MX2015003073A

MX2015003073A - Processes and systems for the production of fermentation products.

Info

Publication number: MX2015003073A
Application number: MX2015003073A
Authority: MX
Inventors: Joseph J Zaher; Stephane Francois Bazzana; Keith H Burlew; James Timothy Cronin; Brian Michael Roesch; Adam Bernfeld; Michael Charles Grady; Raymond Richard Zolandz
Original assignee: Butamax Advanced Biofuels Llc
Priority date: 2012-09-12
Filing date: 2013-09-12
Publication date: 2015-07-14
Also published as: CA2883627A1; CN104919050A; BR112015005439A8; BR112015005439A2; AU2013315520B2; AU2013315520A1; NZ705079A; EP2895612A1; WO2014043288A1; JP2018121648A; KR20150054832A; JP6653576B2; JP2015528309A; AU2017201207A1

Abstract

The present invention relates to the production of fermentation products such as alcohols including ethanol and butanol, and processes employing in situ product removal methods wherein an extractant is added to the fermentation broth.

Description

PROCESSES AND SYSTEMS FOR THE PRODUCTION OF PRODUCTS FERMENTATION FIELD OF THE INVENTION The present invention relates to the production of fermentation products, such as alcohols including ethanol and butanol, and processes that use methods of in-situ removal of the product.

BACKGROUND OF THE INVENTION With the use of fermentation as a manufacturing process, a series of chemical substances and consumer products can be produced. For example, alcohols such as ethanol and butanol have a variety of industrial and scientific applications, such as fuels, reagents and solvents. Butanol is an important industrial chemical with a wide variety of applications including use as an additive for fuels, as a chemical raw material in the plastics industry and as a food grade extractant in the food and flavor industry. The production of butanol or butanol isomers from materials such as plant derived materials could minimize the use of petrochemical products and would represent an advance in the field. In addition, the production of chemicals and fuels that use materials derived from plants or other sources Ref.254695 of biomass would provide petrochemical processes with sustainable and environmentally friendly alternatives.

To produce a particular product from materials derived from plants or other sources of biomass, techniques such as genetic engineering and metabolic engineering can be used to modify a microorganism. However, in the production of butanol, for example, some microorganisms that produce butanol in high yields also have low thresholds of butanol toxicity. The removal of butanol from fermentation as it occurs is a means of managing these low butanol toxicity thresholds. Therefore, despite the low butanol toxicity thresholds of butanol producing microorganisms, there remains a need to develop efficient methods and systems for producing butanol in high yields.

To extract butanol or other products from fermentation as they are produced, which allows the microorganism to produce a high yield of butanol, the in-product removal of the product (ISPR) can be used. English) also called extractive fermentation. One method of ISPR for extracting fermentative alcohol that has been described in the art is liquid-liquid extraction (see, for example, U.S. Patent Application Publication No.2009 / 0305370). Generally, in relation to butanol fermentation, the Fermentation broth, which includes the microorganism, is contacted with an extractant before the butanol concentration reaches, for example, a toxic level. Partitions of butanol in the extractant decrease the concentration of butanol in the fermentation broth that contains the microorganism, which limits, in this way, the exposure of the microorganism to the butanol inhibitor.

To be technically and economically viable, the liquid-liquid extraction requires the contact between the extractant and the fermentation broth for an efficient mass transfer of the alcohol to the extractant; the phase separation of the extractant from the fermentation broth (during and / or after fermentation); efficient recovery and reclosing of the extractant; and the minimum reduction of the partition coefficient of the extractant during a long-term operation. Over time, the extractant can be contaminated in each recycling, for example, by the accumulation of lipids present in the biomass used as raw material for fermentation, and this contamination can lead to a concomitant reduction of the partition coefficient of the extractant.

In addition, the presence of undissolved solids during extractive fermentation can negatively affect the efficiency of alcohol production. For example, the presence of undissolved solids can decrease the mass transfer coefficient; prevent phase separation; producing the accumulation in the extractant of the oil of the undissolved solids, which leads, over time, to the reduction in the efficiency of the extraction; reduce the rate at which the extractant drops are decoupled from the fermentation broth; produce a lower fermentation efficiency in the volume of the container; and increase the extractant loss, because it is trapped in the solids and, finally, extracted as dry distillers grains with solubles (DDGS, for its acronym in English).

Therefore, there is a continuing need for alternative extractive fermentation processes that reduce the toxic effect of fermentative alcohol, such as butanol, on the microorganism and which, moreover, can reduce the degradation of the partition coefficient of an extractant. The present invention meets the needs described in the present description and provides methods, processes and systems for the fermentative production of alcohols, such as ethanol and butanol.

