrad.

One of the following additives may be added to fibrous materials, compacted fibrous materials, or any other materials and composites described in this document. Additives can be added, for example, in the form of a solid, liquid or gas. Additives include fillers such as calcium carbonate, silicon dioxide and talc; inorganic flame retardants, such as aluminum oxide trihydrate or magnesium hydroxide; organic flame retardants, such as chlorinated and brominated organic compounds. Other additives include lignin, 60 fragrances, compatibilizers, processing additives, antioxidants, opacifiers,

heat stabilizers, dyes, foaming agents, polymers, such as degraded polymers, photostabilizers, biocides and antistatic agents, such as stearates or ethoxylated fatty acid amines. Suitable antistatic compounds include conductive carbons, carbon fiber, metal fillers, cationic compounds such as quaternary ammonium compounds such as M-(3-chloro-2-hydroxypropyl)/trimethylammonium chloride, alkanolamides and amines. Illustrative degraded polymers include polyhydroxy acids, such as polylactides, polyglycolides and copolymers of lactic acid and glycolic acid, poly(hydroxybutyric acid), poly(hydroxyvaleric acid), lactide and e-caprolactone copolymer, glycolide and e-caprolactone copolymer, polycarbonates, poly(amino acids) , poly(hydroxyalkanoates), polyanhydrides, complex polyorthoesters and mixtures of these polymers.

When the additives described are included, they may be present in amounts calculated on a dry weight basis ranging from less than 1.95% to as much as 80.95% by weight of the total fibrous material. More specifically, the amounts range from about 0.5 wt. 95 to about 50 wt. 95 such as 5, 10, 20, 30 95 or more such as 40 95.

Any additives described herein may be encapsulated, such as spray-dried, or microencapsulated, such as to protect the additives from heat or moisture during storage.

Fibrous materials, densified fibrous materials, resins or additives can be colored. For example, fibrous material can be dyed before being combined with resin and mixed to produce composites. In some embodiments, this coloring may be useful in masking or concealing fibrous material, especially large agglomerates of fibrous material, in molded or extruded parts when desired. Such large agglomerates, when present in relatively high concentrations, may appear as grains on the surfaces of molded or extruded parts.

For example, the desired fibrous material can be dyed using an acid dye, a direct dye, or a reactive dye. Such dyes are available from

Zresiga Juue5, Keagtu, MOU or Keusiope Apiipe Sogrogayop, Spisado, IS. Specific examples of dyes include ZHRESTEAM TSOYANT MEP OMU/ 20, BRESTVASIUTM MEP OMU 401. SOM 200,

I'm getting married! "M VOBAMIME 8, 5RESTVAMMUI!"M MEOTEAI RBEO V, 5RESTEAMIME"M

From VEM2OREVRYOVIME, 5RESTAVASIALO"M VIASK OV, 5RESTAVAMIME"M TORBOTCOYI5E O and

ZRESTAAMIME"M SEEM AND MI. 200 95, each of which is available from Zresig Yuuev.

In some embodiments, concentrates of dyes in resins containing pigments are mixed with dyes. When such mixtures are then mixed with the desired amount of fibrous material, the fibrous material may be colored by the mixing process. Dye concentrates are available from Siagiapi.

The addition of a flavoring or perfuming agent to the fibrous material, densified fibrous material or composites may be preferred.

Mobile processing of biomass

Stationary plants for biomass processing are described. However, depending on the source of the biomass feedstock and the products produced from it, it may be preferable to process the biomass in mobile installations that may be located near the source of the raw material and/or near the markets for the products obtained from the raw material. As an example, in some embodiments, different grasses, such as millet, are used as biomass raw materials. Transporting large volumes of millet from growing areas to processing facilities hundreds or even thousands of miles away can be both energy-intensive and economically expensive (for example, the cost of transporting raw materials by train is estimated to be between 53.00 and 56.00 per a ton per 500 miles). Moreover, some of the millet feedstock products may be suitable for markets in areas where biomass feedstocks are grown (e.g. ruminant feed). Likewise, transporting ruminant feed hundreds or even thousands of miles to market may not be economically feasible.

