OA16507A - Processing biomass. - Google Patents

Abstract

Feedstocks, obtained at least in part from a plant material that has been modified with respect to its wild type, are processed to produce useful intermediates and products, such as energy, fuels, foods or materials. For example, systems are described that can treat such feedstock materials, e.g., to reduce the recalcitrance of the feedstock, and use the treated feedstock materials to produce an intermediate or product, e.g., by saccharification and/or fermentation.

Description

PROCESSING BIOMASS

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application Serial No. 61/442,781, filed February 14, 2011. The complété disclosure of this provisional application is hereby incorporated by reference herein.

BACKGROUND

Cellulosic and lignocellulosic materials are produced, processed, and used in large quantifies in a number of applications. Often such materials are used once, and then discarded as waste, or are simply considcred to bc waste materials, e.g., bagasse, sawdust, and stover. In some cases, cellulosic and lignocellulosic materials are obtained by growing and harvesting plants.

SUMMARY

Generally, this invention relates to using and/or processing feedstock materials e.g., cellulosic and/or lignocellulosic feedstock materials, including plants that have been modified with respect to their wild types, e.g., genetically modified plants, and to intermedtates and products made therefrom. Many of the methods described herein provide materials that can be more readily utilized by a variety of microorganisms to produce useful intermediates and products, e.g., energy, a fuel, a food or a material. In one aspect, the invention features methods for making products that include physically treating a cellulosic, lignocellulosic and/or starchy feedstock obtained at least in part from a plant that has been modified with respect to a wild type variety of the plant e.g., the plant has been genetically modified. In some embodiments the entire plant can bc used. In certain embodiments, a portion of the plant is utilized.

Some implémentations include one or more of the following features. The feedstock may include a plant that has recombinant DNA and/or recombinant genes. The 25 modified plant may express one or more recombinant materials, for cxamplc, a protein, a polymer and/or a macromolecule. The method may further include obtaining from the feedstock materials such as pharmaceuticals, nutriceuticals, proteins, fats, vitamine, oils,

fiber, minerais, sugars, carbohydrates and alcohols. The feedstock can include a crop residue e.g., com cobs and/or corn stover, wheat straw, or the feedstock can be a genetically modified com, wheat or soybean plant. The method may further include treating the feedstock with an organism and/or enzyme, in some cases producing a sugar e.g., in the form of a solution or suspension. Optionally the sugar can be fermented. The physical treatment can include irradiation of the feedstock. In some implémentations, the irradiated feedstock may be utilized as an edible material, e.g., as an animal feed. If desired, an enzyme such as a cellulase can be added to the edible material, e.g., to increase the nutrient value release.

Irradiating may in some cases be performed using one or more électron beam devices. In some cases, irradiating comprises applying a total dose of from about 5 Mrad to about 50 Mrad of radiation to the feedstock. Irradiation can sterilize the material prior to further processing and or storage prior to use. In preferred implémentations, irradiating reduces the recalcitrance of the feedstock.

The plant may hâve been modified, for example, with a modification including enhancement of résistance to insects, fungal diseases, and other pests and diseasc-causing agents; increased tolérance to herbicides; increased drought résistance; extended température range; enhaneed tolérance to poor soil; enhaneed stability or shclf-lifc; greater yield; Iarger fruit size; stronger stalks; enhaneed shatter résistance; reduced time to crop maturity; more uniform germination fîmes; higher or modified starch production; enhaneed nutrient production, such as enhaneed, steroid, sterol, hormone, fatty acid, glycerol, polyhydroxyalkanoate, amino acid, vitamin and/or protein production; modified lignin content; enhaneed cellulose, hemicellulose and/or lignin dégradation; including of a phenotype marker to allow qualitative détection; reduced recalcitrance and enhaneed phytate metabolism. The plant may be, for example, a genetically modified alfalfa, potato, beet, com, wheat, cotton, rapeseed, rice, or sugarcane plant. The feedstock may include a crop residue from a modified plant, for example the feedstock may include com cobs and/or com stover. The plant may be, for examplc, a genetically modified com or soybean plant, or any of the many genetically modified plants that are grown.

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In another aspect, the invention features a product comprising sugar derived from a feedstock obtained at least in part from a plant that has been modifîed with respect to a wild type variety of the plant, for example the plant has been genetically modifîed.

In a further aspect, the invention features a product comprising an irradiated cellulosic or lignocellulosic feedstock obtained at least in part from a plant that has been modifîed with respect to a wild type variety of the plant. The product may further include a microorganism and/or an enzyme, and in some cases a liquid medium.

Without being bound by any theory, it is believed that the use of modifîed plants can be advantageous over the non-modified wild type. For example, an enhancement of résistance to insects, fungal diseases, and other pests and disease-causing agents; an increased tolérance to herbicides; increased drought résistance; an extended température range; enhaneed tolérance to poor soil; a larger fruit size; stronger stalks; enhanced shatter résistance; reduced time to crop maturity; more uniform germination times; can provide higher yields and a more varied feedstock source, both of which can lower the biomass feedstock cost. In another example, enhanced stability or shelf-lîfe can be advantageous to biomass inventory quality. As another example, enhanced nutrient production, such as enhanced steroid, stcrol, hormone, fatty acid, glycerol, polyhydroxyalkanoatc, amino acid, vitamin and/or protein production can providc products or intermediates with higher nutrient quality that may improve a process e.g., a fermentation, or a product, e.g., an animal feed. Furthermore, for example, higher or modifîed starch production, modifîed lignin content; and/or enhanced cellulose, hemicellulose and/or lignin dégradation can reduce the recalcitrance of the feedstock making it easier to process.

The term “plant,” as used herein, refers to any of various photosynthetic, cukaryotîc, multicellular organisms of the kingdom Plantae, including but not limited to agricultural crops, trees, grasses, and algae.

“Structurally modifying” a feedstock, as that phrase is used herein, means changing the molecular structure of the feedstock in any way, including the chemical bonding arrangement, crystalline structure, or conformation of the feedstock. The change may be, for example, a change in the integrity of the crystalline structure, e.g., by microfracturing within the structure, which may not be reflected by diffractivc

i measurements of the crystallinity of the material. Such changes in the structural integrity of the material can be measured indirectly by measuring the yield of a product at different levels of structure-modifying treatment. In addition, or alternatively, the change in the molecular structure can include changing the supramolecular structure of the material, oxidation of the material, changing an average molecular weight, changing an average crystallinity, changing a surface area, changing a degree of polymerization, changing a porosity, changing a degree of branching, grafting on other materials, changing a crystalline domain size, or changing an ovcrall domain size.

Unless otherwise defined, ail technical and scientific terms used herein hâve the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or équivalent to those described herein can be used in the practice or testing of the présent invention, suitable methods and materials are described below. Ail publications, patents applications, patents and other référencés mentioned herein are incorporated by reference in their entirety. The materials, methods, and exemples are illustrative only and not intended to be limiting.

Other features and advantages will be apparent from the following detailcd description, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. I is a block diagram illustrating conversion of a feedstock into products and co-products.

FIG. 2 is a block diagram illustrating treatment of the feedstock and the use of the treated feedstock in a fermentation process.

DETAILED DESCRIPTION

Feedstocks that are obtained from plants that hâve been modified with respect to a wild type variety, e.g., by genetic modification or other types of modification, can be proccssed to producc useful intermediates and products such as those described herein. Systems and processes are described herein that can use as feedstock materials e.g., cellulosic and/or lignocellulosic materials that are readily available, but can be difïïcult to process by processes such as fermentation. Many ofthe processes described herein can

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effectively lower the recalcitrance level of the feedstock, making it easier to process, such as by bioprocessing (e.g., with any microorganism described herein, such as a homoacetogen or a heteroacetogcn, and/or any enzyme described herein), thermal processing (e.g., gasification or pyrolysis) or chemîcal methods (e.g., acid hydrolysis or oxidation). The feedstock can be treated or processed using one or more of any of the methods described herein, such as mechanical treatment, chemîcal treatment, radiation, sonication, oxidation, pyrolysis or steam explosion. The various treatment Systems and methods can be used in combinations of two, three, or even four or more of these technologies or others described herein and elsewhere.

In addition to reducing the recalcitrance, the methods outlined above can also sterilize lignocellulosic or cellulosic feedstocks. This can be advantageous because feedstocks can be infected with, for example, a bacteria, a yeast, an insect and/or a fungus, that may hâve a deleterious effect on further processes and/or prematurely dégradé the materials.

Feedstock materials, such as cellulosic and lignocellulosic feedstock materials, can be obtained from plants that hâve been modified with respect to a wild type variety. Such modifications may be for example, by any of the methods described in any patent or patent application rcfcrcnccd herein. As another example, plants may bc modified through the itérative steps of sélection and breeding to obtain desired traits in a plant. Furthermore, the plants can hâve had gcnetic material removed, modified, silenced and/or added with respect to the wild type variety. For example, genetically modified plants can be produced by recombinant DNA methods, where genetic modifications include introducing or modifying spécifie genes from parental varieties, or, for example, by using transgenic breeding wherein a spécifie gene or genes are introduced to a plant from a different species of plant and/or bacteria. Another way to create genetic variation is through mutation breeding wherein new alleles are artifïcially created from endogeneous genes. The artificial genes can bc crcatcd by a variety of ways including treating the plant or seeds with, for example, chemîcal mutagens (e.g., using alkylating agents, epoxides, alkaloids, peroxides, formaldéhyde), irradiation (e.g., X-rays, gamma rays, neutrons, beta particles, alpha particles, protons, deuterons, UV radiation) and température shocking or other external stressing and subséquent sélection techniques. Other methods of providing

modified genes is through errer prone PCR and DNA shuffling followed by insertion of the desired modified DNA into the desired plant or seed, Methods of introducing the desired genetic variation in the seed or plant include, for example, the use of a bacterial carrier, biolistics, calcium phosphate précipitation, electroporation, gene splicing, gene 5 silcncing, lipofection, microinjection and viral carriers.

Feedstock can be derived from a plant including, but not limited to canola, crambe, coconut, maize, mustard, castor bcan, sesame, cottonseed, linsecd, soybean, Arabidopsis phascolus, peanut, alfalfa, wheat, ricc, oat, sorghum, rapeseed, ryc, tritordeum, millet, fescue, rye grass, sugarcane, cranberry, papaya, banana, safïlower, oil 10 palms, flax, muskmelon, apple, cucumbcr, dendrobium, gladiolus, chrysanthemum, liliaccae, cotton, eucalyptus, sunflower, Brassica campestris, Brassica napus, turfgrass, switch grass, cord grass, sugarbeet, coffee, dioscorea, acacia, apricot, artichoke, arugula, asparagus, avocado, barley, beans, beet, blackberry, blueberry, broccoli, brussels sprouts, cabbage, cantaloupe, carrot, cassava, cauliflower, celery, cherry, cilantro, clémentine, 15 com, cotton, Douglas fir, bamboo, seaweed, algac, eggplant, endive, escarole, fennel, fîgs, forest tree, gourd, grape, grapefruit, honcy dew, jîcama, kiwifruit, lettuce, leeks, lemon, lime, loblolly pine, mango, melon, mushroom, nut, oat, okra, onion, orange, parslcy, pea, pcach, pcar, pepper, persimmon, pine, pincapplc, plantain, plum, pomegranate, poplar, potato, oryza sativa, pumpkin, quince, radiata pine, radicchio, radish, raspberry, rye, southem pine, soybean, spinach, squash, strawberry, sweet potato, sweetgum, tangerine, tca, tobacco, tomato, watermelon, wheat, yams, zucchini or mixtures of these. Preferably the feedstock material is derived from plant material not suitable for human consumption such as wood, agricultural waste, grasses such as switchgrass or miscanthus, rice hulls, bagasse, cotton, jute, hemp, flax, bamboo, sisal, 25 abaeâ, straw, com cobs, com stover, hay, coconut hair, seaweed, algae or mixtures of these.

The advantages of plant modification include, for cxamplc, an cnhanccmcnt of résistance to insects, fungal diseases, and other pests and discasc-causing agents; an increased tolérance to herbicides; increased drought résistance; an extended température 30 range; enhanced tolérance to poor soil; enhanced stability or shelf-life; a greater yield; larger fruit size; stronger staiks; enhanced shatter résistance; reduced time to crop maturity; more uniform germination times; higher or modified starch production; cnhanccd nutrient production, such as cnhanced steroid, stcrol, hormone, fatty acid, glycerol, polyhydroxyalkanoate, amino acid, vitamin and/or protein production; modified lignin content; enhanced cellulose, hemiccllulose and/or lignin dégradation; inclusion of a phenotype markcr to allow qualitative détection (e.g., seed coat color); and modified phytatc content. Any feedstock materials derived from these modified plants can also benefit from these many advantages. For examplc, a fccdstock material such as a lignoccllulosic material can have better shelf life, bc casier to process, have a better landto-energy conversion ratio, and/or have a better nutritional value to any microbes that are used in processing of the lignocellulosic material. In addition, any feedstock material derived from such plants can be less expensive and/or more plcntiful. In some cases, modified plants can be grown in a greater variety of climates and/or soil types, for cxample in marginal or deplctcd soils.

Feedstock materials can be obtained from modified plants having an increased résistance to disease. For example, potatoes which have reduced symptoms from the infestation of fungal pathogen Phytophthora infestans arc discussed in U.S. Patent No. 7,122,719. A possible advantage of such résistance is that the yield, quality and shelf life of the fccdstock materials may bc improved.

Feedstock materials can be obtained from modified plants with increased résistance to parasites, for cxample, by cncoding genes for the production of δ-cndotoxins as exemplified in U.S. Patent No. 6,023,013. A possible advantage of such résistance is that the yield, quality and shelf life of the feedstock materials may be improved.

Feedstock materials can be obtained from modified plants having an increased résistance to herbicides. For cxample, the alfalfa plant J-101, as described in U.S. Patent No. 7,566,817, has an increased résistance to glyphosphate herbicides. As a further example, modified plants described in U.S. Patent No. 6,107,549 have an increased résistance to pyridine family herbicides. Furthcrmorc, modified plants described in U.S. Patent No. 7,498,429 have increased résistance to imidazolinoncs. A possible advantage of such résistance is that the yield and quality of the fccdstock materials may be improved.

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Feedstock materials can be obtained from modified plants having an increased stress résistance (for example, water déficit, cold, heat, sait, pest, disease, or nutrient stress). For example, such plants hâve been described in U.S. Patent No. 7,674,952, A possible advantage of such résistance is that the yield and quality of the feedstock materials may be improved. Moreover, such plants may be grown in adverse conditions, e.g., marginal or depleted soil or in a harsh climate.

Feedstock materials can be obtained from modified plants with improved charactcristics such as larger fruits. Such plants have been described in U.S. Patent No. 7,335,812. A possible advantage of such résistance is that the yield and quality of the feedstock materials may be improved.

Feedstock materials can be obtained from modified plants with improved charactcristics such reduced pod shatter. Such plants have been described in U.S. Patent No. 7,659,448. A possible advantage of such résistance is that the yield and quality of the feedstock materials may be improved.

Feedstock materials can be obtained from modified plants having enhaneed or modified starch content. Such plants have been described in U.S. Patent No. 6,538,178. A possible advantage of such modification is that the quality of the feedstock is improved.

Feedstock materials can be obtained from modified plants with a modified oil, fatty acid or glycol production. Such plants have been described in U.S. Patent No. 7,405,344. Fatty acids and oils are excellent substrates for mîcrobial energy-yielding metabolism and may provide an advantage to downstream processing of the feedstock for, for ex ample, fuel production. Fatty acids and oil variation may also be advantageous in changing the viscosity and solubility of various components during downstream processing of the feedstock. The spent feedstock may have a better nutrient mix for use as animal feed or have higher calorie content useful as a direct fuel for buming.