BRIEF DESCRIPTION OF THE INVENTION The present invention is directed to a method for recovering a fermentation product from a fermentation broth; The method comprises providing a fermentation broth comprising a microorganism, wherein the microorganism produces a fermentation product in a termenter contacting the fermentation broth with at least one extractant; and recover the fermentation product. In some embodiments, the contacting of the fermentation broth with at least one extractant takes place in the fermenter, an external unit or both. In some modalities, the external unit is an extractor. In some embodiments, the extractor is selected from siphon, decanter, centrifuge, gravity settler, phase splitter, mixer-settler, column extractor, centrifugal extractor, stirring extractor, hydrocyclone, spray tower, and combinations thereof. In some embodiments, the extractant is selected from fatty alcohols of C7 to C22, fatty acids of C7 to C22 fatty acid esters of C7 to C22, fatty aldehydes of C7 to C22, fatty amides of C7 to C22 and mixtures of these. In some embodiments, the extractant is selected from oleyl alcohol, behenyl alcohol, cetyl alcohol, lauryl alcohol, myristyl alcohol, stearyl alcohol, oleic acid, lauric acid, linoleic acid, linolenic acid, myristic acid, stearic acid, octanoic acid, decanoic acid , undecanoic acid, methyl myristate, methyl oleate, 1-nonanol, 1-decanol, 2-undecanol, 1-nonanal, 1-undecanol, undecanal, lauric aldehyde, 2-methylundecanal, oleamide, linoleamide, palmitamide, stearylamide, -ethyl-1-hexanol, 2-hexyl-1-decanol, 2-octyl-l-dodecanol, and mixtures thereof. In some embodiments, a hydrophilic solute is added to the fermentation broth. In some embodiments, the hydrophilic solute is selected from polyhydroxy compounds, polycarboxylic acids, polyol compounds, ionic salts, and mixtures thereof. In some embodiments, the contacting of the fermentation broth with at least one extractant takes place in two or more external units. In some embodiments, the contacting of the fermentation broth with at least one extractant takes place in two or more fermenters. In some embodiments, the fermenters comprise internal components or devices to improve phase separation. In some embodiments, the internal components or devices are selected from coalescing agents, baffles, perforated plates, wells, sheet separators, cones, and combinations thereof. In some modalities, real-time measurements are used to monitor the extraction of the fermentation product. In some embodiments, the extraction of the fermentation product is monitored by real-time measurements of phase separation. In some embodiments, phase separation is monitored by measurement of the phase separation rate, extractant droplet size and / or composition of the fermentation broth. In some embodiments, phase separation is monitored by conductivity measurements, dielectric measurements, viscoelastic measurements and / or ultrasonic measurements. In some modalities, the provision of a fermentation broth which comprises a microorganism takes place in two or more thermenators. In some embodiments, the fermentation product may be an alcoholic product. In some embodiments, the alcoholic product is selected from ethanol, propanol, butanol, pentanol, hexanol and fusel alcohols. In some embodiments, the microorganism comprises a butanol biosynthetic pathway. In some embodiments, the butanol biosynthetic pathway is a biosynthetic pathway of 1-butanol, a biosynthetic pathway of 2-butanol, a biosynthetic pathway of isobutanol, or a 2-butanone pathway. In some embodiments, the microorganism is a recombinant microorganism. In some embodiments, the method further comprises the steps of providing a suspension of raw material comprising a source of fermentable carbon, undissolved solids, oil and water; separating the raw material suspension and forming three streams: i) an aqueous solution comprising fermentable carbon source, (ii) a wet cake comprising solids and (iii) oil; and adding the aqueous solution to the fermentation broth. In some embodiments, the oil is hydrolysed to form fatty acids. In some embodiments, the fermentation broth is contacted with the fatty acids. In some embodiments, the oil is hydrolyzed by an enzyme. In some embodiments, the enzyme is one or more lipases or phospholipases. In some modalities, the suspension of raw material is generated by hydrolysis of the raw material. In some embodiments, the raw material is selected from rye, wheat, corn, sugar cane, barley, cellulose or lignocellulosic material, and combinations thereof. In some embodiments, the suspension of raw material is separated by means of centrifugation in bowl decanter, three-phase centrifugation, disk-disk centrifugation, centrifugation filtration, decanter centrifugation, filtration, membrane filtration, microfiltration, vacuum filtration. , band filter, pressure filtration, filtration through the use of a screen, screen separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, vortex separator, or combination of these. In some modalities, the separation of the raw material is a one-stage process. In some embodiments, the wet cake is combined with the aqueous solution. In some embodiments, the method further comprises contacting the aqueous solution with a catalyst and converting the oil from the aqueous solution into fatty acids. In some embodiments, the aqueous solution and the fatty acids are added to the fermentation broth. In some embodiments, the catalyst is deactivated.

The present invention is also directed to a system comprising one or more fermenters; they include: an entrance to receive the suspension of matter cousin; and an outlet for discharging the fermentation broth comprising the fermentation product; and one or more extractors comprising: a first entry to receive the fermentation broth; a second entry to receive the extractant; a first outlet to discharge an impoverished fermentation broth; and a second outlet to discharge an enriched extractant. In some embodiments, the system also comprises one or more liquefaction units; one or more separation means; and, optionally, one or more washing systems. In some embodiments, the separation media are selected from centrifugation in bowl decanter, three-phase centrifugation, disk-stack centrifugation, filtration centrifugation, decanter centrifugation, filtration, vacuum filtration, band filter, pressure filtration, membrane filtration, microfiltration, filtration through the use of a screen, screen separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, vortex separator, and combinations of these. In some embodiments, the system also includes online measuring devices. In some embodiments, in-line measurement devices are selected from particle size analyzers, infrared Fourier transform spectroscopes, near-infrared spectroscopes, Raman spectroscopes, high pressure liquid chromatography, viscometers, densitometers, tensiometers, droplet size analyzers, pH meters, probes for dissolved oxygen or combinations of these.

BRIEF DESCRIPTION OF THE FIGURES The accompanying figures, which are incorporated in the present description and form part of the description, illustrate the present invention and, together with the description, further explain the principles of the invention and allow a relevant expert in the art to prepare and use the invention.

Figure 1 illustrates schematically an illustrative process and system of the present invention, in which undissolved solids are removed by separation after liquefaction and before fermentation.

Figure 2 illustrates schematically an illustrative process and system of the present invention, in which an ISPR is performed downstream of the fermentation.

Figure 3 illustrates schematically another alternative process and system illustrative of the present invention, in which an oil stream is discharged.

Figure 4 schematically illustrates another alternative process and systems illustrative of the present invention, in which the wet cake is subjected to wash cycles.

Figure 5 illustrates schematically another alternative process and system illustrative of the present invention, in which an oil stream is discharged and the wet cake is subjected to washing cycles.

Figures 6A and 6B illustrate schematically another alternative process and system illustrative of the present invention, in which the aqueous solution and the wet cake combine and lead to fermentation (Figure 6A) and the aqueous solution, the oil and the wet cake combine and conduct until fermentation (Figure 6B).

Figures 7A-7D illustrate schematically alternative processes and systems illustrative of the present invention, in which the aqueous solution is subjected to conversion (eg, hydrolysis, transesterification) and / or deactivation.

Figure 8 schematically illustrates an illustrative fermentation process of the present invention that includes downstream processing.

Figure 9 schematically illustrates an illustrative fermentation process of the present invention that includes downstream processing.

Figures 10A-10M illustrate various systems that can be used in the processes described in the present description.

Figures 11A and 11B schematically illustrate multiple-pass extractant flow systems.

Figure 12 illustrates schematically an illustrative fermentation process of the present invention using in-line, in-line, line-of-line and / or in-line measurements.

Real time to monitor the fermentation processes.

Figures 13A and 13B illustrate schematically illustrative processes of the present invention to mitigate the formation of an emulsion layer.

Figure 14 illustrates schematically an illustrative process of the present invention, including fermentation, extraction and distillation processes.

Figure 15 shows the effects of the proportions between fermentation broth and extractant (aq / org) on the efficiency of the extraction column.

Figures 16A and 16B show the effects of ISPR using an external extraction column in isobutanol concentrations and glucose profiles.

Figure 17 shows the effects of ISPR using a mixer-settler at isobutanol removal rates.

Figure 18 shows the FTIR spectra of the range of starch concentrations by using in-line measurements.

Figure 19 shows the FTIR spectra of the starch concentration of the wet cake during processing of corn temper.

Figure 20 shows the FTIR spectra of corn oil during processing of corn temper.

Figure 21 shows a real-time measurement of isobutanol in COFA.

DETAILED DESCRIPTION OF THE INVENTION Unless defined otherwise, all scientific and technical terms used herein have the same meaning as commonly understood by a person with ordinary knowledge in the subject matter to which this invention pertains. In case of dispute, the present application will prevail along with the relevant definitions. In addition, unless otherwise required, terms in the singular will include pluralities and terms in the plural will include the singular. All publications, patents and other references mentioned in the present description are incorporated by reference in their entirety for all purposes.