Thus, in some embodiments, the recycling systems described herein are implemented as mobile reconfigurable recycling facilities. One of the variants of the implementation of such a mobile installation is shown in Fig. 41. Processing plant 8000 includes five transport trucks 8002, 8004, 8006, 8008 and 8010 (although Figure 41 shows five trucks, generally any number of trucks can be used). The 8002 truck includes a water source and recycling system and electrical power systems for other trucks. Each of the trucks 8004, 8006, 8008 and 8010 is adapted for parallel processing of biomass raw materials. 60 The truck 8002 includes a water source inlet 8012 for receiving water from a permanent source (such as a water main) or a container (eg, a container on another truck, or a container or other reservoir located in the processing area). Process water is circulated to each of the trucks 8004, 8006, 8008, and 8010 through water supply pipes 8020. Each of the trucks 8004, 8006, 8008, and 8010 includes a portion of the water pipe 8020. When the trucks are positioned next to each other to form a mobile recycling unit, the portions of the water pipe 8020 connect to form a continuous water conduit.

Each of the trucks 8004, 8006, 8008, and 8010 includes a water inlet 8022 for supplying process water, and a water outlet 8024 for removing process water. Water outlets 8024 in each of the trucks 8004, 8006, 8008 and 8010 lead to a fragmented continuous water discharge conduit 8026, which is similarly connected to a continuous conduit when the trucks are positioned side by side. The spent process water circulates to a water treatment device 8028 in the truck 8002, which treats the water to remove harmful waste materials and then recycles the treated water through the water pipe 8030 back to the feed water pipe 8020. The waste materials removed from the used process water, may be disposed of off-site or may be stored (eg, in another truck, not shown) and transported to a storage facility.

Truck 8002 also includes an electrical supply station 8016 that provides electrical power to each of trucks 8004, 8006, 8008, and 8010.

The electrical power station 8016 may be coupled to an external power source via connection 8014. Alternatively or additionally, the electrical power station may be adapted to generate power (eg, by burning a fuel source).

Electrical power is supplied to each of the trucks 8004, 8006, 8008, and 8010 via an electrical power cable 8040. Each of the trucks 8004, 8006, 8008, and 8010 includes an electrical power terminal 8018 to which devices on the truck that require electrical power are connected. .

Each of the trucks 8004, 8006, 8008 and 8010 includes an inlet 8042 for raw materials and an outlet 8044 for waste. The biomass feedstock enters each of the trucks 8004, 8006, 8008, and 8010 through inlet 8042, where it is processed according to the methods described herein. After processing, the waste material is removed through an outlet 8044. Alternatively, in some embodiments, each of the trucks 8004, 8006, 8008, and 8010 may be connected to a common feed inlet (eg, located in truck 8002), and each truck the vehicle may dump waste material through a common outlet (eg, also located in truck 8002).

Trucks 8004, 8006, 8008 and 8010 can each include different types of recycling modules; for example, on the configuration presented in Fig. 41, each of the trucks 8004, 8006, 8008, and 8010 includes an ion accelerator 8032 (e.g., a peletron-based horizontal tandem bent accelerator), a heating/pyrolysis station 8034, a wet chemical processing module 8036, and a biological processing module 8038. Typically , each of the trucks 8004, 8006, 8008, and 8010 may incorporate any of the recycling systems described herein. In certain embodiments, each of the trucks 8004, 8006, 8008, and 8010 includes identical recycling systems. However, in some embodiments, one or more trucks may have different recycling systems.

In addition, some or all trucks may have certain on-board recycling systems that are not used, depending on the nature of the source material. As a general rule, the layout of the various on-board recycling systems on each of the 8004, 8006, 8008 and 8010 trucks is reconfigurable according to the type of material being processed.

Processing plant 8000 is an illustrative parallel processing plant; each of the trucks 8004, 8006, 8008 and 8010 processes biomass raw materials in parallel. In certain embodiments, mobile recycling facilities are implemented as sequential recycling facilities. One of the variants of the implementation of a sequential mobile processing plant 8500 is presented in fig. 42. Processing plant 8500 includes three railcars 8502, 8504 and 8506 (generally any number of railcars can be used), each of which is adapted to carry out one or more processing stages in the entire biomass processing process. Rail car 8502 includes a raw material inlet for feeding raw material from storage (eg, a storage building or other rail car). Raw materials are transported from one processing module to another,

among three railway cars through a continuous conveyor system. Railcar 8502 also includes an electrical power station 8514 to supply electrical power to each of railcars 8502, 8504, and 8506.

The railway car 8502 includes a device for coarse mechanical processing 8516 and a device for fine mechanical processing 8518 to convert raw materials into finely divided fibrous material. A third mechanical processing device 8520 rolls the fibrous material into a flat continuous bed. The layer of fibrous material is then transported to an ion accelerator 8522 on rail car 8504, in which the fibrous material is exposed to the ion beam. After treatment with an ion beam, the fibrous material is transported to a low-energy electron accelerator 8524.