Feedstock materials can be obtained from modified plants with a modified steroid, stcrol and hormone content. Such plants have been described in U.S. Patent No. 6,822,142. A possible advantage is that this may provide a better nutrient mix for microorganisms used in processing of the feedstock. After processing, the spent feedstock may have a better nutrient mix for use as animal feed.

Feedstock materials can be obtained from modified plants with polyhydroxyalkanoate producing ability. Such plants hâve been described in U.S, Patent

No. 6,175,061. Polyhydroxyalkanoates are a useful energy and carbon reserve for various microorganisms and may be bénéficiai to the microorganisms used in downstream feedstock processing. Also, since polyhydroxyalkanoate is biodégradable, it may impart advantages by possibly reducing rccalcitrance in plant material after an aging period of the stored feedstock. Further downstream, the spent feedstock may hâve a better nutrient mix for use as animal feed or hâve higher calorie content useful as a direct fuel for buming.

Feedstock materials can be obtained from modified plants with enhaneed amino acid production. Such plants hâve been described in U.S. Patent No. 7,615,621. A possible advantage is that this may provide a better nutrient mix for microorganisms used in processing of the feedstock. After processing, the spent feedstock may hâve a better nutrient mix for use as animal feed.

Feedstock materials can be obtained from modified plants with elevated synthesis of vitamine. Such plants hâve been described in U.S. Patent No. 6,841,717. A possible advantage is that this may provide a better nutrient mix for microorganisms used in processing of the feedstock. After processing, the spent feedstock may hâve a better nutrient mix for use as animal feed.

Feedstock materials can be obtained from modified plants that dégradé lignin and cellulose in the plant after harvest. Such plants hâve been described in U.S. Patent No. 7,049,485. Feedstock materials can also be obtained from modified plants with modified lignin content. Such plants hâve been described in U.S. Patent No. 7,799,906. A possible advantage of such plants is reduced recalcitrance relative to the wild types of the same plants.

Feedstock materials can be obtained from modified plants with a modified phenotype for easy qualitative détection. Such plants have been described in U.S. Patent No. 7,402,731. A possible advantage is case of managing crops and seeds for different product streams such as biofuels, building materials and animal feed.

Feedstock materials can be obtained from modified plants with a reduced amount of phytate. Such plants have been described in U.S. Patent No, 7,714,187. A possible

A advantage is that this may provide a better nutrient mix for microorganisms used in processing of the feedstock. After processing, the spent feedstock may hâve a better nutrient mix for use as animal feed.

Modified plants and/or plant materiais and methods for making such modifications hâve been described in lhe U.S. Patents and U.S. Published applications listed at the end of this document (immcdiately before the daims), the entire disclosure of each of which is hereby incorporated by reference herein in its entircty.

SYSTEMS FOR TREATING A FEEDSTOCK

FIG. 1 shows one particular process for converting a feedstock, particularly a feedstock obtained at least in part from a modified plant material, into useful intermediates and products. Process 10 includes initially mechamcaily treating the feedstock (12), e.g., to reduce the size of the feedstock 110. The mechanically treated feedstock is then treated with a physical treatment (14) to modify its structure, for example by weakening or microfracturing bonds in the crystalline structure of the material. Next, the structurally modified material may in some cases be subjected to further mechanical treatment (16). This mechanical treatment can bc the same as or different from the initial mechanical treatment. For example, the initial treatment can be a size réduction (e.g., cutting) step followed by a shearing step, while the further treatment can be a grinding or milling step.

The material can then be subjected to further structure-modifying treatment and mechanical treatment, if further structural change (e.g., réduction in recalcitrance) is desired prior to further processing.

Next, the treated material can be processed with a primary processing step 18, e.g., saccharification and/or fermentation, to produce intermediates and products (e.g., energy, fuel, foods and materiais). In some cases, the output of the primary processing step is dircctly useful but, in other cases, requires further processing provided by a post-proccssing step (20). For example, in the case of an alcohol, post-processing may involve distillation and, in some cases, dénaturation.

As described herein, many variations of process 10 can be utilized.

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FIG. 2 shows one particular system that utilizes the steps described above for treating a feedstock and then using the treated feedstock in a fermentation process to produce an alcohol. System 100 includes a module 102 in which a feedstock is initially mechanically treated (step 12, above), a module 104 in which the mechanically treated feedstock is structurally modified (step 14, above), e.g., by irradiation, and a module 106 in which the structurally modified feedstock is subjected to further mechanical treatment (step 16, above). As discussed above, the module 106 may be of the same type as the module 102, or a different type. In some implémentations the structurally modified feedstock can bc retumed to module 102 for further mechanical treatment rather than being further mechanically treated in a separate module 106.

As described herein, many variations of system 100 can be utilized.

After these treatments, which may be repeated as many times as required to obtain desired feedstock properties, the treated feedstock is delivered to a fermentation system 108. Mixing may be performed during fermentation, in which case the mixing is preferably relatively gentle (low shear) so as to minimize damage to shear sensitive ingrédients such as enzymes and other microorganisms. In some embodiments, jet mixing is used, as described in U.S. Serial No. 12/782,694, 13/293,977 and 13/293,985, the complété disclosurcs of which arc incorporated herein by reference.

Referring again to FIG. 2, fermentation produces a crude éthanol mixture, which flows into a holding tank 110. Water or other solvent, and other non-ethanol components, arc stripped from the crude éthanol mixture using a stripping column 112, and the éthanol is then distilled using a distillation unit 114, e.g., a rectifier. Distillation may be by vacuum distillation. Finally, the éthanol can be dried using a molecular sieve 116 and/or denatured, if necessary, and output to a desired shipping method.

In some cases, the Systems described herein, or components thereof, may be portable, so that the System can bc transported (e.g., by rail, truck, or marine vessel) from one location to another. The method steps described herein can bc performed at onc or more locations, and in some cases onc or more of the steps can bc performed in transit. Such mobile processing is described in U.S. Serial No. 12/374,549 and International Application No. WO 2008/011598, the fiill disclosurcs of which arc incorporated herein by reference.

Any or ail of the method steps described herein can be performed at ambient température. If desired, cooling and/or heating may be employed during certain steps. For example, the feedstock may be cooled during mechanical treatment to increase its brittlcness. In some embodiments, cooling is employed before, during or after the initial mechanical treatment and/or the subséquent mechanical treatment. Cooling may be performed as described in U.S. Serial No. 12/502,629, now U.S. Patent No. 7,900,857 the iull disclosure of which is incorporatcd herein by reference. Morcover, the température in the fermentation system 108 may be controlled to enhance saccharification and/or fermentation.

The individual steps of the methods described above, as well as the materials used, will now be described in further detail.

PHYSICAL TREATMENT

Physical treatment processes can include one or more of any of those described herein, such as mechanical treatment, chemical treatment, irradiation, sonication, oxidation, pyrolysis or steam explosion. Treatment methods can be used in combinations of two, three, four, or even ail of these technologies (in any order), When more than one treatment method is used, the methods can be applied at the same time or at different fîmes. Other processes that change a molecular structure of a feedstock may also be used, alone or in combination with the proccsscs disclosed herein.

Mechanical Treatments

In some cases, methods can include mechanically treating the feedstock. Mechanical treatments include, for example, cutting, milling, pressing, grinding, shearing and chopping. Milling may include, for example, bail milling, hammer milling, rotor/stator dry or wet milling, freezer milling, blade milling, knife milling, disk milling, relier milling or other types of milling. Other mechanical treatments include, e.g., stonc grinding, cracking, mechanical ripping or tearing, pin grinding or air attrition milling.

Mechanical treatment can be advantageous for opening up,” stressing,” breaking and shattering cellulosic or lignocellulosic materials in the feedstock, making the cellulose of the materials more susceptible to chain scission and/or réduction of

crystallinity. The open materials can also be more susceptible to oxidation when irradiatcd.

In some cases, the mechanical treatment may include an initial préparation of the feedstock as received, e.g., size réduction of materials, such as by cutting, grinding, shearing, pulverizing or chopping. For example, in some cases, loose feedstock (e.g., recycled paper, starchy materials, or switchgrass) is prepared by shearing or shredding.

Alternatively, or in addition, the feedstock material can first be physically treated by onc or more of the other physical treatment methods, e.g., chemical treatment, radiation, sonication, oxidation, pyrolysis or steam explosion, and then mechanically treated. This sequence can be advantageous since materials treated by one or more of the other treatments, e.g., irradiation or pyrolysis, tend to be more brittle and, therefore, it may be easier to further change the molecular structure of the material by mechanical treatment.

In some embodiments, the feedstock is in the form of a fibrous material, and mechanical treatment includes shearing to expose fîbers of the fibrous material. Shearing can be performed, for example, using a rotary knife cutter. Other methods of mechanically treating the feedstock include, for exemple, milling or grinding. Milling may bc performed using, for example, a hammer mill, bail mill, colloid mill, conical or cône mill, disk mill, edge mill, Wiley mill or grist mill. Grinding may be performed using, for example, a stonc grinder, pin grinder, coffee grinder, or burr grinder. Grinding may be provided, for example, by a reciprocating pin or other element, as is the case in a pin mill. Other mechanical treatment methods include mechanical ripping or tearing, other methods that apply pressure to the material, and air attrition milling. Suitable mechanical treatments further include any other technique that changes the molecular structure of the feedstock.

If desired, the mechanically treated material can be passed through a screen, e.g., having an average opening size of 1.59 mm or less (1/16 inch, 0.0625 inch). In some embodiments, shearing, or other mechanical treatment, and screening arc performed concurrently. For example, a rotary knife cutter can be used to concurrently shear and screen the feedstock. The feedstock is sheared between stationary bladcs and rotating bladcs to provide a sheared material that passes through a screen, and is captured in a bin.

The feedstock can be mechanically treated in a dry state (e.g., having little or no free water on its surface), a hydrated state (e.g,, having up to ten percent by weight absorbed water), or in a wet state, e.g., having between about 10 percent and about 75 percent by weight water. The fiber source can even be mechanically treated while partially or fully submerged under a liquid, such as water, éthanol or isopropanol.

The feedstock can also be mechanically treated under a gas (such as a stream or atmosphère of gas other than air), e.g., oxygen or nitrogen, or steam.

If desired, lignin can bc removed from any of the fibrous materials that include lignin. Also, to aid in the breakdown of the materials that include cellulose, the material can be treated prior to or during mechanical treatment or irradiation with heat, a chemical (e.g., minerai acid, base or a strong oxidizer such as sodium hypochlorite) and/or an enzyme. For example, grinding can be performed in the presence of an acid.

Mechanical treatment système can be confîgured to produce streams with spécifie morphology characteristics such as, for example, surface area, porosity, bulk density, and, in the case of fibrous feedstocks, fiber characteristics such as length-to-width ratio.

In some embodiments, a BET surface area of the mechanically treated material is greater than 0.1 m²/g, e.g., greater than 0.25 m^z/g, greater than 0.5 m²/g, greater than 1.0 m²/g, greater than 1.5 m²/g, greater than 1.75 m²/g, greater than 5.0 m²/g, greater than 10 m²/g, greater than 25 m²/g, greater than 35 m²/g, greater than 50m²/g, greater than 60 m²/g, greater than 75 m²/g, greater than 100 m²/g, greater than 150 m²/g, greater than 200 m²/g, or even greater than 250 m²/g.

A porosity of the mechanically treated material can be, e.g., greater than 20 percent, greater than 25 percent, greater than 35 percent, greater than 50 percent, greater than 60 percent, greater than 70 percent, greater than 80 percent, greater than 85 percent, greater than 90 percent, greater than 92 percent, greater than 94 percent, greater than 95 percent, greater than 97.5 percent, greater than 99 percent, or even greater than 99.5 percent.

In some embodiments, after mechanical treatment the material has a bulk density of less than 0.75 g/cm³, e.g., less than about 0.7, 0.65, 0.60, 0.50, 0.35, 0.25, 0.20,0.15, 0.10, 0.05, or less, e.g., less than 0.025 g/cm³. Bulk density is determined using ASTM D1895B. Briefly, the method involves filling a measuring cylinder of known volume

with a sample and obtaining a weight of the sample. The bulk density is calculated by dividing the weight of the sample in grams by the known volume of the cylinder in cubic centimeters.

If the feedstock is a fibrous material the fibers of the mechanically treated material can have a relatively large average lcngth-to-diameter ratio (e.g., greater than 20-to»l), even if they have been shearcd more than once. In addition, the fibers of the fibrous materials described herein may have a relatively narrow length and/or length-todiameter ratio distribution.

As used herein, average fiber widths (e.g,, diameters) are those determined optically by randomly selecting approximately 5,000 fibers. Average fiber lengths are corrected length-weighted lengths. BET (Brunaucr, Emmct and Tellcr) surface areas are multi-point surface areas, and porosities arc those determined by mercury porosimetry.

If the feedstock is a fibrous material the average length-to-diameter ratio of fibers of the mechanically treated material can be, e.g., greater than 8/1, e.g., greater than 10/1, greater than 15/1, greater than 20/1, greater than 25/1, or greater than 50/1. An average fiber length of the mechanically treated material can be, e.g., between about 0.5 mm and

2.5 mm, e.g., between about 0.75 mm and 1.0 mm, and an average width (e.g., diameter) of the second fibrous material 14 can be, e.g., between about 5 gm and 50 gm, e.g., between about 10 gm and 30 gm.

In some embodiments, if the feedstock is a fibrous material the standard déviation of the fiber length of the mechanically treated material can be less than 60 percent of an average fiber length of the mechanically treated material, e.g., less than 50 percent of the average length, less than 40 percent of the average length, less than 25 percent of the average length, less than 10 percent of the average length, less than 5 percent of the average length, or even less than 1 percent of the average length.

In some situations, it can be désirable to prépare a low bulk density material, densify the material (e.g., to makc it casier and less costly to transport to another site), and then revert the material to a lower bulk density state. Dcnsificd materials can be processcd by any of the methods described herein, or any material proccssed by any of the methods described herein can be subsequcntly dcnsificd, e.g., as disclosed in U.S.

Serial No. 12/429,045 now U.S. Patent No. 7,932,065 and WO 2008/073186, the full disclosures of which are incorporated herein by reference.

Radiation Treatment

One or more radiation processing sequences can be used to process the feedstock, and to provide a structurally modified material which functions as input to further processing steps and/or sequences. Irradiation can, for example, rcduce the molecular weight and/or crystallinity of feedstock. Radiation can also sterilize the materials, or any media needed to bioprocess the material.

In some embodiments, energy deposited in a material that releases an électron from its atomic orbital is used to irradiaie the materials. The radiation may be provided by (1) heavy charged particles, such as alpha particles or protons, (2) électrons, produced, for example, in beta decay or électron beam accelerators, or (3) electromagnetic radiation, for example, gamma rays, x rays, or ultraviolet rays. In one approach, radiation produced by radioactive substances can be used to irradiate the feedstock. In another approach, electromagnetic radiation (e.g., produced using électron beam emitters) can be used to irradiate the feedstock. In some embodiments, any combination in any order or concurrcntly of (1) through (3) may be utilized. The doses applied dépend on the desired effect and the particular feedstock.