To further define this invention, the following terms and definitions are provided in the present description.

As used in the present description, the terms "comprising", "comprising", "including", "including", "having", "having", "containing" or "containing" or another variation of these implies the inclusion of an integer or group of integers mentioned, but not the exclusion of any other integer or group of integers. For example, a composition, a mixture, a process, a method, an article or an apparatus comprising a list of elements is not necessarily limited only to those elements, but may include others that are not expressly listed or are inherent to such composition, mixture, process, method, article or apparatus. In addition, unless specifically stated otherwise, the disjunction is related to an "or" inclusive and not an "or" excluding. For example, a condition A or B is satisfied by any of the following criteria: A is true (or current) and B is false (or not current), A is false (or not current) and B is true (or current) , and both A and B are true (or current).

In addition, the indefinite articles "a" and "ones" that precede an element or component of the invention are intended to be non-restrictive with respect to the number of instances, i.e. occurrences of the element or component Therefore, "a" or "ones" must be construed to include one or at least one, and the singular form of the word of the element or component includes, in addition, the plural, unless the The number obviously indicates that it is singular.

As used in the present description, the term "invention" or "present invention" is a non-limiting term that is not intended to refer to a single embodiment of the invention, but encompasses all possible embodiments described in the application.

As used in the present description, the term "about", which modifies the amount of an ingredient or reagent used in the invention, refers to the variation which can be produced in numerical quantity, for example, through liquid handling procedures and typical measurements used to prepare concentrates or solutions in the real world; through inadvertent errors in these procedures; through differences in the manufacture, provenance or purity of the ingredients used to prepare the compositions or carry out the methods; and similar. The term "approximately" also encompasses amounts that differ due to different equilibrium conditions for a composition resulting from a particular initial mixture. Whether or not modified by the term "approximately", the claims include equivalents for the quantities. In one embodiment, the term "approximately" means an amount within 10% of the numerical value reported, alternatively, within 5% of the numerical value reported.

As used herein, "biomass" refers to a natural product that contains hydrolysable polysaccharides that provide fermentative sugars and / or starches, which include any sugar and starch derived from natural resources, such as corn, sugar cane, wheat. , cellulose or lignocellulosic material and materials comprising cellulose, hemicellulose, lignin, starch, oligosaccharides, disaccharides and / or monosaccharides, and mixtures thereof. The biomass may further comprise additional components, such as proteins and / or lipids. The biomass it may be derived from a single source or it may comprise a mixture derived from more than one source, for example, the biomass may comprise a mixture of maize ears and corn stubble, or a mixture of grass and leaves. Biomass includes, but is not limited to, bioenergy crops, agricultural residues, municipal solid waste, industrial solid waste, papermaking sediments, garden waste, and forestry and forestry waste (for example, forest thinning). Examples of biomass include, but are not limited to, corn, corncobs, crop residues, such as corn husks, corn stover, pasture, wheat, rye, wheat straw, spelled, triticale, barley, straw. barley, oats, hay, rice, rice straw, grass rod, potato, sweet potato, cassava, Jerusalem artichoke, sugarcane bagasse, sorghum, sugar cane, sugar beet, fodder beet, soybeans, palm, coconut, seed rapeseed, safflower, sunflower, millet, eucalyptus, miscanthus, components obtained from the grinding of grains, trees (for example, branches, roots, leaves), wood shavings, sawdust, shrubs and bushes, vegetables, fruits, flowers, manure , and mixtures of these. For example, temper, juice, molasses or hydrolyzate can be formed from the biomass by any process known in the art to process the biomass for fermentation purposes, such as milling and liquefaction.

For example, to obtain a hydrolyzate containing fermentable sugars, cellulose and / or lignocellulosic biomass can be processed by any method known to a person skilled in the art, such as pretreatment with low levels of ammonia described in the patent application publication of I know. UU no. 2007/0031918, which is incorporated in the present description as a reference. Typically, the enzymatic saccharification of cellulosic and / or lignocellulosic biomass uses a set of enzymes (eg, cellulases, xylanases, glycosidases, glucanases, lyases) to break down cellulose and hemicellulose to produce a hydrolyzate containing sugars, including glucose, xylose and arabinose. Saccharification enzymes suitable for cellulosic and / or lignocellulosic biomass are described in Lynd, et al. (Microbiol.Mol. Biol. Rev. 66: 506-577, 2002).

"Fermentable carbon source" or "fermentable carbon substrate", as used in the present description, refers to a carbon source that can be metabolized by microorganisms. Suitable fermentable carbon sources include, but are not limited to, monosaccharides, such as glucose or fructose; disaccharides, such as lactose or sucrose; oligosaccharides; polysaccharides, such as starch or cellulose; a carbon substrate; and mixtures of these.

As used in the present description, "sugar "fermentable" refers to one or more sugars that the microorganisms described in the present disclosure can metabolize to produce fermentation products.

As used in the present description "raw material" refers to a feed in a fermentation process; the feed contains a fermentable carbon source with or without undissolved solids, and oil, and where appropriate, the feed contains the fermentable carbon source before or after the fermentable carbon source has been removed from the starch or obtained from the decomposition of complex sugars by further processing, such as liquefaction, saccharification or other process. The raw material includes or can be derived from biomass. Suitable raw materials include, but are not limited to, rye, wheat, corn, corn temper, cane, cane temper, barley, cellulosic material, lignocellulosic material, or mixtures thereof. When reference is made to "raw material oil" it is understood that the term covers the oil produced from a certain raw material.

As used in the present description, "fermentation broth" refers to a mixture of water, fermentable carbon sources (e.g., sugars), dissolved solids, optionally, microorganisms that produce fermentation products (e.g., alcoholic product) , optionally, fermentation products (for example, alcoholic product), and other components. In some embodiments, fermentation broth refers to the material maintained in the fermenter in which the fermentation product (eg, alcohol product) is produced by the metabolism of fermentable carbon sources by the microorganisms. As used in the present description, the term "fermentation broth" can sometimes be used synonymously with "fermentation medium" or "fermented mixture". In some embodiments, the fermentation broth comprising the alcoholic product may be referred to as fermentation beer or beer.

As used herein, "fermenting" or "fermentation vessel" refers to the unit in which the fermentation reaction is carried out, whereby the fermentation product (eg, alcohol product, such as ethanol) is produced. or butanol) from fermentable carbon sources. The term "fermenter" can be used in the present description as a synonym for "fermentation vessel".

As used in the present description, "liquefaction unit" refers to the unit in which liquefaction is effected. Liquification is the process in which oligosaccharides are released from the raw material. In some embodiments in which the raw material is corn, the oligosaccharides are released from the corn starch content during liquefaction.

As used in the present description, "saccharification unit" refers to the unit in which saccharification is effected (i.e., the decomposition of oligosaccharides into monosaccharides). When the fermentation and saccharification take place simultaneously, the saccharification unit and the fermentor can be the same unit.