The fibrous material is then transported to the chemical processing module 8526 in rail car 8506 for one or more stages of chemical processing. Rail car 8506 includes a process water inlet 8532 into which process water is supplied from an external reservoir (eg, a tank or another rail car).

After chemical treatment in processing module 8526, the material is transported to biological processing module 8528 to initiate fermentation of the sugars released from the material. After the biological processing is complete, the material is transported to separator 8530, which diverts useful products to pipeline 8510 and waste materials to pipeline 8512.

Conduit 8510 can be connected to a storage module (for example, a tank car or an external storage container). Similarly, the waste products may be transported via pipeline 8512 to a storage module, such as an ilabo tank car, to an external storage facility. Separator 8530 also recycles clean process water for further delivery to chemical processing module 8536 and/or biological processing module 8528 .

As discussed above, the processing plant 8500 is an example of a sequential configuration of a mobile processing plant; each of the rail cars 8502, 8504 and 8506 includes an excellent set of recycling systems; and the technological flow of raw materials from each car is connected to the next car in sequence to complete the processing sequence.

As a rule, many different types can be used to process biomass raw materials

From processing configurations. Processing plants, both truck-based and train-based, can be adapted for both sequential and parallel operation. As a rule, the arrangement of different processing modules is configurable, and not all processing modules may be used for a particular raw material. When a specific processing module is not used for a specific raw material, the processing module can be diverted from the process stream. Alternatively, the processing module can remain in the general process flow, but it can be disabled so that the raw material passes through the disabled module quickly, without modification.

Mobile processing units may include one or more electronic control devices that automate some or all aspects of the biomass processing process and/or the setup of the mobile unit. For example, the electronic control device may be adapted to receive input information about the feedstock material to be processed and may generate various output information, including a proposed configuration of the mobile processing plant and/or values for one or more processing parameters included in the processing process. biomass to be realized.

Although the transportation by truck is described above, part of the processing plant or the entire processing plant can be transported by any other means, such as by rail or on a marine vessel, such as a ship, barge, boat, dock or floating platform. Also, transportation can be carried out using more than one mode of transport, for example, using a container, both on a ship and on a tractor with a trailer or a train.

In some embodiments, the methods described herein can be performed using, for example, coal (eg, charcoal).

Thus, other implementation options are within the scope of the claims presented below.

Claims (15)

FORMULA OF THE INVENTION 1. The method of preparing feed material, consisting of: changing the molecular structure of polysaccharides of biomass, which contains polysaccharides in the form of cellulose, hemicellulose or starch, by irradiating the biomass with radiation with a dose of at least 5.0 Mrad, using an electron beam with a power of at least 5 kW, to obtain feed material having a number average molecular weight of from about 3,000 Daltons to about 50,000 Daltons, wherein the feed material has nutrient availability that exceeds the nutrient availability of biomass, wherein said method does not include the use of microorganisms. 2. The method according to claim 1, where the feed material is a feed material suitable for animal consumption. 3. The method according to claim 1, where the feed material is feed material for use in agriculture. 4. The method according to claim 1, where the feed material additionally contains an enzyme. 5. The method according to claim 1, where the electron beam has a power of at least 10 kW. 6. The method according to claim 1, where the electron beam has a power of at least 20 kW. 7. The method according to claim 1, where the electron beam has a power of at least 50 kW. 8. The method according to claim 1, where the electron beam has a power of at least 100 kW. 9. The method according to claim 1, in which biomass is exposed to electron beam irradiation at a dose rate of more than 1 Mrad/s. 10. The method according to claim 9, in which the dose rate is from about 1 Mrad/s to about 10 Mrad/s. 11. The method according to claim 1, in which the biomass is irradiated using more than one electron beam irradiation device. 12. The method according to claim 11, which additionally includes the passage of biomass through and between devices for irradiation with an electron beam. 13. The method according to claim 1, in which the biomass contains lignocellulosic material. 14. The method according to claim 13, in which the lignocellulosic material is selected from the group consisting of grasses, rice husks, cake, jute, yarn, flax, bamboo, sisal, abaca, straw, corn cob cores, corn straw, alfalfa, hay, coconut fibers, cotton, sea grass, algae and their mixtures. 15. The method according to claim 14, where the specified material contains cores of corn cobs and/or corn straw. 16. 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