In some instances when chain scission is désirable and/or polymer chain functionalization is désirable, particles heavicr than électrons, such as protons, hélium nuclei, argon ions, silicon ions, néon ions, carbon ions, phosphorus ions, oxygen ions or nitrogen ions can be utilized. When ring-opening chain scission is desired, positively charged particles can be utilized for their Lewis acid properties for enhanced ringopening chain scission. For example, when maximum oxidation is desired, oxygen ions can be utilized, and when maximum nitration is desired, nitrogen ions can be utilized. The use of heavy particles and positively charged particles is described in U.S. Serial No. 12/417,699, now U.S. Patent No. 7,931,784, the full disclosure of which is incorporated herein by reference.

In one method, a first material that is or includes cellulose having a first number average molecular weight (Mni) is irradiated, e.g., by treatment with ionizing radiation

(e.g., in the form of gamma radiation, X-ray radiation, 100 nm to 280 nm ultraviolet (UV) light, a beam of électrons or other charged particles) to provide a second material that includes cellulose having a second number average molecular weight (M_N2) lower than the first number average molecular weight. The second material (or the first and second material) can bc combined with a mîcroorganism (with or without enzyme treatment) that can utilize the second and/or first material or its constituent sugars or lignin to produce an intermediate or product, such as those described herein.

Since the second matériel includes cellulose having a reduced molecular weight relative to the first material, and in some instances, a reduced crystallinity as well, the second material is generally more dispersible, swellable and/or soluble, e.g., in a solution containing a mîcroorganism and/or an enzyme. These properties make the second material casier to process and more susceptible to chemical, enzymatic and/or biological attack relative to the first material, which can greatly improve the production rate and/or production level of a desired product, e.g., éthanol.

In some embodiments, the second number average molecular weight (Mn₂) is lower than the first number average molecular weight (Mnj) by more than about 10 percent, e.g., more than about 15, 20, 25, 30, 35, 40, 50 percent, 60 percent, or even more than about 75 percent.

ln some instances, the second material includes cellulose that has a crystallinity (C₂) that is lower than the crystallinity (Ci) of the cellulose of the first material. For cxample, (C₂) can be lower than (Ci) by more than about 10 percent, e.g., more than about 15, 20, 25, 30, 35,40, or even more than about 50 percent.

In some embodiments, the starting crystallinity index (prior to irradiation) is from about 40 to about 87.5 percent, e.g., from about 50 to about 75 percent or from about 60 to about 70 percent, and the crystallinity index after irradiation is from about 10 to about 50 percent, e.g., from about 15 to about 45 percent or from about 20 to about 40 percent. However, in some embodiments, e.g., after extensive irradiation, it is possible to have a crystallinity index of lower than 5 percent. In some embodiments, the material after irradiation is substantially amorphous.

In some embodiments, the starting number average molecular weight (prior to irradiation) is from about 200,000 to about 3,200,000, e.g., from about 250,000 to about

.V

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, e.g., after extensive irradiation, it is possible to hâve a number average molecular weight 5 of less than about 10,000 or even less than about 5,000.

In some embodiments, the second material can hâve a level of oxidation (O₂) that is higher than the level of oxidation (Oi) of the first material. A higher level of oxidation of the material can aid in its dispcrsabilîty, swcllability and/or solubility, further enhancing the material’s susccptibility to chemîcal, enzymatic or biological attack. In some embodiments, to increase the level of the oxidation of the second material relative to the first material, the irradiation is performed under an oxidizing environment, e.g., under a blankct of air or oxygen, producîng a second material that is more oxidized than the first material. For example, the second material can hâve more hydroxyl groups, aldéhyde groups, ketone groups, ester groups or carboxylic acid groups, which can increase its hydrophilicity.

Ionizine Radiation

Each form of radiation ionizes the carbon-containing material via particular interactions, as determined by the energy of the radiation. Heavy charged particles primarily ionize matter via Coulomb scattering; furthermore, these interactions produce 20 energetic électrons that may further ionize matter. Alpha particles are identical to the nucléus of a hélium atom and are produced by the alpha decay of various radioactive nucleî, such as isotopes of bismuth, polonium, astatine, radon, francium, radium, several actinides, such as actinium, thorium, uranium, neptunium, curium, californium, américium, and plutonium.

When particles arc utilized, they can bc ncutral (uneharged), positively charged or ncgativcly charged. When charged, the charged particles can bcar a single positive or négative charge, or multiple charges, e.g., one, two, three or even four or more charges. In instances in which chain scission is desired, positively charged particles may be désirable, in part due to their acidic nature. When particles are utilized, the particles can hâve the mass of a resting électron, or greater, e.g., 500,1000, 1500, 2000, 10,000 or even 100,000 times the mass of a resting électron. For example, the particles can hâve a mass of from about 1 atomic unit to about 150 atomic units, e.g., from about 1 atomic unit to about 50 atomic units, or from about 1 to about 25, e.g., 1, 2, 3,4, 5, 10, 12 or 15 amu. Accéléra tors used to accelerate the particles can be electrostatic DC, electrodynamic DC, RF linear, magnetic induction linear or continuons wave. For example, cyclotron type accelerators are available from IBA, Belgium, such as the Rhodotron® systern, while DC type accelerators arc available from RDI, now IBA Industrial, such as the Dynamitron®. Ions and ion accelerators are discussed in Introductory Nuclcar Physics, Kenneth S, Krane, John Wiley & Sons, Inc. (1988), Krsto Prclcc, F1Z1KA B 6 (1997) 4, 177-206, Chu, William T., “Ovcrvicw of Light-lon Bcam Therapy” Columbus-Ohio, 1CRU-IAEA Meeting, 18-20 March 2006, Iwata, Y. et al., “Altemating-Phase-Focuscd IH-DTL for Hcavy-Ion Medical Accelerators” Procecdings of EPAC 2006, Edinburgh, Scotland and Leaner, C.M. et al, “Status of the Superconducting ECR Ion Source Venus” Procecdings of EPAC 2000, Vienna, Austria.

Gamma radiation has the advantage of a significant pénétration depth into a variety of materials. Sources of gamma rays include radioactive nuclei, such as isotopes of cobalt, calcium, technicium, chromium, gallium, indium, iodine, iron, krypton, samarium, sélénium, sodium, thalium, and xénon.

Sources of x rays include électron beam collision with métal targets, such as tungsten or molybdcnum or alloys, or compact light sources, such as those produced commercially by Lyncean.

Sources for ultraviolet radiation include deuterium or cadmium lamps.

Sources for infrared radiation include sapphire, zinc, or selenide window ceramic lamps.

Sources for microwaves include klystrons, Slevin type RF sources, or atom bcam sources that employ hydrogen, oxygen, or nitrogen gases.

In some embodiments, a beam of électrons is used as the radiation source. A beam of électrons has the advantages of high dose rates (e.g., 1, 5, or even 10 Mrad per second), high throughput, less containment, and less confinement equipment. Electrons can also be more efficient at causing chain scission. In addition, électrons having

( energies of 4-10 MeV can hâve a pénétration depth of 5 to 30 mm or more, such as 40 mm.

Electron beams can be generatcd, e.g,, by electrostatic generators, cascade generators, transformer generators, low energy accelerators with a scanning system, low energy accelerators with a linear cathode, linear accelerators, and pulsed accelerators. Electrons as an ionizing radiation source can be useful, e.g., for relatively thin sections of material, e.g., less than 0.5 înch, e.g., less than 0.4 inch, 0.3 inch, 0.2 inch, or less than 0.1 inch. In some embodiments, the energy of each électron of the électron bcam is from about 0.3 MeV to about 2.0 MeV (million électron volts), e.g., from about 0.5 MeV to about 1.5 MeV, or from about 0.7 MeV to about 1.25 MeV.

Electron beam irradiation devices may be procured commercially from Ion Bcam Applications, Louvain-la-Ncuve, Belgium or the Titan Corporation, San Diego, CA. Typical électron energies can be 1 MeV, 2 MeV, 4.5 MeV, 7.5 MeV, or 10 MeV. Typical électron beam irradiation device power can be 1 kW, 5 kW, 10 kW, 20 kW, 50 kW, 100 kW, 250 kW, or 500 kW. The level of depolymerization of the feedstock dépends on the électron energy used and the dose applied, while exposure time dépends on the power and dose. Typical doses may take values of 1 kGy, 5 kGy, 10 kGy, 20 kGy, 50 kGy, 100 kGy, or 200 kGy. In a some embodiments énergies between 0.25-10 MeV (e.g., 0.5-0.8 MeV, 0.5-5 MeV, 0.8-4 MeV, 0.8-3 MeV, 0.8-2 MeV or 0.8-1.5 MeV) can be used. In some embodiments doses between 1-100 Mrad (e.g., 2-80 Mrad, 5-50 Mrad,

5-40 Mrad, 5-30 Mrad or 5-20 Mrad) can be used. In some preferred embodiments, an energy between 0.8-3 MeV (e.g., 0.8-2 MeV or 0.8-1.5 MeV) combined with doses between 5-50 Mrad (e.g., 5-40 Mrad, 5-30 Mrad or 5-20 Mrad) can be used.

Ion Particle Beams

Particles heavicr than électrons can be utîlized to irradiate materials, such as carbohydrates or materials that include carbohydrates, e.g., ccllulosic materials, lignocellulosic materials, starchy materials, or mixtures of any of these and others described herein. For example, protons, hélium nuclei, argon ions, silicon ions, néon ions carbon ions, phosphorus ions, oxygen ions or nitrogen ions can be utilized. In some embodiments, particles heavicr than électrons can induce higher amounts of chain

scission (relative to lighter particles). In some instances, positively charged particles can induce higher amounts of chain scission than negatively charged particles due to their acidity.

Heavîer particle beams can be generated, e.g., using linear accelerators or cyclotrons. In some embodiments, the energy of each particle of the beam is from about l .0 MeV/atomic unit (MeV/amu) to about 6,000 MeV/atomic unit, e.g., from about 3 MeV/ atomic unit to about 4,800 MeV/atomic unit, or from about 10 MeV/atomic unit to about 1,000 MeV/atomic unit.

In certain embodiments, ion beams used to irradiate carbon-containing matériels, e.g., materiais obtained from plants, can include more than one type of ion. For example, ion beams can include mixtures of two or more (e.g., three, four or more) different types of ions. Exemplary mixtures can include carbon ions and protons, carbon ions and oxygen ions, nitrogen ions and protons, and iron ions and protons. More generally, mixtures of any of the ions discussed above (or any other ions) can be used to form irradlating ion beams. In particular, mixtures of relatively light and relatively heavier ions can be used in a single ion beam.

In some embodiments, ion beams for irradiating materiais include positivelychargcd ions. The positively charged ions can include, for exemple, positively charged hydrogen ions (e.g., protons), noble gas ions (e.g., hélium, néon, argon), carbon ions, nitrogen ions, oxygen ions, silicon atoms, phosphorus ions, and métal ions such as sodium ions, calcium ions, and/or iron ions. Without wishing to be bound by any theory, it is belicved that such positively-charged ions behave chemically as Lewis acid moieties when exposed to materiais, initiating and sustaining cationic ring-opening chain scission reactions in an oxidative environment.

In certain embodiments, ion beams for irradiating materiais include negativelycharged ions. Negatively charged ions can include, for example, negatively charged hydrogen ions (e.g., hydridc ions), and negatively charged ions of various relatively clcctroncgativc nuclci (e.g., oxygen ions, nitrogen ions, carbon ions, silicon ions, and phosphorus ions). Without wishing to be bound by any theory, it is believed that such negatively-charged ions behave chemically as Lewis base moieties when exposed to

materials, causing anionie ring-opening chain scission reactions in a reducing environment.

In some embodiments, beams for irradiating materials can include neutral atoms. For example, any one or more of hydrogen atoms, hélium atoms, carbon atoms, nitrogen atoms, oxygen atoms, néon atoms, silicon atoms, phosphorus atoms, argon atoms, and iron atoms can be included in beams that are used for irradiation. In general, mixtures of any two or more of the above types of atoms (e.g., three or more, four or more, or even more) can bc présent in the beams.

In certain embodiments, ion beams used to irradiate materials include singlycharged ions such as one or more of H⁺, H, He⁺, Ne*, Ar*, C*, C', O*, O', N*, N’, Si*, Si', P⁺, P’, Na⁺, Ca⁺, and Fe*. In some embodiments, ion beams can include multiply-charged ions such as onc or more of C²*, C³*, C⁴⁺, N³*, N⁵*, N³, Ο²*, Ο²’, Oj²’, Si²*, Si⁴*, Si²', and Si⁴'. In general, the ion beams can also include more complex polynuclear ions that bear multiple positive or négative charges. In certain embodiments, by virtue of the structure of the polynuclear ion, the positive or négative charges can be effectively distributed over substantially the entire structure of the ions. In some embodiments, the positive or négative charges can be somewhat localizcd over portions of the structure of the ions.

Electromagnetic Radiation

In embodiments in which the irradiating is performed with electromagnetic radiation, the electromagnetic radiation can hâve, e.g., energy per photon (in électron volts) of greater than lO² eV, e.g., greater than ΙΟ³, ΙΟ⁴,10⁵, ÎO⁶, or even greater than ÎO⁷ eV. In some embodiments, the electromagnetic radiation has energy per photon of between ÎO⁴ and 10⁷, e.g., between 10⁵ and 10^û eV, The electromagnetic radiation can hâve a frequency of, e.g., greater than 10¹⁶ hz, greater than 10¹⁷ hz, ΙΟ¹⁸, 10¹⁹, 10²⁰, or even greater than 10²¹ hz. Typical doses may takc values of greater than 1 Mrad (e.g., greater than 1 Mrad, greater than 2 Mrad). In some embodiments, the electromagnetic radiation has a frequency of between 10¹⁸ and 10²² hz, e.g., between 10¹⁹ to 10²¹ hz. In some embodiment doses between 1-100 Mrad (e.g., 2-80 Mrad, 5-50 Mrad, 5-40 Mrad,

5-30 Mrad or 5-20 Mrad) can be used.

t

I

Quenching and Controlled Functionalization

After treatment with ionizing radiation, any of the materials or mixtures described herein may become ionized; that is, the treated material may include radicals at levels that are détectable with an électron spin résonance spectrometer. If an ionized feedstock rcmains in the atmosphère, it will be oxidized, such as to an extent that carboxylic acid groups are generated by reacting with the atmospheric oxygen. In some instances with some materials, such oxidation is desired becausc it can aid in the further breakdown in molecular weight of the carbohydratc-containing biomass, and the oxidation groups, e.g., carboxylic acid groups can bc hclpful for solubility and microorganism utilization in some instances. Howcver, since the radicals can “live” for some time after irradiation, e.g., longer than 1 day, 5 days, 30 days, 3 months, 6 months or even longer than 1 year, material properties can continue to change over time, which in some instances, can be undesirable. Thus, it may be désirable to quench the ionized material.

After ionization, any ionized material can be quenched to reduce the level of radicals in the ionized material, e.g., such that the radicals are no longer détectable with the électron spin résonance spectrometer. For example, the radicals can be quenched by the application of a sufficient pressure to the material and/or by utilizing a fluid in contact with the ionized material, such as a gas or liquid, that rcacts with (qucnchcs) the radicals. Using a gas or liquid to at least aid in the quenching of the radicals can be used to functîonalizc the ionized material with a desired amount and kind of functîonal groups, such as carboxylic acid groups, enol groups, aldéhyde groups, nitro groups, nitrile groups, amino groups, alkyl amino groups, alkyl groups, chloroalkyl groups or chlorofluoroalkyl groups.