"Sugar", as used in the present description, refers to oligosaccharides, disaccharides, monosaccharides and / or mixtures thereof. The term "saccharide" also includes carbohydrates including starches, dextrans, glycogens, cellulose, pentosans, as well as sugars.

As used herein, "saccharification enzyme" refers to one or more enzymes capable of hydrolyzing polysaccharides and / or oligosaccharides, eg, alpha-1,4-glycosidic linkages of glycogen or starch. Saccharification enzymes can include enzymes capable of hydrolyzing cellulosic and also lignocellulosic materials.

As used in the present description, "undissolved solids" refers to non-fermentable portions of raw material, eg, germ, fiber, gluten and any additional component that does not dissolve in aqueous media. For example, the non-fermentable portions of raw material include the portion of raw material that is held as a solid and can absorb liquid from the fermentation broth.

"Oil", as used in the present description, is refers to lipids obtained from plants (for example, biomass) or animals. Examples of oils include, but are not limited to, tallow, corn, cañola, capric / caprylic triglycerides, castor bean, coconut, cottonseed, fish, jojoba, lardo, linseed, bucy leg oil, ocytic oil, palm , peanut, rapeseed, rice, safflower, soy, sunflower, tung, jatropha, and mixtures of vegetable oils.

"Alcoholic product", as used in the present description, refers to any alcohol that can be produced by means of a microorganism in a fermentation process that uses biomass as a source of fermentable carbon substrate. Alcohol products include, but are not limited to, alkyl alcohols from Ci to Cs. In some embodiments, the alcohol products are alkyl alcohols from C2 to Ce. In other embodiments, the alcohol products are alkyl alcohols from C2 to C5. It will be appreciated that the alkyl alcohols of Ci to Cs include, but are not limited to, methanol, ethanol, propanol, butanol, pentanol and hexanol. Similarly, alkyl alcohols from C2 to Cs include, but are not limited to, ethanol, propanol, butanol, pentanol and hexanol. In some embodiments, the alcoholic product may also include fusel alcohols (or fusel oils) and glycerol. "Alcohol" is also used in the present description with reference to an alcoholic product.

As used in the present description, "butanol" is refers to the isomers of butanol: 1-butanol (1-BuOH), 2-butanol (2-BuOH), tert-butanol (t-BuOH) and / or isobutanol (iBuOH, i-BuOH, I-BUOH, known iB , in addition, as 2-methyl-1-propanol) either individually or as mixtures thereof. Conveniently, when referring to butanol esters, the terms "butyl esters" and "butanol esters" may be used interchangeably.

"Propanol", as used in the present description, refers to the isomers of propanol isopropanol or 1-propanol.

As used in the present description, "pentanol" refers to the isomers of pentanol: 1-pentanol, 3-methyl-1-butanol, 2-methyl-1-butanol, 2,2-dimethyl-1-propanol, pentanol, 2-pentanol, 3-methyl-2-butanol or 2-methyl-2-butanol.

As used in the present description, "product in-product removal (ISPR)" refers to the selective removal of a specific product from a biological process, such as fermentation, to control the concentration of the product in the custom biological process. that the product is generated.

As used in the present description, "extractant" refers to a solvent used to extract a fermentation product (e.g., alcoholic product). Occasionally, as used in the present description, the term "extractant" can be used as a synonym of "solvent" As used in the present description, "water immiscible" refers to a chemical component, such as a extractant or solvent, which is not capable of mixing with an aqueous solution, such as fermentation broth, to form a liquid phase.

"Carboxylic acid", as used in the present description, refers to any organic compound with the general chemical formula -C00H, wherein a carbon atom is attached to an oxygen atom by means of a double bond to form a carbonyl group (-C = 0) and a hydroxyl group (-0H) by means of a single bond. A carboxylic acid may be in the form of the protonated carboxylic acid, in the form of a salt of a carboxylic acid (for example, an ammonium, sodium or potassium salt) or as a mixture of protonated carboxylic acid and a salt of a carboxylic acid . The term carboxylic acid may describe a single species of chemical substance (eg, oleic acid) or a mixture of carboxylic acids such as may be produced, for example, by the hydrolysis of fatty acid esters derived from biomass or triglycerides, diglycerides, monoglycerides and phospholipids.

"Fatty acid", as used in the present description, refers to a carboxylic acid (e.g., aliphatic monocarboxylic acid) having carbon atoms of C4 to C28 (mostly, carbon atoms of C12 to C24), which can be saturated or unsaturated. The fatty acids may also be branched or unbranched. The fatty acids can be derived from, or be contained in esterified form in, a animal or vegetable fat, oil or wax. Fatty acids can exist naturally in the form of glycerides in fats and fatty oils or can be obtained by hydrolysis of fats or by synthesis. The term fatty acid can describe a single chemical species or a mixture of fatty acids. In addition, the term fatty acid encompasses free fatty acids.

"Fatty alcohol", as used in the present description, refers to an alcohol having an aliphatic chain of 4 to 22 carbon atoms (C4 to C22), saturated or unsaturated.

"Fatty aldehyde", as used in the present description, refers to an aldehyde having an aliphatic chain of 4 to 22 carbon atoms (C4 to C22), saturated or unsaturated.

As used in the present description, "fatty amide" refers to an amide having a saturated or unsaturated aliphatic chain of C4 to C22 carbon atoms.

As used in the present description, "fatty ester" refers to an ester having a saturated or unsaturated aliphatic chain of C4 to C22 carbon atoms.

As used in the present description, "aqueous phase" refers to the aqueous phase of, for example, a biphasic mixture containing, for example, a liquid phase and a vapor phase; to the aqueous phase of a three phase mixture containing two liquid phases (e.g., an organic phase and an aqueous phase) and a vapor phase; to the aqueous phase of a biphasic or triphasic mixture, wherein the aqueous phase contains a a certain amount of solids in suspension, or a quadriplegic mixture, comprising a vapor phase, an organic phase, an aqueous phase and a solid phase. In some embodiments, a three phase mixture may comprise a vapor phase, a liquid phase and a solid phase. In some embodiments, an aqueous phase can be obtained by contacting a fermentation broth with an organic immiscible water extractant. In one embodiment of a process described in the present description that includes fermentative extraction, the term "fermentation broth" can refer to the aqueous phase in the fermentative extraction of two phases.

As used in the present description, "organic phase" refers to the non-aqueous phase of a mixture (eg, biphasic mixture, three-phase mixture, quadriplegic mixture) obtained by contacting a fermentation broth with an organic extractant immiscible with Water. As used in the present description, the term "organic phase" can sometimes be used as a synonym for "extractant phase".

As used in the present description, "effective title" refers to the total amount of a particular fermentation product (eg, alcohol product) produced by fermentation, per liter of fermentation broth.