In some instances, such quenching can improve the stability of some of the ionized materials. For example, quenching can improve the résistance of the material to oxidation. Functionalization by quenching can also improve the solubility of any material described herein, can improve its thermal stability, and can improve material utilization by various microorganisms. For cxamplc, the functîonal groups imparted to the material by the quenching can act as receptor sites for attachment by microorganisms, e.g., to enhance cellulose hydrolysis by various microorganisms.

t

In some embodiments, quenching includes an application of pressure to the ionized material, such as by mcchanically deforming the material, e.g., directly mechanically compressing the material in one, two, or three dimensions, or applying pressure to a fluid in which the material is immersed, e.g., isostatic pressing. In such instances, the deformation of the malerial itself brings radicale, which are often trapped in crystalline domains, in close enough proximity so that the radicale can recombine, or react with another group. In some instances, the pressure is applied together with the application of heat, such as a sufficicnt quantity of heat to elevate the température of the material to above a melting point or softening point of a component of the material, such as lignin, cellulose or hemicellulose. Heat can improve molecular mobility in the material, which can aid in the quenching of the radicals. When pressure is utilized to quench, the pressure can be greater than about 1000 psi, such as greater than about 1250 psi, 1450 psi, 3625 psi, 5075 psi, 7250 psi, 10000 psi or even greater than 15000 psi.

In some embodiments, quenching includes contacting the ionized material with a fluid, such as a liquid or gas, e.g., a gas capable of reacting with the radicals, such as acetylene or a mixture of acetylene in nitrogen, ethylene, chlorinated ethylenes or chlorofluorocthylcnes, propylenc or mixtures of these gases. In other particular embodiments, quenching includes contacting the ionized material with a liquid, e.g., a liquid soluble in, or at least capable of penetrating into the material and reacting with the radicals, such as a diene, such as 1,5-cyclooctadiene. In some spécifie embodiments, quenching includes contacting the material with an antioxidant, such as Vitamin E. If desired, the feedstock can include an antioxidant dispersed thercin, and the quenching can corne from contacting the antioxidant dispersed in the feedstock with the radicals.

Functionalization can be enhanced by utilizing heavy charged ions, such as any of the heavier ions described herein. For example, if it is desired to cnhance oxidation, charged oxygen ions can be utilized for the irradiation. If nitrogen functional groups are desired, nitrogen ions or anions that include nitrogen can bc utilized. Likcwisc, if sulfur or phosphorus groups arc desired, sulfur or phosphorus ions can bc used in the irradiation.

I

Doses

In some instances, the irradiation is performed at a dosage rate of greater than about 0.25 Mrad per second, e.g., greater than about 0.5,0,75,1.0,1.5, 2.0, or even greater than about 2.5 Mrad per second. In some embodiments, the irradiating is performed at a dose rate of between 5.0 and 1500.0 kilorads/hour, e.g., between 10.0 and 750.0 kilorads/hour or between 50.0 and 350.0 kilorads/hour. In some embodiments, irradiation is performed at a dose rate of greater than about 0.25 Mrad per second, e.g., greater than about 0.5,0.75, 1,1.5, 2, 5, 7, 10,12,15, or even greater than about 20 Mrad per second, e.g., about 0.25 to 2 Mrad per second.

In some embodiments, the irradiating (with any radiation source or a combination of sources) is performed until the material receives a dose of 0.25 Mrad, e.g., at least 1.0, 2.5, 5.0, 8.0, 10, 15, 20, 25, 30, 35, 40, 50, or even at least 100 Mrad. In some embodiments, the irradiating is performed until the material receives a dose of between 1.0 Mrad and 6.0 Mrad, e.g., between 1.5 Mrad and 4.0 Mrad, 2 Mrad and 10 Mrad, 5 Mrad and 20 Mrad, 10 Mrad and 30 Mrad, 10 Mrad and 40 Mrad, or 20 Mrad and 50 Mrad. In some embodiments, the irradiating is performed until the material receives a dose of from about 0.1 Mrad to about 500 Mrad, from about 0.5 Mrad to about 200 Mrad, from about 1 Mrad to about 100 Mrad, or from about 5 Mrad to about 60 Mrad. In some embodiments, a relatively low dose of radiation is applied, e.g., less than 60 Mrad.

Sonicatlon

Sonication can reduce the molecular weight and/or crystallinity of materials, such as one or more of any of the materials described herein, e.g., one or more carbohydrate sources, such as cellulosic or lignocellulosic materials, or starchy materials. Sonication can also be used to sterilize the materials. As discussed above with regard to radiation, the process parameters used for sonication can be varied depending on various factors, e.g., depending on the lignin content of the feedstock. For exemple, feedstocks with higher lignin levels generally require a higher résidence time and/or energy level, resulting in a higher total energy delivered to the feedstock.

In one method, a first material that includes cellulose having a first number average molecular weight (Mm) is dispersed in a medium, such as water, and sonicated

and/or otherwise cavitated, to provide a second material that includes cellulose having a second number average molecular weight (Mnï) lower than the first number average molecular weight. The second material (or the first and second material in certain embodiments) can be combined with a microorganism (with or without enzyme treatment) that can utîlize the second and/or first material to produce an intermediate or product.

Sînce the second material includes cellulose having a reduced molecular weight relative to the first material, and in some instances, a rcduccd crystallinity as well, the second material is generally more dispersible, swellable, and/or soluble, e.g., in a solution containing a microorganism.

In some embodiments, the second number average molecular weight (Mn₂) is lower than the first number average molecular weight (Μχι) by more than about 10 percent, e.g., more than about 15,20, 25,30, 35, 40, 50 percent, 60 percent, or even more than about 75 percent.

In some instances, the second material includes cellulose that has a crystallinity (C₂) that is lower than the crystallinity (Cj) of the cellulose of the first material. For cxample, (C₂) can be lower than (Ci) by more than about 10 percent, e.g., more than about 15, 20,25, 30, 35, 40, or even more than about 50 percent.

In some embodiments, the starting crystallinity index (prior to sonication) is from about 40 to about 87.5 percent, e.g., from about 50 to about 75 percent or from about 60 to about 70 percent, and the crystallinity index after sonication is from about 10 to about 50 percent, e.g., from about 15 to about 45 percent or from about 20 to about 40 percent. However, in certain embodiments, e.g., after extensive sonication, it is possible to hâve a crystallinity index of lower than 5 percent. In some embodiments, the material after sonication is substantially amorphous.

In some embodiments, the starting number average molecular weight (prior to sonication) is from about 200,000 to about 3,200,000, e.g., 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 sonication 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,

, ¹

e.g., after extensive sonication, it is possible to have a number average molecular weight of less than about 10,000 or even less than about 5,000.

In some embodiments, the second material can have a level of oxidation (O₂) that is higher than the level of oxidation (Oj) of the first material. A higher level of oxidation of the material can aid in its dispersabilîty, swellability and/or solubility, further cnhancing the material’s susceptibility to chemical, cnzymatic or mîcrobial attack. In some embodiments, to increase the level of the oxidation of the second material relative to the first material, the sonication is performed in an oxidizing medium, producing a second material that is more oxidized than the first material. For example, the second material can have more hydroxyl groups, aldéhyde 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 can include an oxidant, such as a peroxide (e.g., hydrogen peroxide), a dispersing agent and/or a buffer. Examples of dispersing agents include ionic dispersing agents, e.g., sodium lauryl sulfate, and non-ionic dispersing agents, e.g., poly(ethylene glycol).

In other embodiments, the sonication medium is non-aqucous. For example, the sonication can be performed in a hydrocarbon, e.g., toluene or heptane, an ether, c.g„ diethyl ether or tetrahydrofuran, or even in a liquefied gas such as argon, xénon, or nitrogen.

Pvrolysls

One or more pyrolysis processing sequences can be used to process carboncontaining materials from a wide variety of different sources to extract useful substances from the materials, and to provide partially degraded materials which fonction as input to forther processing steps and/or sequences. Pyrolysis can also be used to sterilize the materials. Pyrolysis conditions can bc varied depending on the charactcristics of the feedstock and/or other factors. For example, fccdstocks with higher lignin levels may requirc a higher température, longer résidence time, and/or introduction of higher levels of oxygen during pyrolysis.

ln one example, a first material that includes cellulose having a first number average molecular weight (Mni) is pyrolyzed, e.g., by heating the first material in a tube fumace (in the presence or absence of oxygen), to provide a second material that includes cellulose having a second number average molecular weight (M_N2) lower than the first 5 number average molecular weight.

Since the second material includes cellulose having a reduced molecular weight relative to the first material, and in some instances, a reduced ciystallinity as well, the second material is generally more dispcrsiblc, swellable and/or soluble, e.g., in a solution containing a microorganism.

In some embodiments, the second number average molecular weight (Mnî) is lower than the first number average molecular weight (Μχι) by more than about 10 percent, e.g., more than about 15, 20,25, 30, 35,40, 50 percent, 60 percent, or even more than about 75 percent.

In some instances, the second material includes cellulose that has a crystallinity (C₂) that is lower than the crystallinity (Ci) of the cellulose of the first material. For example, (C₂) can be lower than (Ci) by more than about 10 percent, e.g., more than about 15, 20, 25, 30, 35,40, or even more than about 50 percent.

In some embodiments, the starting crystallinity (prior to pyrolysis) is from about 40 to about 87.5 percent, e.g., from about 50 to about 75 percent or from about 60 to about 70 percent, and the crystallinity index after pyrolysis is from about 10 to about 50 percent, e.g., from about 15 to about 45 percent or from about 20 to about 40 percent. However, in certain embodiments, e.g., after extensive pyrolysis, it is possible to have a crystallinity index of lower than 5 percent. In some embodiments, the material after pyrolysis is substantially amorphous.

In some embodiments, the starting number average molecular weight (prior to pyrolysis) îs from about 200,000 to about 3,200,000, e.g., 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., fforn about 60,000 to about 150,000 or from about 70,000 to about 125,000. However, in some embodiments,

e.g., after extensive pyrolysis, it is possible to have a number average molecular weight of less than about 10,000 or even less than about 5,000.

In some embodiments, the second material can have a level of oxidation (O₂) that is higher than the level of oxidation (Oj) of the first material. A higher level of oxidation of the material can aid in its dispersability, swellability and/or solubility, further enhancing the susceptibility of the material to chemical, enzymatic or microbial attack,

In some embodiments, to increase the level of the oxidation of (he second material relative to the first material, the pyrolysis is performed in an oxidizing environment, producing a second material that is more oxidized than the first material. For example, the second material can have more hydroxyl groups, aldéhyde groups, kctonc groups, ester groups or carboxylic acid groups, than the first material, thereby increasing the o hydrophi licity of the materi al.

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

Oxidation

One or more oxidative processing sequences can be used to process carboncontaining materials from a wide variety of different sources to extract useful substances from the materials, and to providc partially degraded and/or altcrcd material which functions as input to further processing steps and/or sequences. The oxidation conditions 20 can be varied, e.g., depending on the lignin content of the feedstock, with a higher degree of oxidation generally being desired for higher lignin content feedstocks.

In one method, a first material that includes cellulose having a first number average molecular weight (Mni) and having a first oxygen content (O|) is oxidized, e.g., by heating the first material in a stream of air or oxygen-cnriched air, to provide a second 25 material that includes cellulose having a second number average molecular weight (Μχ₂) and having a second oxygen content (O₂) higher than the first oxygen content (Oj).

The second number average molecular weight of the second material is generally lower than the first number average molecular weight of the first material. For examplc, the molecular weight may be reduced to the same extent as discussed above with respect 30 to the other physicaï treatments. The crystallinity of the second material may also be

reduced to the same extent as discussed above with respect to the other physical treatmcnts.

In some embodiments, the second oxygen content is at least about five percent higher than the first oxygen content, e.g., 7.5 percent higher, 10.0 percent higher, 12.5 percent higher, 15.0 percent higher or 17.5 percent higher. In some preferred embodiments, the second oxygen content is at least about 20.0 percent higher than the first oxygen content of the first material. Oxygen content is measured by elemental analysis by pyrolyzing a sample in a furnace operating at 1300 °C or higher. A suitable elemental analyzer is the LECO CHNS-932 analyzer with a VTF-900 high température pyrolysis furnace.

Generally, oxidation of a material occurs in an oxidizing environment. For example, the oxidation can bc effected or aided by pyrolysis in an oxidizing environment, such as in air or argon enriched in air. To aid in the oxidation, various chemîcal agents, such as oxidants, acids or bases can be added to the material prior to or during oxidation. For example, a peroxide (e.g., benzoyl peroxide) can be added prior to oxidation.

Some oxidative methods of reducing recalcitrance in a biomass feedstock employ Fcnton-type chemistry. Such methods are disclosed, for example, in U.S. Serial No. 12/639,289, the complète disclosure of which is incorporated herein by référence.

Exemplary oxidants include peroxides, such as hydrogen peroxide and benzoyl peroxide, persulfates, such as ammonium persulfate, activated forms of oxygen, such as ozone, permanganates, such as potassium permanganate, perchlorates, such as sodium perchlorate, and hypochlorites, such as sodium hypochlorite (household bleach).

In some situations, pH is maintained at or below about 5.5 during contact, such as between 1 and 5, between 2 and 5, between 2.5 and 5 or between about 3 and 5. Oxidation conditions can also include a contact period of between 2 and 12 hours, e.g., between 4 and 10 hours or between 5 and 8 hours. In some instances, température is maintained at or below 300 °C, e.g., at or below 250,200, 150,100 or 50 °C. In some instances, the température romains substantially ambient, e.g., at or about 20-25 °C.

In some embodiments, the one or more oxidants are applied as a gas, such as by generating ozone in-situ by irradiating the material through air with a bcam of particles, such as électrons.

In some embodiments, the mixture further includes one or more hydroquinones, such as 2,5-dimethoxyhydroquinone (DMHQ) and/or one or more benzoquinones, such as 2,5-dimethoxy-l,4-benzoquinone (DMBQ), which can aid in électron transfer réactions.

In some embodiments, the one or more oxidants are electrochemically-generated in-situ. For example, hydrogen peroxide and/or ozone can be electro-chemically produced within a contact or réaction vcssel.

Other Processes To Solubilize, Red u ce Recalcitrance Or To Functionalîze

Any of the processes of this paragraph can be used alone without any of the processes described herein, or in combination with any of the processes described herein (in any order): steam explosion, chemical treatment (e.g., acid treatment (including concentrated and dilute acid treatment with minerai acids, such as sulfuric acid, hydrochloric acid and organic acids, such as trifluoroacetic acid) and/or base treatment (e.g., treatment with lime or sodium hydroxide)), UV treatment, screw extrusion treatment (see, e.g., U.S. Serial No. 13/099,151, solvent treatment (e.g., treatment with ionic liquids) and freeze milling (see, e.g., U.S. Serial No. 12/502,629 now U.S. Patent No. 7,900,857).

PRODUCTION OF FUELS. ACIDS, ESTERS AND/OR OTHER PRODUCTS

AND USES

A typical feedstock obtained at least in part from plants contains cellulose, hemiccllulose, and lignin plus lesser amounts of proteins, cxtractables and minerais. After onc or more of the processing steps discusscd above hâve been performed on the feedstock, the complex carbohydrates containcd in the cellulose and hemicellulose fractions can in some cases bc proccsscd into fcrmcntablc sugars, optionally, along with acid or enzymatic hydrolysis. The sugars liberated can be converted into a variety of products, such as alcohols or organic acids. The product obtained dépends upon the microorganism utilized and the conditions under which the bioproccssing occurs. In other embodiments, the treated feedstock can be subjected to thermochemical conversion, 30 or other processing.

»

Examples of methods of further processing the treated feedstock are discussed in the following sections.