"Portion", as used in the present description with reference to a process stream, refers to any fraction of the stream that retains the composition of the current, which includes the complete current, as well as any component or components of the current, which include all the components of the current.

The present invention provides processes and methods for producing fermentation products, such as alcoholic products, with the use of fermentation. Other fermentation products that can be produced with the use of the processes and methods described in the present disclosure include propanediol, butanediol, acetone, acids such as lactic acid, acetic acid, butyric acid, and propionic acid; gases such as methane hydrogen and carbon dioxide; amino acids; vitamins such as biotin, vitamin B2 (riboflavin), vitamin B12 (for example, cobalamin), ascorbic acid (for example, vitamin C), vitamin E (for example, α-tocopherol) and vitamin K (for example, menaquinone); antibiotics such as erythromycin, penicillin, streptomycin and tetracycline; and other products such as citric acid, invertase, sorbitol, pectinase and xylitol.

The present invention provides processes and systems for producing an alcoholic product by fermentative processes and recovering an alcohol product produced by a fermentative process. As an example of one embodiment of the processes described in the present description, fermentation can be initiated by the direct introduction of raw material into a fermenter. In some modalities, one or more can be used fermenters in the processes described in the present description. Suitable raw materials include, but are not limited to, rye, wheat, corn, corn temper, cane, cane temper, barley, cellulosic material, lignocellulosic material or mixtures thereof. These raw materials can be processed through the use of methods such as dry milling or wet milling. In some embodiments, prior to introduction into the triturator, the raw material may be liquefied to create a suspension of raw material which may comprise undissolved solids, a ferrous carbon source (e.g., sugar) and oil. The liquefaction of the raw material can be achieved by any known liquefaction process, which includes, but is not limited to, acid process, enzymatic process (e.g., alpha-amylase), acid-enzymatic process, or combinations thereof. In some embodiments, liquefaction may take place in a liquefaction unit.

If the suspension of raw material is introduced directly into the fermenter, undissolved solids and / or oil can interfere with the efficient removal and recovery of an alcoholic product. Particularly, when liquid-liquid extraction is used to extract an alcoholic product from the fermentation broth, the presence of undissolved solids (eg, particulates) can cause system inefficiencies, including, but not limited to, slowing down mass transfer of the product alcohol to the extractant by interfering with the contact between the extractant and the fermentation broth; create or promote an emulsion in the fermenter and, in that way, interfere with the phase separation of the extractant and the fermentation broth; reduce the efficiency of recovery and recycling of the extractant, because at least a portion of extractant and alcoholic product is "trapped" in the solids that can be removed as dry distillers grains with solubles (DDGS); reduce the volumetric efficiency of the fermentor, because there are solids that occupy volume in the fermenter and because the extractant is decoupled from the fermentation broth with greater slowness; and shorten the extractant life cycle due to oil contamination. These effects can translate into a higher capital and operating cost. In addition, the extractant "trapped" in DDGS can detract value and qualify to market DDGS as animal feed. Therefore, to avoid and / or minimize these problems, at least a portion of the undissolved solids can be extracted from the raw material suspension before the addition of the raw material suspension to the fermenter. The extraction activity and the efficiency of the production of the alcoholic product can be increased when the extraction is carried out in a fermentation broth from which the undissolved solids were removed.

In the present description, the processes and systems to process raw material and generate a suspension of raw material, and to separate the raw material suspension and generate an aqueous phase comprising a fermentable carbon source and a solid phase (eg, wet cake) are described with reference to the figures. As shown in Figure 1, in some embodiments, the system includes liquefaction 10 configured to liquefy raw material to create a suspension of raw material. For example, the raw material 12 can be introduced in the liquification 10 (for example, by means of an input in the liquefaction unit). The raw material 12 can be any suitable biomass material known in the art, including, but not limited to, barley, oats, rye, sorghum, wheat, triticale, spelled, millet, sugar cane, corn or combinations thereof. those containing a fermentable carbon source, such as sugar and / or starch. In addition, in the liquefying 10 water can be introduced.

The process of liquefaction of the raw material 12 involves the hydrolysis of the starch from the raw material 12 to water soluble sugars. In addition to the liquefaction unit, any known liquefaction process used in the industry may be used, including, but not limited to, an acid process, an enzymatic process or an acid-enzymatic process. Such processes can be used alone or in combination. In some embodiments, the enzymatic process can be used by introducing a suitable enzyme 14, for example, alpha-amylase, to the liquation 10. Examples of alpha-amylases that can be used in the processes and systems of the present invention are described in US Pat. UU no. 7,541,026; US patent application publication UU no. 2009/0209026; US patent application publication UU No..2009 / 0238923; US patent application publication UU no. 2009/0252828; US patent application publication UU no. 2009/0314286; US patent application publication UU no. 2010/02278970; US patent application publication UU no. 2010/0048446; US patent application publication UU No. 2010/0021587, the content of which is incorporated in its entirety in the present description as a reference.

In some embodiments, the enzymes for liquefaction and / or saccharification can be produced by the microorganism. Examples of microorganisms that produce such enzymes are described in U.S. Pat. UU no. 7,498,159; US patent application publication UU no. 2012/0003701; US patent application publication UU no. 2012/0129229; PCT international patent publication no. WO 2010/096562; and PCT international publication no. WO 2011/153516, the complete contents of which are incorporated herein by reference. In some embodiments, the enzymes for liquefaction and / or saccharification can be expressed by a microorganism that also produces an alcoholic product. In some embodiments, enzymes for liquefaction and / or saccharification can be expressed by a microorganism that also expresses a butanol biosynthetic pathway. In some embodiments, the butanol biosynthetic pathway may be the biosynthetic pathway of 1-butanol, 2-butanol biosynthetic pathway, isobutanol biosynthetic pathway, or 2-butanone biosynthetic pathway.

The process of liquefaction of the raw material 12 creates the suspension of raw material 16 (also known as a temper or thick temper) which includes the fermentable carbon source (for example, sugar) and undissolved solids. In some embodiments, the suspension of raw material 16 may include the fermentable carbon source (e.g., sugar), oil and undissolved solids. The undissolved solids can be non-fermentable portions of the raw material 12. In some embodiments, the raw material 12 can be corn, such as unfractionated and dry-ground corn grains, and the raw material suspension 16 is a suspension of corn templa. The raw material suspension 16 can be discharged from an outlet of the liquefaction 10 and can be conducted to the separation 20.

The partition 20 has an inlet to receive the suspension of raw material 16 and can be configured to remove the undissolved solids from the suspension of matter premium 16. Separation 20 may also be configured to remove oil, and / or oil and undissolved solids. The separation 20 may agitate or rotate the suspension of raw material 16 to create a liquid phase or aqueous solution 22 and a solid phase or wet cake 24.