Saccharification

In order to convert the treated feedstock to a form that can be readily fermented, in some implémentations the cellulose in the feedstock is first hydrolyzcd to low molecular weight carbohydrates, such as sugars, by a saccharifying agent, e.g., an enzyme, a process referred to as saccharification. In some implémentations, the saccharifying agent comprises an acid, e.g., a minerai acid. When an acid is used, coproducts may be generated that are toxic to microorganisms, in which case the process can further include removing such co-products. Removal may be performed using an activated carbon, e.g., activated charcoal, or other suitable techniques.

The treated feedstock can be hydrolyzcd using an enzyme, e.g., by combining the material and the enzyme in a solvent, e.g., in an aqueous solution.

Enzymes and biomass-destroying organisme that break down biomass, such as the cellulose and/or the lignin portions of the feedstock, contain or manufacture various ccllulolytic enzymes (ccllulascs), lîgninases or various small molécule biomassdestroying métabolites. These enzymes may be a complex of enzymes that act synergistically to dégrade crystalline cellulose or the lignin portions of biomass. Examples of cellulolytic enzymes include: endoglucanases, ccllobiohydrolases, and cellobiases (β-glucosidases). A cellulosic substratc is initially hydrolyzed by endoglucanases at random locations producing oligomeric intermediates. These intermediates are then substrates for cxo-splitting glucanases such as ccllobiohydrolase to produce cellobiose from the ends of the cellulose polymer. Cellobiose is a water-soluble l ,4-linked dimer of glucose. Finally cellobiase cleaves cellobiose to yield glucose.

Fermentation

Microorganisms can produce a number of useful intermediates and products by fermenting a low molecular weight sugar produced by saccharifying the treated feedstock. For example, fermentation or other bioprocesses can produce alcohols, organic acids, hydrocarbons, hydrogen, proteins or mixtures of any of these materials.

«

Yeast and Zymomonas bacteria, for example, can be used for fermentation or conversion. Other microorganisms are discussed in the Materials section, below. The optimum pH for fermentations is about pH 4 to 7. The optimum pH for yeast is from about pH 4 to 5, while the optimum pH for Zymomonas is from about pH 5 to 6. Typical fermentation times are about 24 to 168 (e.g., 24-96 hrs) hours with températures in the range of 20 °C to 40 °C (e.g., 26 °C to 40 °C), however thcrmophilic microorganisms prefer higher températures.

In some embodiments e.g., when anaérobie organisme arc used, at least a portion of the fermentation is conductcd in the absence of oxygen e.g., under a blanket of an inert gas such as N₂, Ar, He, CO₂ or mixtures thereof. Additionally, the mixture may have a constant purge of an inert gas flowing through the tank during part of or ail of the fermentation. In some cases, anaérobie condition can be achieved or maintained by carbon dioxide production during the fermentation and no additional inert gas is needed.

In some embodiments, ail or a portion of the fermentation process can be interrupted before the low molecular weight sugar is completely converted to a product (e.g. éthanol). The intermediate fermentation products include high concentrations of sugar and carbohydrates. The sugars and carbohydrates can be isolated as discussed below. These intermediate fermentation products can bc used in préparation of food for human or animal consumption. Additionally or aiternatively, the intermediate fermentation products can be ground to a fine particle size in a stainless-steel laboratory mill to producc a flour-like substance.

The fermentations include the methods and products that are disclosed in U.S. Provisional Application Serial No. 61/579,559, filed Dcccmber, 2011 and U.S. Provisional Application Serial No. 61/579,576, filed Dccember, 2011 incorporated herein by reference.

Mobile fermentons can be utilized, as described in U.S. Provisional Patent Application Serial No. 60/832,735, now Publishcd International Application No. WO 2008/011598. Similarly, the saccharification equipment can bc mobile. Further, saccharification and/or fermentation may bc performed in part or entirely during transit.

Fuel Cells

Where the methods described herein produce a sugar solution or suspension, this solution or suspension can subsequently be used in a fuel cell. For example, fuel cells utilizing sugars derived from cellulosic or lignocellulosic materiais are disclosed in U.S. Provisional Application Serial No. 61/579,568, filed December 22,2011, the complété disclosure of which is incorporated herein by reference.

Thermochemical Conversion

Thcrmochemical conversion can be performed on the treated feedstock to produce one or more desired intermediates and/or products. A thermochemical conversion o process includes changing molecular structures of carbon-containing material at elevated températures. Spécifie examples include gasificatîon, pyrolysis, reformation, partial oxidation and mixtures of these (in any order).

Gasification couverts carbon-containing materiais into a synthesis gas (syngas), which can include methanol, carbon monoxide, carbon dioxide and hydrogen. Many 15 microorganisms, such as acetogcns or homoacctogens are capable of utilizing a syngas from the thermochemical conversion of biomass, to produce a product that includes an alcohol, a carboxylic acid, a sait of a carboxylic acid, a carboxylic acid ester or a mixture of any of these, Gasification of biomass (e.g., cellulosic or lignocellulosic materiais), can be accomplishcd by a variety of techniques. For example, gasification can be accomplished utilizing staged steam reformation with a fluidized-bed reactor in which the carbonaceous material is first pyrolyzed in the absence of oxygen and then the pyrolysis vapors are reformed to synthesis gas with steam providing added hydrogen and oxygen. In such a technique, process heat cornes from burning char. Another technique utilizes a screw auger reactor in which moisture and oxygen are introduced at the pyrolysis stage and the process heat is generated from burning some of the gas produced in the latter stage. Another technique utilizes entrained flow reformation in which both cxtcmal steam and air are introduced in a single-stage gasification reactor. In partial oxidation gasification, pure oxygen is utilized with no steam,

POST-PROCESSING

Distillation

After fermentation, the resulting fluids can be distilled using, for example, a “beer column” to separate éthanol and other alcohols from the majority of water and residual solids. The vapor exiting the beer column can be, e.g., 35% by weight éthanol and can be fed to a rectification column. A mixture of nearly azcotropic (92.5%) cthanol and water from the rectification column can bc purified to pure (99.5%) cthanol using vapor-phasc molecular sieves. The beer column bottoms can bc sent to the first effect of a thrce-effect evaporator. The rectification column reflux condenser can provide heat for this first effect. After the first effect, solids can be separated using a centrifuge and dried in a rotary dryer. A portion (25%) of the centrifuge effluent can be recycled to fermentation and the rest sent to the second and third evaporator effects. Most of the evaporator condensate can be retumed to the process as fairly clcan condensate with a small portion split off to waste water treatment to prevent build-up of low-boiling compounds.

Other Possible Processine of Sugars

Proccssing during or after saccharification can include isolation and/or concentration of sugars by chromatography e.g., simulated moving bed chromatography, précipitation, centrifugation, crystal lization, solvent évaporation and combinations thereof. In addition, or optionally, processing can include isomerization of one or more of the sugars in the sugar solution or suspension. Additionally, or optionally, the sugar solution or suspension can be chemically processed e.g., glucose and xylose can be hydrogenated to sorbitol and xylitol respectively. Hydrogénation can bc accomplished by use of a catalyst e.g., Pt/y-AhCh, Ru/C, Raney Nickel in combination with H₂ under high pressure e.g., 10 to 12000 psi.

Some possible proccssing steps arc disclosed in in U.S. Provisional Application Serial No. 61/579,552, filed December 22, 2011, and in U.S. Provisional Application Serial No. 61/579,576 , filed December 22, 2011, incorporated by référencé above.

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INTERMEDIATES AND PRODUCTS

Using, e.g., such primary processes and/or post-processing, the treated biomass can be converted to one or more products, such as energy, fuels, foods and materials. Spécifie examples of products include, but are not limited to, hydrogen, sugars (e.g., glucose, xylose, arabinose, mannose, galactose, fructose, disaccharides, oligosaccharides and polysaccharides), alcohols (e.g., monohydric alcohols or dihydric alcohols, such as cthanol, n-propanol, isobutanol, .vec-butanol, tert-butanol or n-butanol), hydrated or hydrous alcohols, e.g., containing greater than 10%, 20%, 30% or even greater than 40% water, sugars, biodiescl, organic acids (e.g., acetic acid and/or lactic acid), hydrocarbons, co-products (e.g., proteins, such as cellulolytic proteins (enzymes) or single cell proteins), and mixtures of any of these in any combination or relative concentration, and optionally in combination with any additives, e.g., fuel additives. Other examples include carboxylic acids, such as acetic acid or butyric acid, salts of a carboxylic acid, a mixture of carboxylic acids and salts of carboxylic acids and esters of carboxylic acids (e.g., methyl, ethyl and n-propyl esters), ketones, aldéhydes, alpha, beta unsaturated acids, such as acrylic acid, olefins, such as ethylene, and mixtures of any of these. Other alcohols and alcohol dérivatives include propanol, propylene glycol, 1,4-butanediol, 1,3propancdiol, sugar alcohols (e.g., crythritol, glycol, glyccrol, sorbitol thrcitol, arabitol, ribitol, mannitol, dulcitol, fucitol, iditol, isomalt, maltitol, lactitol, xylitol and other polyols), methyl or ethyl esters of any of these alcohols, Other products include methyl acrylate, methylmethacrylate, lactic acid, propionic acid, butyric acid, succinic acid, 3hydroxypropionic acid, a sait of any of the acids and a mixture of any of the acids and respective salts.

In some embodiments using, e.g., such primary processes and/or post-processing, the treated biomass can be converted to a platform chemical. For exemple, as stated above, the treated biomass can bc converted to butanols (e.g., isobutanol, sec-butanol, tcrt-butanol or n-butanoi) which arc important platform chcmicals. For exemple, déhydration of butanols can producc butcncs such as 1-butcnc, cw-2-butcnc, trans-2butene and isobutene, which are highly valuable starting materials for synthetic fuels, lubricants and other valuable chemicals. Specifically, 1-butene can be used in the créations of polymers, e.g., linear low density polyethylene, 2-butene isomers are » · · valuable starting materials for lubricants and agricultural chemicals, and Isobutene can be polymerized to butyl rubber, methyl tert-butyl ether and isooctane. In addition, synthetic petroleum kerosene can be synthesized by oligomérization of butenes. Other intermediates and products, including food and pharmaceutical products, for example edible materials selected from the group consisting of pharmaceuticals, nutriceuticals, proteins, fats, vitamine, oils, fiber, minerais, sugars, carbohydrates and alcohols, are described in U.S. Serial No. 12/417,900, the full disclosure of which is hcrcby incorporatcd by reference herein.

MATERIALS

Modifîed Plant Materials

The plant feedstock is obtained at least in part from one or more types of modifîed plants, as dîscussed herein. In some cases, the feedstock includes more than one type of plant, and/or more than one portion of the plant, e.g., the stalk, fruit, and cob of a com plant. The plant may be, for example, a com, soybean, beet, cotton, rapeseed, potato, rice, alfalfa, or sugarcanc plant. The plant may also be any of the many types of genetically modifîed plants that are grown. The feedstock may contain a mixture of different types of plants, different parts of a particular plant, and/or mixtures of plant materials with other materials e.g., biomass materials.

In some cases the entire plant can be used. For cxample, in cases where a crop is ruined by adverse growing conditions (e.g., drought, frost, flooding, pest infestation) the ruined crop can be useful in the methods and processcs described herein.

Other Feedstock Materials

In addition or as an alternative to the modifîed plant materials dîscussed above, the feedstock can include other materials e.g., bïomass materials, that may or may not be genetically modifîed. The biomass can bc, e.g., a cellulosic or lignocellulosic material. Such materials include paper and paper products (e.g., polycoated paper and Kraft paper), wood, wood-related materials, e.g., particle board, grasses, rice hulls, bagasse, jute, hemp, flax, bamboo, sisal, abaca, straw, switchgrass, alfalfa, hay, com cobs, corn stover,

coconut haïr; and materials high in α-cellulose content, e.g., cotton. Feedstocks can be obtained from virgin scrap textile materials, e.g., rcmnants, post consumer waste, e.g., rags. When paper products are used they can be virgin materials, e.g., scrap virgin materials, or they can be post-consumer waste. Aside from virgin raw materials, post5 consumer, industrial (e.g., offal), and processing waste (e.g., effluent from paper processing) can also be used as fiber sources. Biomass feedstocks can also be obtained or derived from human (e.g., sewage), animal or plant wastes. Additional cellulosic and lignocellulosic materials hâve been described in U.S. Patent Nos. 6,448,307; 6,258,876; 6,207,729; 5,973,035 and 5,952,105.

In some embodiments, the biomass material includes a carbohydrate that is or includes a material having one or more p-l,4-linkagcs and having a number average molecular weight between about 3,000 and 50,000. Such a carbohydrate is or includes cellulose (I), which is derived from (β-glucose 1) through condensation of β(1,4)~ glycosidic bonds. This linkage contrasts itself with that for a(l,4)-glycosîdic bonds présent in starch and other carbohydrates.

fl

I «

I

Starchy materials include starch itself, e.g., corn starch, wheat starch, potato starch or rice starch, a dérivative of starch, or a material that includes starch, such as an edible food product or a crop. For example, the starchy material can be arracacha, buckwheat, banana, barley, cassava, kudzu, oca, sago, sorghum, regular household potatocs, swcet potato, taro, yams, or one or more beans, such as favas, lentils or peas. Blends of any two or more starchy materials are also starchy materials.

In some instances the biomass is a microbial material. Microbial sources include, but arc not limited to, any naturally occurring or gcnctîcally modified microorganism or organism that contains or is capable of providing a source of carbohydrates (e.g., cellulose), for exemple, protists, e.g., animal protists (e.g., protozoa such as flagellâtes, amoeboids, ciliatcs, and sporozoa) and plant protists (e.g., algae such alveolates, chlorarachniophytes, cryptomonads, cuglenids, glaucophytes, haptophytes, red algae, stramenopiles, and viridaeplantae). Other examples include seaweed, plankton (e.g., macroplankton, mesoplankton, microplankton, nanoplankton, picoplankton, and femptoplankton), phytoplankton, bactcria (e.g., gram positive bacteria, gram négative bacteria, and extrcmophiles), yeast and/or mixtures of these. In some instances, microbial biomass can be obtained from natural sources, e.g., the océan, lakes, bodies of water, e.g., sait water or fresh water, or on land. Alternatively or in addition, microbial biomass can be obtained from culture Systems, e.g., large scale dry and wet culture Systems.

Saccharifving Agents

a

Suitable enzymes include cellobiases and cellulases capable of degrading biomass.

Suitable cellobiases include a cellobiase from Aspergillus niger sold under the tradename NOVOZYME 188™.

Cellulases are capable of degrading biomass, and may be of fungal or bacterial origin. Suitable enzymes include cellulases from the généra Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, Chrysosporium and Trichoderma, and include species of Humicola, Coprinus, Thielavia, Fusarium, Myceliophthora, Acremonium, Cephalosporium, Scytalidium, Pénicillium or Aspergillus (see, e.g., EP 458162), especially those produced by a strain selected from the species Humicola insolens (reclassified as Scytalidium thermophilum, see, e.g., U.S. Patent No. 4,435,307), Coprinus cinereus, Fusarium oxysporum, Myceliophthora thermophila, Meripilus giganteus, Thielavia terrestris, Acremonium sp., Acremonium persicinum, Acremonium acremonium, Acremonium brachypenium, Acremonium dichromosporum, Acremonium obclavatum, Acremonium pinkertoniae, Acremonium roseogriseum, Acremonium incoloratum, and Acremonium furatum', preferably from the species Humicola insolens DSM 1800, Fusarium oxysporum DSM 2672, Myceliophthora thermophila CBS 117.65, Cephalosporium sp. RYM-202, Acremonium sp. CBS 478.94, Acremonium sp. CBS 265.95, Acremonium persicinum CBS 169.65, Acremonium acremonium AHU 9519, Cephalosporium sp. CBS 535.71, Acremonium brachypenium CBS 866.73, Acremonium dichromosporum CBS 683.73, Acremonium obclavatum CBS 311.74, Acremonium pinkertoniae CBS 157.70, Acremonium roseogriseum CBS 134.56, Acremonium incoloratum CBS 146.62, and Acremonium furatum CBS 299.70H. Ccllulolytic enzymes may also be obtained from Chrysosporium, preferably a strain of Chrysosporium lucknowense. Additionally, Trichoderma (particularly Trichoderma viride, Trichoderma reesei, and Trichoderma koningii), alkalophilîc Bacillus (sce, for exampic, U.S. Patent No. 3,844,890 and EP 458162), and Streptomyces (scc, e.g., EP 458162) may bc used.

Enzyme complexes may bc utilized, such as those available from Gcncncorc under the tradename ACCELLERASE®, for example, Accellerase® 1500 enzyme complex. Accellerase 1500 enzyme complex contains multiple enzyme activities, mainly exoglucanase, endoglucanase (2200-2800 CMC U/g), hemi-cellulase, and beta-

glucosidasc (525-775 pNPG U/g), and has a pH of 4.6 to 5.0. The endoglucanase activity of the enzyme complex is exprcssed in carboxymethylcellulose activity units (CMC U), while the beta-glucosidase activity is reported in pNP-glucoside activity units (pNPG U).

In one embodiment, a blend of Accelierase® 1500 enzyme complex and NOVOZYME™ 5 188 cellobiase îs used.

Fermentation Agents

The microorganism(s) used in fermentation can bc naturel microorganisms and/or engineered microorganisms. For example, the microorganîsm can be a bacterium, e.g., a cellulolytic bacterium, a fungus, e.g., a yeast, a plant or a protist, e.g., an algae, a protozoa or a fungus-like protist, e.g., a slime mold. When the organisme are compatible, mixtures of organisms can be utilized.

Suitable fermenting microorganisms have the ability to convert carbohydrates, such as glucose, fructose, xylosc, arabînose, mannose, galactose, oligosaccharides or 15 polysaccharides into fermentation products. Fermenting microorganisms include strains of the genus Sacchromyces spp. e.g., Sacchromyces cerevisiae (baker’s yeast), Saccharomyces distaticus, Saccharomyces uvarunv, the genus Kluyveromyces, e.g., spccics Kluyveromyces marxianus, Kluyveromyces fragilis', the genus Candida, e.g., Candidapseudotropicalis, and Candida brassicae, Pichia stipitis (a relative of Candida shehatae, the genus Clavispora, e.g., species Clavispora lusitaniae and Clavispora opuntiae, the genus Pachysolen, e.g., species Pachysolen tannophilus, the genus Bretannomyces, e.g., species Bretannomyces ciausenii (Philippidis, G. P., 1996, Cellulose bioconversion technology, in Handbook on Biocthanol: Production and Utilization, Wyman, C.E., ed., Taylor & Francis, Washington, DC, 179-212). Other suitable microorganisms include, for example, Zymomonas mobilis, Clostridium thermocellum (Philippidis, 1996, supra), Clostridium saccharobutylacetonicum, Clostridium saccharobutylicum, Clostridium Puniceum, Clostridium beijernckii, Clostridium acetobutylicum, Moniliella pollinis, Yarrowia lipolytica, Aureobasidium sp., Trichosporonoides sp., Trigonopsis variabilis, Trichosporon sp., Moniliellaacetoabutans,

Typhula variabilis, Candida magnoliae, Ustilaginomycetes, Pseudozyma tsukubaensis, yeast species of généra Zygosaccharomyces, Debaryomyces, Hansenula and Pichia, and fiingi of the dematioid genus Torula.

Commercially available yeasts include, for example, Red Star®/Lesaffre Ethanol Red (available from Red Star/Lesaffre, USA), FALI® (available from Fleischmann’s 5 Yeast, a division of Bums Philip Food Inc., USA), SUPERSTART® (available from Alltech, now Lalemand), GERT STRAND® (available from Gert Strand AB, Sweden) and FERMOL® (available from DSM Specialtics).

OTHER EMBODIMENTS

A number of embodiments of the invention have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention.

For example, the process parameters of any of the processing steps discussed herein can be adjusted based on the lignin content of the feedstock, for example as 15 disclosed in U.S. Serial No. 12/704,519, the full disclosure of which is incorporated herein by reference.

The process may include any of the features described in U.S. Application Serial No. 13/276,192, the full disclosure of which is incorporated herein by reference, including treating a ccllulosic or lignoccllulosic material to altcr the structure of the material by irradiating the material with relativeiy low voltage, high power électron beam radiation, boiling or steeping the feedstock prior to saccharification, and irradiating a ccllulosic or lignocellulosic material with an électron beam at a dose rate of at least 0.5 Mrad/sec.

While it is possible to perform ail the processes described herein at one physicaï location, in some embodiments, the processes arc completed at multiple sites, and/or may be performed during transport.

Lignin lîbcratcd in any process described herein can bc capturcd and utilizcd. For cxample, the lignin can bc used as capturcd as a plastic, or it can be synthetically upgradcd to other plastics, ln some instances, it can bc utilizcd as an energy source, e.g., 30 bumed to provide heat. In some instances, it can also be converted to lignosulfonates, which can be utilized as bïnders, dispersants, emulsifiers or as séquestrants.

. ¹

Measurement of the lignin content of the starting feedstock can be used in process control in such lignin-capturing processes.

When used as a binder, the lignin or a lignosulfonate can, e.g., be utilized in coal briquettes, in ceramics, for binding carbon black, for binding fertilizers and herbicides, as a dust suppressant, in the making of plywood and particle board, for binding animal feeds, as a binder for fibergiass, as a binder in linoléum paste and as a soil stabilizer.

As a dispersant, the lignin or lignosulfonates can bc used, e.g., concrète mixes, clay and ceramics, dycs and pigments, Icathcr tanning and in gypsum board.

As an emulsifier, the lignin or lignosulfonates can be used, e.g., in asphalt, pigments and dyes, pesticides and wax émulsions.

As a séquestrant, the lignin or lignosulfonates can be used, e.g., in micro-nutrient Systems, cleaning compounds and water treatment Systems, e.g., for boiler and cooling Systems.

As a heating source, lignin generally has a higher energy content than holocelhilose (cellulose and hemicellulose) since it contains more carbon than homocellulosc. For example, dry lignin can hâve an energy content of between about 11,000 and 12,500 BTU per pound, compared to 7,000 an 8,000 BTU per pound of holoccllulosc. As such, lignin can bc dcnsificd and convcrtcd into briquettes and pellets for buming. For example, the lignin can be converted into pellets by any method described herein. For a slower buming pellet or briquette, the lignin can be crosslinked, such as applying a radiation dose of between about 0.5 Mrad and 5 Mrad. Crosslinking can make a slower buming form factor, The form factor, such as a pellet or briquette, can be converted to a “synthetic coal” or charcoal by pyrolyzing in the absence of air, e.g., at between 400 and 950 °C. Prior to pyrolyzing, it can be désirable to crosslînk the lignin to maintain structural integrity.

Accordingly, other embodiments are within the scope of the foliowing daims.

EXAMPLES OF GENETICALLY MODIFIED PLANTS

The foliowing US Patents and US Patent applications disclose, by example, genetically modified material (e.g., plants, parts of plants) for the proccsscs described herein or together with any materials described herein.

7566817	7763783	7714209	7659459	7615694	7534943
7652202	7763782	7714208	7659458	7615693	7531724
7569747	7763780	7709712	7659457	7615692	7528305
7405344	7759563	7709711	7659456	7612268	7528304
7683237	7759562	7709710	7659455	7612267	7525029
7615621	7759561	7709709	7655849	7612266	7525027
7816591	7759560	7709708	7655847	7612260	7525026
7816590	7759559	7705221	7655846	7608765	7521614
7816589	7750215	7705220	7655845	7608763	7521613
7816587	7745707	7705216	7655844	7608762	7521612
7807904	7741547	7700859	7655841	7605316	7521609
7807903	7741546	7700858	7642433	7605315	7521607
7807902	7737348	7700857	7642432	7605314	7518044
7807901	7737347	7692077	7642431	7605313	7518043
7807900	7737346	7682076	7642430	7605312	7518042
7807899	7737345	7687689	7642429	7605311	7518041
7807898	7737344	7683243	7642428	7605309	7514612
7807897	7737343	7683242	7638694	7601900	7514611
7807896	7732685	7683241	7638693	7601899	7514610
7807895	7732684	7683239	7638692	7598441	7514609
7807894	7728208	7678978	7638691	7598440	7511204
7807893	7723589	7678975	7638690	7595440	7511203
7807892	7723588	7678974	7638689	7595439	7511202
7807891	7723587	7678973	7632995	7595438	7511201
7807890	7723588	7678972	7632994	7595437	7511200
7807889	7723585	7678971	7632990	7592527	7507880
7807880	7718870	7678970	7629519	7592526	7507879
7807887	7718869	7678969	7629518	7592525	7504569
7804011	7718868	7678968	7629517	7592521	7504567
7804010	7718867	7678967	7629516	7592520	7504566
7804009	7718866	7678966	7629515	7582434	7501565
7804008	7718865	7674961	7626101	7576265	7501564
7804007	7718864	7671256	7626100	7566822	7498494
7804006	7718863	7667113	7626099	7563966	7495157
7804005	7718862	7667112	7626098	7563965	7495156
7804004	7718861	7667111	7622680	7560625	7495155
7804003	7718860	7667110	7622659	7557279	7488874
7804002	7718859	7667109	7619153	7650655	7488873
7804001	7714216	7663037	7619152	7547827	7488872
7804000	7714215	7663036	7619151	7547826	7485783
7803999	7714214	7663035	7619150	7547824	7479589
7786359	7714213	7663034	7619147	7544868	7479586
7781651	7714212	7659462	7615697	7544867	7479585
7781650	7714211	7659461	7615696	7541527	7476785
7772469	7714210	7659460	7615695	7541523	7476784

7476703	7301070	7262348	7179972	7737332	7544863
7473Θ30	7301009	7259303	7179969	7732879	7544862
7473Θ29	7301860	7256335	7176365	7728206	7534939
7473826	7375266	7256334	7173172	7718854	7531718
7470639	7375265	7256333	7169983	7714202	7528246
7470036	7371946	7250564	7166778	7714201	7521594
7460477	7371947	7247777	7166777	7709706	7511130
7462765	7371946	7241944	7166776	7709705	7465849
7462764	7371945	7235728	7164068	7388135	7423203
7462763	7368643	7235725	7164065	7388134	7417177
7459613	7360640	7235724	7161070	7381861	7417176
7459612	7365252	7235722	7161069	7253345	7408096
7459611	7365251	7235720	7157630	7250583	7405343
7456345	7365250	7232945	7157626	7247774	7385107
7456344	7365249	7232944	7157625	7223907	7365241
7456343	7361020	7232943	7157624	7189514	7335812
7456342	7361819	7230173	7157281	RE39.580	7329799
7453031	7361810	7227062	7154031	7816581	7304206
7453030	7361817	7227061	7151208	7812219	7294711
7449622	7361015	7223908	7148410	7807874	7288408
7449621	7361814	7220900	6906250	7807873	7268276
7449620	7350427	7217074	6864409	7807812	7262339
7449619	7355107	7217873	6855877	7007811	7250501
7439424	7351890	7217872	6825400	7803928	7244877
7432427	7351Θ8Θ	7217871	6114610	7799970	7230165
7432426	7342156	7211717	6103959	7790953	7227056
7429696	7342155	7211716	6103958	7706353	7217867
7423207	7342152	7208663	6084161	7788350	7217865
7423206	7339101	7208662	6054640	7750207	7205457
7423204	7339100	7208661	7112725	7745694	7195917
7417103	7339099	7208660	7825304	7728190	7186893
74171Θ2	7339098	7205466	7825303	7714189	7157619
7414101	7335827	7205465	7825302	7705201	7151204
7408099	7335826	7205464	7025301	7700838	7148398
7399915	7335022	7199291	7025300	7692067	7141722
7399912	7329803	7199290	7025300	7874952	7138278
7399911	7321088	7193146	7820888	7674894	7122719
7394003	7321087	7193143	7820887	7662940	7112717
7390946	7321086	7109906	7803997	7655838	7078592
7390945	7321085	7109904	7799972	7635764	7067722
7388140	7321084	7189903	7750213	7625738	7064249
7388139	7319182	7186906	7750212	7615680	7022897
7385122	7317155	7186904	7745704	7805244	6943281
7305121	7297848	7186903	7741543	7601890	8916970
7305120	7294772	7186901	7737335	7595382	6841717
7381874	7208704	7186899	7737334	7589188	6822142
7301873	7268279	7183471	7737333	7553952	8803501

6620988	7728196	7626089	7557277	7491870	7301867
6538179	7723583	7626088	7557276	7488069	7381866
6538178	7714198	7626087	7557275	7488868	7378570
6501009	7709703	7626086	7557274	7488867	7370577
6476295	7705211	7622646	7557273	7485701	7375262
6448476	7705208	7622645	7557272	7485780	7371938
6448473	7705207	7622644	7557271	7482516	7368637
6264949	7700849	7622643	7557270	7479583	7368635
6281016	7700847	7622642	7554016	7479582	7350420
6177615	7700846	7619143	7554015	7468474	7355103
6175061	7700844	7619142	7554014	7459609	7355102
6156573	7700843	7619141	7554013	7453029	7351886
6107549	7692070	7619140	7550653	7453028	7351885
6023013	7687686	7619139	7531722	7446244	7345228
5463175	7667685	7615688	7531721	7442864	7345227
7531725	7687684	7615687	7531720	7442863	7345226
7468476	7678965	7612259	7531719	7442862	7345225
7253346	7678964	7608761	7528306	7442860	7345224
7214863	7678963	7605306	7528301	7439422	7342151
7186900	7676962	7598434	7525020	7423200	7342150
7166780	7659454	7595435	7525025	7423199	7332656
7166779	7659453	7592517	7525019	7414177	7332655
7157628	7659452	7592516	7525018	7414176	7329001
7157627	7655839	7592514	7525017	7408097	7326832
7563949	7652199	7592513	7521608	7405349	7321082
7807684	7652198	7592512	7521605	7405348	7321079
7799973	7652197	7592511	7518036	7399909	7314983
7790964	7649129	7562810	7514607	7399907	7314982
7786357	7649128	7579525	7514606	7396983	7314981
7781649	7649127	7579524	7514605	7394000	7314980
7777104	7649126	7579523	7514604	7390942	7312382
7777103	7642413	7572960	7514603	7390941	7312380
7767887	7642412	7572958	7514602	7390940	7309818
7759556	7642411	7572957	7507878	7390939	7307201
7759553	7642410	7572956	7507877	739093Θ	7304218
7759551	7642409	7569752	7504565	7388132	7304214
7732677	7642408	7569751	7504564	7388131	7304213
7732676	7642407	7569750	7501563	7388130	7301076
7732675	7632987	7566821	7501562	7385117	7297843
7732674	7632985	7566820	7501560	7385116	7294770
7732673	7629510	7563955	7490491	7385115	7294768
7732672	7629509	7563954	7498490	7385113	7294765
7728204	7629508	7563953	7498489	7385112	7294764
7728203	7629507	7560619	7498486	7385111	7294763
7728202	7629506	7560618	7498485	7385110	7291771
7728201	7626091	7560617	7498484	7385109	7291769
7728199	7628090	7580616	7491871	7385108	7285704