The aqueous solution 22 may include sugar, for example, in the form of oligosaccharides, and water. The aqueous solution 22 may comprise at least about 10% by weight of oligosaccharides, at least about 20% by weight of oligosaccharides or at least about 30% by weight of oligosaccharides. The aqueous solution 22 can be discharged from the separation 20 by means of an outlet. In some embodiments, the exit may be located in the vicinity of the upper part of the partition 20.

The wet cake 24 may include undissolved solids. The wet cake 24 can be discharged from the partition 20 by means of an outlet. In some embodiments, the outlet may be located in the vicinity of the bottom of the separation 20. The wet cake 24 may also include a portion of sugar and water. The wet cake 24 may be washed with more water in the separation 20, after the aqueous solution 22 has been discharged from the separation 20. Alternatively, the wet cake 24 may be washed with more water by additional separation devices. Washing the wet cake 24 recovers the sugar (for example, oligosaccharides) present in the wet cake, and the recovered sugar and water can be recielated to the liquefaction 10. After washing, the wet cake 24 can be further processed by any known suitable process to form dry distillers grains with solubles. (DDGS). The formation of DDGS from the wet cake 24 formed in the separation 20 has several advantages. Because the undissolved solids do not go to the heater, the DDGS are not subject to the conditions of the termendor. For example, DDGS does not come into contact with the microorganisms present in the thermenter or any other substance that may be present in the thermenter (for example, extractant and / or alcoholic product) and, therefore, the microorganism and / or other substances do not get trapped in DDGS. These effects provide advantages for the further processing and commercialization of DDGS, for example, as animal feed.

The separation 20 can be any conventional separation device used in the industry, including, for example, centrifuges, such as a bowl decanter centrifuge, three-stage centrifuge, disk-stack centrifuge, filter centrifuge or decanter centrifuge. In some embodiments, the removal of the undissolved solids from the suspension of raw material 16 can be effected by filtration, vacuum filtration, filter band, pressure filtration, membrane filtration, microfiltration, filtration through the use of a screen, sieve separation, gratings or grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, settling gravity, vortex separator or any other method or device that can be used to separate solids from liquids. In some embodiments, separation 20 is a one-step process. In one embodiment, undissolved solids can be removed from the raw material suspension 16 to form two product streams, for example, an aqueous oligosaccharide solution containing a lower concentration of solids, compared to the suspension of raw material 16, and a wet cake containing a higher concentration of solids, as compared to the raw material suspension 16. In addition, if for example, centrifugation of three phases is used for the removal of the solids from the suspension of raw material 16, A third stream containing oil can be generated. Several streams of product can be generated by the use of different separation techniques or combinations of these.

A three-phase centrifuge can be used for the three-phase separation of the raw material suspension, such as the separation of the raw material suspension to generate two liquid phases (e.g. oily stream) and a solid phase (for example, solids or wet cake) (see, for example, Flottweg Tricanter®, Flottweg AG, Vilsibiburg, Germany). The two liquid phases can be separated and decanted, for example, from the bowl of the centrifuge by means of two discharge systems to avoid cross contamination, and the solids phase can be removed by a separate discharge system.

In some modalities that use corn as a raw material, a three-phase centrifuge can be used to simultaneously remove the solids and corn oil from the liquefied corn temper. The solids may be undissolved solids remaining after the starch has been hydrolysed to soluble oligosaccharides during liquefaction. Corn oil can be released from the corn grain germ during grinding and / or liquefaction. In some embodiments, the three-phase centrifuge may have one supply current and three output streams. The supply stream may consist of liquefied corn pulp produced during liquefaction. The temper may consist of an aqueous solution of oligosaccharides (e.g., liquefied starch); non-dissolved solids consisting of insoluble components of corn other than starch; and corn oil consisting of glycerides and free fatty acids. The three outflow currents of the three-phase centrifuge can be a wet cake containing most of the undissolved solids of the templa; a heavy centering stream containing most of the liquefied starch in the templa; and a light centering current that contains most of the corn oil in the templa. The heavy centering current can be introduced into the fermentation. To recover the soluble starch from the wet cake, it can be washed with recirculated process water, such as evaporator condensate and / or countercurrent water as described in the present description. The light centering current can be marketed as a coproduct, converted to another coproduct or used in processes such as converting corn oil to corn oil fatty acids (COFA). In some embodiments, COFAs can be used as an extractant.

With reference to Figure 1, the fermentation 30 (or fermentor 30), configured to ferment the aqueous solution 22 to produce an alcoholic product, has an inlet to receive the aqueous solution 22. The fermentation 30 can be any suitable fermentor known in the art. matter. Fermentation 30 may include fermentation broth. In some embodiments simultaneous saccharification and fermentation (SSF) may occur within fermentation 30. Any known saccharification process that is used in the industry and that includes, but is not limited to, a process may be used. acid, an enzymatic process or an acid-enzymatic process. In some embodiments, an enzyme 38 (for example, such as glucoamylase) can be introduced into an entrance of the fermentation 30 to hydrolyse the oligosaccharides of the aqueous solution 22 and form monosaccharides. Examples of glucoamylases that can be used in the processes and systems of the present invention are described in US Pat. UU no. 7,413,887; US patent UU no. 7,723,079; US patent ication publication UU no. 2009/0275080; US patent ication publication UU No. 2010/0267114; US patent ication publication UU No. 2011/0014681; and the publication of US patent ication. UU No. 2011/0020899, the content of which is incorporated in its entirety in the present description as a reference. In some embodiments, glucoamylase can be expressed by the microorganism. In some embodiments, glucoamylase can be expressed by a microorganism that also produces an alcoholic product. In some embodiments, glucoamylase can be expressed by a microorganism that also expresses a butanol biosynthetic pathway. In some embodiments, the butanol biosynthetic pathway may be the biosynthetic pathway of 1-butanol, 2-butanol biosynthetic pathway, isobutanol biosynthetic pathway, or 2-butanone biosynthetic pathway.

In some embodiments, enzymes, such as glucoamylases, may be added to the liquefaction. The adition of Enzymes, such as glucoamylases, upon liquefaction can reduce the viscosity of the raw material suspension or the liquefied temper and can improve the separation efficiency. In some embodiments, any enzyme capable of reducing the viscosity of the raw material suspension (for example, Viscozyme®, Sigma-Aldrich, St. Louis, MO) can be used. The viscosity of the raw material can be measured by any method known in the art (for example, viscometers, rheometers).

The microorganism 32 can be introduced into the fermentation 30. In some embodiments, the microorganism 32 can be included in the fermentation broth. In some embodiments, the microorganism 32 can be propagated in a separate container or tank (e.g., the propagation tank). In some embodiments, microorganisms from the propagation tank can be used to inoculate one or more fermentors. In some embodiments, one or more propagation tanks may be used in the processes and systems described in the present disclosure. In some embodiments, the propagation tank may be from about 2% to about 5% the size of the thermistor. In some embodiments, the propagation tank may comprise one or more of the following: temper, water, enzymes, nutrients, extractant and microorganisms. In some embodiments, the alcoholic product can be produced in the propagation tank.