7279621	7132591	7045687	6900373	6080916	5902923
7278648	7129399	7045886	6900372	6063990	5898100
7271324	7126046	7041881	6894207	6063989	5880346
7265277	7119260	7041880	6888049	6051761	5880345
7265276	7119259	7041879	6888048	6043414	5880344
7265275	7119258	7038114	6884927	6040499	5872304
7265274	7115801	7034210	6884926	6037529	5872303
7265273	7109399	7034209	6884925	6034303	5866774
7265272	7105728	7030301	6884924	6034302	5866773
7265271	7105727	7030300	6884923	6034301	5866772
7265270	7105726	7019199	6881879	6034300	5866771
7259299	7102062	7012174	6875908	6034299	5859352
7259298	7102081	7005583	6870079	6031159	5723745
7256330	7098385	7002062	6861579	6020542	7268274
7247772	7098384	7002058	6858784	6020541	7402731
7247771	7091403	6982367	6858783	6018108	6865556
7244881	7087815	6982366	6852912	6018107	5424412
7241941	7084328	6979761	6849786	6005171	5463175
7241939	7084326	6979760	6849785	6005170	5484956
7235718	7084325	6972355	6846973	6002073	5554798
7217870	7084324	6972354	6835873	5998709	559387
7217869	7081572	6972353	6828409	5998708	5641876
7217868	7078600	6969787	6815589	5998707	5659122
7196253	7078598	6967263	68155ΘΘ	5998706	571084
7196252	7078597	6960707	6815587	5998705	5728925
7196251	7078595	6958436	6815586	5998704	5750871
7193140	7074989	6953876	6809237	5998703	5804425
7193139	7074988	6951973	6781040	5990391	5859347
7193137	7071390	6936754	6653534	5986179	6020190
7189900	7071389	6936753	6198027	5986178	6025545
7189898	7071388	6933423	6177618	5981851	6040497
7183467	7071387	6924418	6169227	5981850	6051753
7183465	7067723	6919498	6137034	5981849	6180774
7183464	7064253	6914174	6133510	5981848	6218188
7183463	7060878	6914173	6124527	5981845	6340593
7183462	7060877	6914172	6121518	5977449	6489542
7183461	7057096	6911585	6121517	5977448	6501009
7176359	7057095	6911581	6121516	5977447	6548291
7176358	7057094	6911580	6121515	5977444	6573240
7176357	7057093	6911579	6114604	5973235	6645497
7173168	7057092	6911578	6103957	5969218	6660911
7169976	7053280	6906248	6100454	5969217	6737273
7169975	7053279	6906247	6096949	5945588	6753463
7169974	7053272	6906246	6091005	5942666	6825400
7166774	7049494	6903253	6087562	5932786	6893872
7148408	7049493	6903251	6084159	5929310	6900371
7138570	7045691	6900376	6080918	5907088	6943282

6949696	7462510	7834247	7772465	7663031	7601894
6962705	7473819	7834246	7772370	7663029	7598443
7064249	7465850	7834245	7767889	7655848	7598442
7112665	7456337	7834240	7767888	7655843	7598439
7112725	7456335	7829764	7763778	7655842	7598438
7141722	7442853	7829760	7763465	7652201	7598437
7157281	7439417	7825310	7759564	7652200	7598435
7223907	7435875	7825309	7759555	7652195	7595436
7227056	7427698	7825308	7759554	7645923	7592524
7250501	7427696	7825307	7759544	7645922	7592523
7288643	7425666	7825299	7759543	7645921	7592522
7381861	7425665	7825294	7754949	7642421	7592519
7435807	7423196	7825234	7754948	7642420	7592505
7449564	7399904	7820895	7750216	7642419	7589264
7514544	7399903	7820894	7745706	7642418	7589263
RE38825	7375209	7820893	7745705	7642417	7589261
RE39247	7317140	7820892	7745702	7638695	7589260
7829761	7303919	7820891	7745701	7638688	7589259
7807882	7271316	7820886	7741545	7632993	7589258
7803987	7259294	7820885	7737342	7832992	7589257
7799971	7238856	7816586	7737341	7632989	7589178
7795500	7235713	7816585	7737340	7632988	7586028
7795414	7220585	7812231	7737336	7629514	7586027
7790873	7189893	7812230	7737330	7629513	7586026
7763777	7186561	7812226	7736897	7629512	7586025
7763776	7179962	7812225	7732683	7629511	7582816
7718858	7176026	7812223	7732668	7629505	7582815
7718857	7166767	7812216	7728207	7629504	7582614
7714190	7164057	7807883	7718856	7624533	7582813
7709698	7161063	7807876	7714205	7622647	7582812
7652203	7135618	7803998	7714187	7622637	7582811
7622570	7125719	7803996	7714184	7619149	7582808
7619137	7105723	7803993	7709702	7619148	7579530
7608750	7067261	7603990	7705219	7615690	7579520
7608757	7034208	7803989	7705218	7612265	7579522
7598431	6867351	7799977	7700856	7612264	7576271
7579517	6825399	7799975	7700855	7612263	7576270
7563948	6818805	7799974	7700854	7612262	7576269
7521598	6784338	7799566	7700836	7612256	7576268
7521597	6774288	7795508	7700832	7612254	7576267
7514599	6720477	7795506	7692061	7612251	7576266
7504559	6710229	7790969	7687687	7608764	7572963
7498482	6669939	7790874	7683240	7608755	7572982
7498429	6677504	7777107	7667115	7608752	7572961
7495151	6320518	7777106	7667107	7605307	7572955
7485775	6225526	7772468	7663033	7601898	7569757
7402511	7834257	7772467	7863032	7601897	7569756

7569755	7525023	7456339	7371936	7317149	7276650
7569754	7525022	7442881	7365253	7317148	7276849
7569753	7525021	7439425	7361812	7317147	7276647
7569749	7525020	7439421	7361807	7317146	7276596
7566619	7521611	7439348	7358425	7317145	7273975
7563964	7521610	7435885	7358424	7317143	7273973
7563963	7521604	7435883	7358423	7317137	7273972
7563962	7521603	7435881	7355108	7314990	7273971
7563961	7521602	7435880	7355106	7314989	7273965
7563960	7521601	7435879	7355105	7314988	7271327
7563959	7518037	7432424	7355104	7314987	7271326
756395Θ	7514601	7432423	7351882	7312385	7271323
7563957	7511205	7432422	7351878	7312384	7271319
7560624	7511196	7432421	7348469	7312377	7270380
7560623	7511195	7432418	7348468	7312375	7268278
7560612	7511194	7429695	7345230	7309816	7268277
7557266	7511193	7427702	7342157	7306948	7268270
7557263	7511192	7427701	7342154	7304222	7268226
7554020	7511188	7427700	7339097	7304221	7265279
7553951	7504568	7423202	7339096	7304212	7265265
7550657	7504558	7423197	7339092	7304211	7262350
7550656	7501561	7420103	7335828	7301082	7262349
7550575	7498488	7414180	7335825	7301080	7262347
7547832	7498487	7414179	7335824	7301079	7262346
7547831	7498413	7414174	7335823	7301075	7262345
7547830	7495154	7411118	7335817	7301069	7262342
7547829	7495150	7411113	7332660	7297850	7259305
7547825	7491869	7411112	7332659	7297849	7259304
7547822	7485779	7399914	7332658	7297841	7259302
7544869	7485778	7399910	7332650	7294774	7259301
7544866	7482515	7399908	7329806	7294769	7256332
7544865	7482513	7399908	7329805	7294767	7256331
7544864	7479588	7396980	7329804	7294766	7256322
7544857	7479581	7393999	7326836	7291774	7256280
7541526	7476781	7388141	7326835	7291773	7253000
7541525	7473828	7388137	7326833	7288703	7250552
7541524	7473827	7388133	7326830	7288701	7241943
7541521	7473821	7388128	7323623	7288700	7241942
7541520	7470838	7388125	7321089	7288699	7241940
7541517	7470834	7381872	7321083	7285707	7241934
7538261	7470833	7381871	7321031	7285706	7238859
7528308	7468278	7381865	7319183	7285702	7235723
7528307	7465858	7381863	7317154	7282629	7232946
7528300	7465852	7378574	7317153	7282627	7230172
7528299	7462766	7375264	7317152	7282626	7230171
7528293	7462760	7375263	7317151	7279615	7230169
7525024	7459610	7371944	7317150	7276652	7230158

V

7227065	7179971	7151207	7064255	7005565	6951974
7227064	7179970	7151205	7064252	7002061	8949699
7227063	7179968	7148406	7064251	7002056	6946589
7227060	7179967	7148401	7064250	6998518	6943279
7227059	7179963	7141721	7064247	6995305	6936756
7227056	7179955	7129402	7060879	6995304	6936755
7220902	7179599	7129401	7060813	6995303	6936752
7220901	7176364	7129395	7053286	6992240	6936751
7214Θ65	7176363	7122725	7053285	6992239	6933427
7214864	7176362	7115802	7053284	6992238	6933425
7214860	7176360	7112731	7053283	6992237	6930230
7214857	7176356	7112729	7053282	69Θ94Θ1	6930229
7214855	7176349	7112728	7053275	6989480	6930225
7214054	7176027	7109403	7049499	6989479	6927327
7214852	7173174	7109391	7049495	6989478	6927326
7211716	7173173	7109390	7045692	6989475	8924421
7211714	7169908	71020Θ4	7045682	8989474	6921852
7211712	7169987	7102063	7041887	6987217	6921850
7205455	7169906	7098390	7041886	6987212	6921847
7205453	7169905	7098301	7041874	6984778	6919500
RE39.562	7169984	7094957	7038109	6982371	6919499
7202403	7169980	7094956	7038108	6979764	6916975
7202402	7169979	7091407	7034214	6979763	6914178
7199294	7169978	7091406	7034213	6979759	6914177
7199293	7169977	7091398	7034211	6977327	6914171
7199292	7169973	7087823	7030303	6974900	6914170
7199289	7166764	7087822	7030302	6974899	6911587
7196256	7166782	7087821	7030290	6972357	6911577
7196255	7166781	7087820	7026533	6972356	6909039
7196254	7166769	7084335	7022904	6972352	6909038
7193145	7166765	7004332	7022902	6969790	6906251
7193144	7164070	7084327	7022099	6969789	6906243
7193141	7164069	7001566	7019200	6969780	6905857
7193136	7164067	7078603	7019198	6969706	6903254
7193135	7164066	7078602	7015386	6967269	6903205
7193130	7164063	7078601	7015385	6987268	6900370
7189905	7164062	7078596	7015381	6967267	6900377
7189Θ02	7164061	7078589	7015380	6967264	6097365
7189901	7164056	7074991	7015379	6965063	6897364
7189899	7161074	7071397	7015376	6960708	6097363
7169089	7161073	7071396	7015375	6958438	6897362
7106905	7161072	7071395	7012177	6958437	6897361
7186902	7161071	7071394	7012176	6956153	6897360
7186096	7161068	7071393	7009094	6956150	6891090
7183472	7161065	7067727	7009093	6953878	6891085
7183469	7157632	7067720	7009087	6953877	6888051
7103460	7154030	7064258	7005566	6951975	6887708

6681881	6815585	6759578	6706949	6555732	6333452
8881880	6815584	8759577	6700041	6555673	6333451
687ΘΘ65	6815583	6756530	6693231	6541684	6331661
6878864	6815578	6756529	6677503	6538177	6329579
6878863	6812304	6756528	6677502	6538176	6326530
6875907	6812383	8753464	6667427	6528704	6328529
6872874 '	6812380	6750384	6660907	6518487	6323402
6872873	6009242	6750380	6657107	6510483	6323401
6864411	6Θ09241	0747196	6646182	6515202	6323400
6864408	6009236	6747193	6639131	6504084	6323399
6864407	6006408	6743970	6639126	6504083	6323398
6861577	6806407	6740798	6635807	6504082	6323015
6858785	6806406	6740798	6630615	6479730	6320106
6858782	6806405	6740795	6630614	6476292	6320105
6858781	6806404	6737566	6627797	6472105	6316704
6858778	6806401	6737565	6617499	6444074	6316703
6855878	6803508	6737562	6617498	6441151	6316702
6855876	6803498	6737560	6613967	6433259	6316700
6855875	6800796	6734348	6613966	6429362	6313384
6855874	6800795	6734347	6613965	6426452	6313383
6855871	6797868	6734345	6613964	6423008	6313382
6852913	6797867	6734341	6613963	6423806	6313381
6849791	6797866	6730837	6610911	6410829	6313376
6849789	6797865	6730836	6610910	6407315	0313375
6849708	6797864	6730035	6608243	6403862	6310274
6849767	6797863	6730834	6608240	6403060	6307132
6846976	6797859	6730829	6605762	6399056	6307131
6846975	6794563	6727413	6605761	6392127	6303051
6846974	6791016	6727412	6605760	6392126	6297433
6844480	6784350	6727410	6605759	6388179	6297432
6838593	6784349	6723903	6605758	63Θ8171	6297426
6835877	6784347	6723902	6605757	6380169	6291745
6835875	6784341	6720487	6605756	6384302	6288310
6833498	6781043	6720486	6605755	6372961	6287843
6831215	6781042	6720481	6600095	6369301	6284953
6828493	6781041	6720478	6586659	6369300	6284950
6828490	6777599	6720475	6586657	6362400	6284948
6825405	6777598	6717040	6583343	6359201	6271439
6825404	6777597	6717039	6583342	6346657	6271437
6825397	6777596	6717038	6583341	6344603	6268553
6822144	6777590	6717037	6580018	6342659	6265646
6822140	6774290	6717036	6576819	6339186	6265636
6818813	6774209	6717033	6576814	6339144	6259005
6810811	6774202	6713666	6573433	6337100	6259004
6818809	6770802	6713665	6566589	6335476	6255090
681Θ80Θ	6765132	6706954	6566584	6335197	6248935
6815592	6759580	6706951	6563020	6333453	6242673

6242672	6156958	6111167	5990392	5902921	5792909
6235976	6153817	6107551	5990389	5900526	5792908
6235972	6153816	6107550	5986185	5900524	5792907
6232529	6147285	6107545	5986184	5895835	5783100
6232527	6147284	6100030	5986183	5889188	5773697
6229079	6147283	6096953	5981854	5866768	5773684
6229078	6143962	6096951	5977457	5866767	5773682
6229077	6143956	6096947	5977456	5866766	5770790
6229074	6143955	6093875	5977451	5866765	5767347
6229073	6143954	6091007	5977445	5861541	5767344
6225537	6140562	6091006	5973237	5859354	5767343
6225529	6140557	6087567	5973234	5859341	5767340
6222103	6137037	6087566	5982772	5859320	5763757
6222102	6137036	6087565	5962771	5859319	5763747
6222101	6133514	6087564	5959185	5859318	5763746
6215049	6133513	6087559	5955361	5859317	5763744
6211445	6133508	6084164	5952550	5859316	5763743
6211440	6130370	6084160	5952549	5859313	5763243
6211437	6127610	6080919	5948957	5852226	5750868
6211435	6127609	6080913	5945587	5852225	5750849
6211434	6127603	6077998	5945586	5850024	5750847
6211433	6127602	6077997	5942671	5850016	5750843
6198026	6127600	6077993	5942670	5850013	5750842
6197561	6124535	6075186	5942669	5850012	5750841
6194638	6124534	6075182	5942668	5850011	5750839
6194637	6124533	6072104	5942667	5850010	5750838
6191343	6124532	6069304	5939608	5850009	5750835
6166001	6124531	6057491	5939607	5850007	5750834
6188000	6124530	6054639	5936148	5844118	5750832
6184448	6124529	6040505	5936147	5844117	5750831
6184445	6124526	6037530	5936142	5844116	5750829
6184439	6121524	6037523	5936141	5841015	5741684
6180857	6121523	6028254	5936140	5827940	5736627
6180856	6121522	6028252	5929313	5824844	5731499
6180850	6121520	6025547	5929311	5824524	5731497
6177613	6121514	6020543	5929301	5817918	5731496
6177611	6118056	6018113	5920002	5811651	5731494
6175065	6118055	6018112	5917134	5811650	5731493
6175058	6118054	6018111	5917130	5811639	5731492
6169234	6118053	6018110	5917129	5811638	5731491
6166305	6118052	6018109	5917125	5811637	5728926
6166303	6118051	6015941	5912417	5804692	5728921
6166296	6114614	6013859	5910634	5804691	5728920
6162968	6114613	6005172	5910633	5792915	5728919
6162964	6114612	6005168	5908976	5792912	5728558
6160211	6114607	5998711	5907086	5792911	5723723
6160209	6111173	5990393	5905189	5792910	5723722

5717129	5541352	5159133	6495738	5451514
5706189	5534661	5159132	6410828
5689036	5530184	5157208	6384207
5689034	5527986	5157206	6331664
5675066	5506368	5097096	6323395
5639946	5506387	5097095	6166302
5638637	5502272	5097093	6048838
5633427	5495069	5097092	7799906
5625133	5495066	5095174	7723584
5625132	5495065	5082992	7709697
5625130	5491295	5082991	7674951
5625129	5491290	5049503	7663023
5618987	5491289	4996049	7598430
5608140	5491287	4812600	7288409
5608139	5491286	4812599	7232941
5608138	5478369	4806669	7148406
5602318	5476999	480Θ652	7135618
5602317	5463173	4737596	7087426
5602312	5461171	4731499	7071384
5585538	5453564	7820883	7071376
5585537	5444178	7795395	7049485
5583210	5436390	7728195	7012172
5576472	5434346	7723582	6906239
5574209	5432068	7723581	6855864
5574206	5426041	7723580	6831208
5569822	5416254	7723579	6703539
5569821	5387758	7723578	6653528
5569619	5387755	7723577	6635805
5569816	5387754	7718852	6610908
5569817	5367109	7709707	6489538
5569816	5365014	7709623	6479732
5587861	5356799	7671253	6476291
5563326	5354941	7667100	6455762
5563325	5349119	7635798	6441272
5563323	5347081	7566818	6252135
5563322	5347080	7456340	6242381
5563321	5347079	7411117	6211432
5563320	5316930	7371935	6204434
5563055	5304720	7355100	6066780
5557038	5304719	7348473	6015943
5557035	5285004	7332657	5981837
5557034	5276265	7179965	5959178
5545814	5276264	7071386	5952486
5545813	5260503	7071385	5922928
5545812	5245125	6791013	5886243
5545809	5220114	6753460	5850020
5543575	5159134	6753459	5846333

20100071092	20100293639	20100275304
20100058498	20100293638	20100275303
20100058496	20100293637	20100275301
20090119796	20100293634	20100275300
20090106862	20100293633	20100275299
20090044294	20100293632	20100275298
20090019605	20100293630	20100275297
20080313774	20100293629	20100275296
20080235820	20100287653	20100275295
20080213871	20100287652	20100275294
20080118954	20100287651	20100275293
20080058510	20100287650	20100275292
20060206964	20100287649	20100275291
20060130183	20100287848	20100275290
20060101535	20100287647	20100275289
20050091707	20100287646	20100275288
20050076403	20100287645	20100275287
20040049802	20100287644	20100275286
20030163839	20100287643	20100273987
20030131373	20100287642	20100272880
20020138870	20100285202	20100269224
20020078477	20100281564	20100269211
20020078474	20100281563	20100269210
20100293661	20100281562	20100269209
20100293660	20100281561	20100269208
20100293659	20100281560	20100269207
20100293658	20100281559	20100269206
20100293657	20100281558	20100269205
20100293858	20100281557	20100269204
20100293655	20100278996	20100269203
20100293654	20100275332	20100269202
20100293653	20100275322	20100269201
20100293852	20100275318	20100269200
20100293651	20100275317	20100269199
20100293650	20100275316	20100269198
20100293649	20100275315	20100269197
20100293648	20100275314	20100269196
20100293647	20100275313	20100269195
20100293646	20100275312	20100263083
20100293645	20100275310	20100263082
20100293644	20100275309	20100263081
20100293643	20100275308	20100263080
20100293642	20100275307	20100263079
20100293640	20100275305	20100263078

20100263077 20100263076 20100263075 20100263074 20100263073 20100263072 20100263071 20100263070 20100263069 20100263066 20100260921 20100260920 20100257630 20100257629 20100257628 20100257627 20100257626 20100257625 20100257621 20100255175 20100251412 20100251411 20100251410 20100251406 20100251407 20100251406 20100251405 20100251403 20100251402 20100251401 20100251400 20100251399 20100251398 20100249389 20100248963 20100247733 20100242132 20100242130 20100242129 20100218269 20100196580 20100192245 20100173061 20100168455 20100146656 20100138953 20100115652

20100115649 20100115648 20100115647 20100115646 20100115645 20100115644 20100115643 20100112182 20100107272 20100107271 20100107270 20100107268 20100107267 20100100980 20100080887 20100064394 20100043094 20100043093 20100043091 20100043090 20100043088 20100043087 20100043086 20100037339 20100037336 20100037337 20100037336 20100037333 20100024064 20100024063 20100024062 20100024054 20100024052 20090288216 20090288215 20090288213 20090288212 20090288211 20090288210 20090288208 20090288207 20090288206 20090288205 20090288203 20090288202 20090288201 20090288200

20090288198 20090288197 20090288195 20090288194 20090288191 20090288189 20090288188 20090282575 20090282574 20090282573 20090282572 20090282571 20090282570 20090282569 20090282568 20090282567 20090282566 20090282565 20090282564 20090282563 20090282562 20090282561 20090282560 20090282559 20090282558 20090282557 20090282556 20090282555 20090282554 20090282553 20090282552 20090282551 20090282550 20090282549 20090282548 20090282547 20090282546 20090282545 20090282544 20090282543 20090282542 20090282541 20090282540 20090282539 20090282538 20090282536 20090282535

20090282534	20090276895	20090081353
20090282533	20090276894	20090077694
20090282532	20090276893	20090070902
20090282531	20090276892	20090070891
20090282530	20090276891	20090055957
20090282529	20090276885	20090055956
20090282528	20090276884	20090055955
20090282527	20090276883	20090031438
20090282528	20090276882	20090029861
20090282525	20090276881	20090019604
20090282523	20090276880	20090019603
20090282522	20090276879	20090019595
20090282521	20090276878	20090019594
20090282520	20090276871	20090019593
20090282519	20090276870	20090019592
20090202517	20090276869	20090019591
20090282516	20090275741	20090019590
20090282515	20090246350	20090019589
20090282514	20090241213	20090019588
20090282513	20090241212	20090019587
20090282512	20090241211	20090019586
20090282511	20090241210	20090019585
20090282510	20090235379	20090019584
20090282509	20090235378	20090019583
20090282508	20090235377	20090019582
20090282500	20090229004	20090019581
20090282499	20090229003	20090019580
20090282498	20090229002	20090019579
20090276916	20090210961	20090019578
20090276915	20090169709	20090019577
20090276914	20090165163	20090019576
20090276913	20090165162	20090019575
20090276910	20090165161	20090019574
20090276909	20090165159	20090019573
20090276908	20090165158	20090019572
20090276907	20090151020	20090019571
20090276906	20090138989	20090019570
20090276905	20090138985	20090019569
20090278904	20090133147	20090019568
20090276903	20090133146	20090019567
20090276902	20090133145	20090019565
20090276901	20090133144	20090019564
20090276900	20090133143	20090013429
20090276899	20090133142	20090013428
20090276898	20090100536	20090013427
20090276897	20090098099	20090013426
20090276896	20090083886	20090013425

20090007290 20090007289 20090007288 20090007287 20090007288 20080282422 20080282421 20080282420 20080282419 20080282418 20080282417 20080282416 20080282415 20080282414 20080282413 20080282412 20080282411 20080282410 20080282409 20080282408 20080282407 20080282406 20080282405 20080282404 20080282403 20080282402 20080282401 20080282400 20080282399 20080282398 20080282397 20080282396 20080282395 20080282394 20080282393 20080282392 20080282389 20080282388 20080282387 20080282386 20080282385 20080282384 20080262383 20080282382 20080282381 20080282380 20080282379

20080313765 20080313764 20080313763 20080313762 20080313761 20080282378 20080282377 20080282376 20080282375 20080282374 20080282373 20080282372 20080282371 20080282370 20080282366 20080280361 20080276330 20080271197 20080271196 20080271195 20080271194 20080271193 20080271192 20080271191 20080271190 20080271189 20080271188 20080271187 20080271186 20080271185 20080271184 20080271183 20080271182 20080271181 20080271180 20080271179 20080271178 20080271177 20080271176 20080271175 20080271174 20080271173 20080271172 20080271171 20080271170 20080271168 20080283713

20080313760 20080313759 20080313758 20080313757 20080282432 20080263712 20080263711 20080263706 20080263705 20080260929 20080256669 20080235819 20080227639 20080216190 20080216189 20080178345 20080178344 20080178343 20080178342 20080178341 20080178340 20080178338 20080178337 20080178336 20080178335 20080178334 20080178333 20080178332 20080178331 20080178330 20080178329 20080178328 20080178327 20080178326 20080178322 20080178320 20080178319 20080178318 20080178317 20080172781 20080172756 20080172755 20060172754 20080168576 20080155711 20080155710 20080155708

I «

20080155707	20060162007	20040237150
20080148428	20060111254	20040237149
20080148427	20060107340	20040237148
20000148426	20060101543	20040237139
20000148425	20060070140	20040221346
20080148424	20060064777	20040221344
20080148423	20060064776	20040221343
20080148422	20060059590	20040221342
20080148421	20060059589	20040221341
20080148420	20060021081	20040221339
20080070286	20060010530	20040221335
20000066202	20060010529	20040221329
20080064868	20060010528	20040221328
20080050506	20060010527	20040210958
20080022423	20060010526	20040205062
20070266456	20060010525	20040205861
20070256190	20060010524	20040205860
20070256187	20060010523	20040205859
20070256186	20060010522	20040205857
20070256105	20060010521	20040205856
20070256184	20050209664	20040205854
20070256182	20050193440	20040205849
20070256181	20050193430	20040168228
20070256180	20050193437	20040168225
20070256179	20050193436	20040168224
20070256171	20050183155	20040168223
20070256170	20050183154	20040168222
20070256155	20050183153	20040168219
20070250957	20050155114	20040148665
20070250955	20050155106	20040148664
20070250954	20050144680	20040148663
20070250952	20050144879	20040148662
20070250951	20050144678	20040148660
20070250950	20050144677	20040148659
20070250949	20050144676	20040148650
20070250947	20050144675	20040132975
20070214516	20050144674	20040111772
20070067871	20050132437	20040111770
20070054400	20050114929	20040093637
20070037700	20050114928	20040060002
20070022494	20050079494	20040055059
20070011761	20050071900	20040055058
20060288451	20050022272	20040055057
20060288447	20050022261	20040055056
20060202915	20050005332	20040055055
20060265778	20050005321	20040055054
20060174372	20040237152	20040055051

» a

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20040055047	20100162432	20090075819
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20060162021	20100186115	20090265818
20060137043	20100186113	20090265802
20060112452	20100175150	20090264351
20060064784	20100167403	20090264290
20060037102	20100162440	20090260106
20060010514	20100162425	20090260105
20050241020	20100162424	20090235392
20040194163	20100154083	20090229018
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20100287669	20100132070	20090222954
20100287662	20100122367	20090222943
20100287656	20100115667	20090210970
20100287655	20100115662	20090205078
20100285591	20100115681	20090205067
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20100281569	20100088785	20090188008
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Claims (20)

WHAT IS CLAIMED IS:

1 o

22. The method of any of the above claims wherein the plant is a gcnetically modified alfalfa, potato corn, wheat, beet, cotton, rapeseed, rice, or sugarcane plant.

23. A product comprising sugar from a feedstock obtained at least in part from a plant that has been modified with respect to a wild type varîety of the plant.

24. A product comprising an irradiated feedstock obtained at least in part from a

1. A method of making a product comprising:

physically trcating a feedstock obtained at least in part from a plant that has been modifîed with respect to a wild type variety of the plant,

5

2. The method of claim 1 wherein the feedstock comprises lignocellulosic or cellulosic material.

3. The method of claim 1 or 2 wherein the plant has been genetically modifîed.

4. The method of any of the above claims wherein the plant comprises recombinant DNA.

5 comprises irradiation of the feedstock.

6. The method of any of the above claims wherein the plant expresses a recombinant protein.

7. The method of any of the above claims wherein the plant expresses onc or more

8. The method of claim 7 wherein the recombinant material is a polymer or a macromoleculc.

9. The method of any of the above claims further comprising obtaining from the feedstock a material selected from the group consisting of pharmaceuticals,

20 nutriceuticals, proteins, fats, vitamîns, oils, fiber, minerais, sugars, carbohydrates and alcohols.

10 15. The method of claim 12 wherein irradiatîng comprises applying a total dose of from about 5 Mrad to about 50 Mrad of radiation to the feedstock.

10. The method of any of the above claims further comprising treating the feedstock with an organism and/or enzyme to produce a product.

¹⁰ 5. The method of any of the above claims wherein the plant comprises one or more recombinant genes.

11. The method of claim 10 wherein the product comprises a sugar.

12. The method of any of the above claims wherein the physical treatment

13. The method of claim 12 further comprising utilizing the irradiated feedstock as an animal feed.

14. The method of claim 12 wherein irradiatîng is performed using one or more électron beam devices.

15 plant that has been modified with respect to a wild type varicty of the plant.

25. The product of claim 24 further comprising a microorganism and/or an enzyme.

26. The product of claim 24 or 25 further comprising a liquid medium.

27. A product comprising a physically treated cellulosic or lignocellulosic

15 com stover.

15 recombinant materials.

16. The method of any of the above claims wherein the feedstock comprises a crop residue.

17. The method of claim 16 wherein the feedstock comprises corn cobs and/or

18. The method of claim 16 wherein the feedstock comprises wheat straw.

19. The method of any of the above claims wherein the plant comprises a genetically modified com or soybean plant.

20. The method of claim 11 further comprising fermenting the sugar.

i V

21. The method of any of the above claims wherein the plant has been modified with a modification selected from the group consisting of enhancement of résistance to insects, fungal diseases, and other pests and disease-causing agents; increased tolérance to herbicides; increased drought résistance; extended température range; enhanced ⁵ tolérance (o poor soil; enhanced stabilîty or shelf-life; greater yield; larger fruit size;

stronger stalks; enhanced shatter résistance; reduced time to crop maturity; more uniform germination times; higher or modified starch production; enhanced nutrient production,; modified lignin content; enhanced cellulose, hcmiccllulosc and/or lignin dégradation;; reduced recalcitrance and enhanced phytate metabolism.

20 feedstock obtained at least in part from a plant that has been modified with respect to a wild type varicty of the plant.