In some embodiments, the microorganism 32 may be bacteria, cyanobacteria, filamentous fungi or yeast. In some embodiments, microorganism 32 metabolizes sugar from aqueous solution 22 and produces the alcoholic product. In some embodiments, the microorganism 32 can be a recombinant microorganism. In some embodiments, the microorganism 32 can be immobilized, for example, by means of adsorption, covalent attachment, cross-linking, entrapment and encapsulation. Methods for encapsulating cells are known in the art, for example, as described in the US patent ication publication. UU no. 2011/0306116, which is incorporated herein by reference.

In some embodiments, in-product removal (ISPR) can be used to remove the alcoholic product from fermentation 30 as the microorganism 32 produces the alcoholic product. In some embodiments, liquid-liquid extraction for ISPR can be used. In some embodiments, the fermentation 30 may have an inlet to receive extractant 34. In some embodiments, the extractant 34 may be added to the fermentation broth downstream of the fermentation 30. The dotted lines represent alternative means of adding the extractant 34 to the fermentation 30 or downstream of the fermentation 30. In some modalities, the ISPR can be performed in a propagation tank. In some modalities, the ISPR can be performed in the fermenter and the propagation tank. In some embodiments, the ISPR can be performed at the beginning (for example, time 0) of the fermentation and / or propagation. When starting the ISPR at the beginning of the fermentation and / or propagation, the concentration of alcoholic product in the fermenter and the propagation tank can be maintained at low levels and, therefore, minimize the effects of the alcoholic product on the microorganism and allow the microorganism reaches a greater cellular mass. In some embodiments, extractant may be added to the propagation tank. In some embodiments, extractant may be added before inoculation of the propagation tank. In some embodiments, extractant may be added after inoculation of the propagation tank. In some embodiments, extractant may be added at various time points after inoculation of the propagation tank. In some embodiments, extractant may be added to the fermenter. In some embodiments, extractant may be added before inoculation of the fermentor. In some embodiments, extractant may be added after inoculation of the fermentor. In some embodiments, extractant may be added at various time points after inoculation of the fermentor. In some modalities, you can add extractant to the fermenter and the propagation tank. In the present description, examples of liquid-liquid extraction are described. The processes for producing and recovering alcohols from a fermentation broth with the use of extractive fermentation are described in the US patent application publication. UU no. 2009/0305370; US patent application publication UU no. 2010/0221802; US patent application publication UU No. 2011/0097773; US patent application publication UU no. 2011/0312044; US patent application publication UU no. 2011/0312043; and PCT international patent publication no. WO 2011/159998; the complete content of which is incorporated herein by reference.

The extractant 34 comes into contact with the fermentation broth and forms the stream 36 comprising, for example, a biphasic mixture (for example, phase enriched in extractant with alcoholic product and exhausted aqueous phase of alcoholic product). In some embodiments, the stream 36 may be a quadriplegic mixture comprising, for example, a vapor phase, an organic phase, an aqueous phase and a solid phase. The alcoholic product, or a portion thereof, of the fermentation broth is transferred to the extractant 34. In some embodiments, the stream 36 may be discharged through a fermentation outlet 30.

The alcohol product can be separated from the extractant in stream 36 with the use of conventional techniques.

In some embodiments, internal components of the fermenter or devices may be used to improve phase separation between the fermentation broth and the extractant. For example, the internal component or device can serve as a coalescing agent to promote phase separation between the fermentation broth and the extractant, and / or act as a physical barrier to improve phase separation. These internal components of the fermenter or devices can avoid, in addition, that the solids are deposited in the phase (or layer) of the extractant, promote the coalescence of aqueous droplets that can be entrained in the extractant layer and promote the removal of exhaust gases (for example, CO2, air) and , in this way, minimize the alteration of the phase of the extractant and / or the liquid-liquid interface. Examples of internal components or devices that may be used in the processes and systems described in the present disclosure include, but are not limited to, baffle plates, perforated plates, deep wells, sheet separators, cones, and the like. In some embodiments, the perforated plate may have a flat, horizontal perforated plate. In some embodiments, the cone may be an inverted cone or one or more concentric cones. In some modalities, the components internal can be rotary. In some embodiments, the internal components or devices may be located at or above the level of the liquid-liquid interface of the fermentation broth and the extractant. In some embodiments, a coalescence filter may be added and / or the exit ports may be relocated to improve coalescence and recovery of the aqueous phase.

In some embodiments, before the ISPR and / or after the completion of the fermentation, the stream 35 may be discharged from an inlet of the fermentation 30. The discharged stream 35 may include the microorganism 32. The microorganism 32 may be separated from the stream 35, for example, by centrifugation or membrane filtration. In some embodiments, the removal of the microorganism before the addition of extractant to the fermentation broth does not expose the microorganism to the extractant and, therefore, the microorganism is not exposed to the negative impact that the extractant could have on it. In addition, removal of the microorganism upstream of the extraction process allows a more aggressive extraction process to be used to recover the alcoholic product (for example, heating or cooling the mixture to improve separation, use of a larger KD and / or an extractant). of greater selectivity, or an extractant with improved properties, but of lower biocompatibility). In some modalities, the microorganism 32 can be recielated to fermentation 30, which can increase the speed of production of the alcoholic product and, in that way, increase the efficiency in the production of alcoholic product.

With reference to Figure 2, in some embodiments, the ISPR can be made downstream of the fermentation 30. In some embodiments, the stream 33, which includes the alcoholic product and the microorganism 32, can be discharged from an outlet of the fermentation 30 and conducted downstream, for example, to an extraction column for the recovery of the alcoholic product. In some embodiments, the stream 33 can be processed by separating the microorganism 32 before the ISPR. For example, removal of microorganism 32 from stream 33 can be effected by centrifugation, filtration, vacuum filtration, band filter, pressure filtration, membrane filtration, microfiltration, filtration through the use of a screen, screen separation, grids. or separation by grid, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, vortex separator or any other separation method or device that can be used to separate solids (eg, microorganisms) from liquids. After removal of the microorganism 32, the stream 33 can be conducted to an extraction column for recovery of the alcoholic product.

Figures 3 to 6 illustrate further embodiments of the processes and systems described in the present description. Figures 3 to 6, which include the options for the addition of extractant to the triturator (for example, to generate current 36) or the downstream extraction of the triturator (for example, to generate current 33), are similar to Figures 1 and 2, respectively, and, therefore, will not be described again in detail.

With reference to Figure 3, the systems and processes of the present invention can include the discharge of the oil 26 from an outlet of the separation 20. The suspension of raw material 16 can be separated into a first liquid phase or aqueous solution 22, comprising a fermentable sugar, a solid phase or wet cake 24, which comprises undissolved solids, and a second liquid phase, which comprises the oil 26 that can come out of the separation 20. In some embodiments, the separation of the raw material suspension 16 In a first liquid phase, a second liquid phase and a solid phase can take place in a single step. In some embodiments, the raw material 12 is corn and the oil 26 is corn oil. In some embodiments, the oil 26 can be conducted to a storage tank or any suitable unit for oil storage. To discharge the aqueous solution 22, the wet cake 24 and the oil 26 can be used any device of suitable separation, for example, a three-phase centrifuge. In some embodiments, when the raw material is corn, a portion of the raw material oil 12, such as corn oil, remains in the wet cake 24. In some embodiments, when the oil 26 is removed from the raw material 12 ( for example, corn) by means of separation 20, the fermentation broth in fermentation 30 includes a reduced amount of corn oil.

As described in the present description, in some embodiments, the oil may be separated from the raw material or raw material in suspension and may be stored in an oil storage unit. For example, the oil can be separated from the raw material or suspension of raw material by the use of any suitable means of separation, which includes a three-phase centrifuge or mechanical extraction. To improve the removal of oil from the raw material or suspended raw material, oil extraction aids, such as surfactants, anti-emulsifiers or flocculants, as well as enzymes can be used. Examples of oil extraction aids include, but are not limited to, non-polymeric liquid surfactants; talcum powder; microtalco powder; salts (NaOH); calcium carbonate; and enzymes such as Pectinex® Ultra SP-L, Celluclast® and Viscozyme® L (Sigma-Aldrich, St. Louis, MO) and NZ 33095 (Novozymes, Franklinton, NC).

As illustrated in Figure 4, if the oil is not discharged separately, it can be removed with the wet cake 24. When the wet cake 24 is removed by means of the separation 20, in some embodiments, a portion of the oil from the raw material 12, such as corn oil when the raw material is corn, remains in the wet cake 24. The wet cake 24 can be conducted to mixing 60 and combined with water or other solvents to form the wet cake mixture 65. In some modalities, the water may be fresh water, countercurrent water, cooking water, process water, Lutter water, evaporation water or any available water source in the fermentation processing facility, or any combination thereof. The wet cake mixture 65 can be led to the separation 70 and produce the washing centering 75 comprising fermentable sugars recovered from the wet cake 24, and the wet cake 74. The washing centering 75 can be recielated to the liquefaction 10.

In some embodiments, separation 70 may be any separation device capable of separating solids and liquids including, for example, centrifugation in bowl decanter, three-phase centrifugation, disk-spin centrifugation, filtration centrifugation, decanter centrifugation, Filtration, vacuum filtration, band filter, pressure filtration, membrane filtration, filtration by use of a screen, screen separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, vortex separator or combinations thereof.

In some embodiments, the wet cake may be exposed to one or more wash cycles or washing systems. For example, the wet cake 74 can be processed further if this wet cake 74 is led to a second washing system. In some embodiments, the wet cake 74 can be led to a second mix 60 'and form the wet cake mix 65'. The wet cake mixture 65 'can be led to a second separation 70' and produce a wash centering 75 'and the wet cake 74'. The washing centering 75 'can be recycled to the liquefaction 10. In some embodiments, the washing centering 75' can be combined with the washing centering 75 and recycled to the liquefaction 10. In some embodiments, the wet cake 74 'can be combined with wet cake 74 for further processing as described in the present description. In some embodiments, separation 70 'can be any separation device capable of separating solids and liquids including, for example, centrifugation in bowl decanter, three-phase centrifugation, disk-spin centrifugation, filtration centrifugation, decanter centrifugation , filtration, vacuum filtration, band filter, pressure filtration, membrane filtration, filtration through the use of a screen, screen separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, vortex separator, or combinations thereof. In some embodiments, the wet cake may be exposed to one, two, three, four, five or more wash cycles or washing systems.

The wet cake 74 may be combined with the syrup and then dried by any suitable known process to form DDGS. The formation of the DDGS from the wet cake 74 exhibits various benefits. Because the undissolved solids do not go to the fermenter, the DDGS do not contain trapped extractant and / or alcoholic product, are not subjected to the conditions of the fermenter and do not make contact with the microorganisms present in the fermenter. These advantages facilitate the processing of DDGS, for example, as animal feed.

In some embodiments, a portion of undissolved solids may be conducted until fermentation 30. In some embodiments, this portion of undissolved solids may have smaller particle sizes (eg, fines). In some embodiments, this portion of undissolved solids can form coarse vinasse. In some embodiments, this coarse vinasse can be processed to form light vinasse and a wet cake. In some modalities, the wet cake formed from coarse vinasse and wet cake 74 and / or 74 'may be combined and further processed to produce DDGS.

As shown in Figure 4, the oil is not discharged separately from the wet cake, but the oil is included as part of the wet cake and, ultimately, is present in the DDGS. If corn is used as a raw material, corn oil contains triglycerides, diglycerides, monoglycerides, fatty acids and phospholipids, which provide a source of metabolizable energy for animals. The presence of oil (for example, corn oil) in the wet cake and, finally, in DDGS can provide a desirable food for animals, for example, a high-fat animal feed.

In some embodiments, the oil can be separated from the wet cake and the DDGS, and converted into an ISPR extractant for later use in the same process or other alcoholic fermentation processes. Methods for deriving extractants from biomass are described in the US patent application publication. UU no. 2011/0312044; US patent application publication UU no. 2011/0312043; and publication of US patent application. UU no. 2012/0156738; the complete content of which is incorporated in the present description as a reference. The oil can be separated from the wet cake and the DDGS by the use of any suitable process, which includes, for example, a solvent extraction process. In one embodiment of the invention, the wet cake or DDGS can be added to an extraction unit and washed with a solvent, such as hexane, to remove the oil. Other solvents that can be used include, for example, butanol, isohexane, ethanol, petroleum distillates, such as petroleum ether, or mixtures thereof. After oil extraction, the wet cake or DDGS can be treated to remove residual solvent. For example, the wet cake or DDGS can be heated to evaporate the residual solvent by the use of any method known in the art. After the removal of the solvent, the wet cake or DDGS can be subjected to a drying process to remove residual water. The wet cake processed can be used to generate the DDGS. Processed DDGS can be used as a food supplement for animals, such as dairy animals and cattle, poultry, swine, livestock in general, equine livestock, aquaculture, and domestic pets.

In some embodiments, extractant may be used as a means to modify the color of the wet cake. For example, raw materials such as corn contain pigments (e.g., xanthophylls) that can be used as a coloring agent in food products, including animal feeds (e.g., poultry feeds). The Exposure to extractants can modify these pigments, which results in a lighter colored wet cake, for example. A lighter color of the wet cake may produce DDGS with a lighter color, which may be a desirable quality in certain animal feeds.

In some embodiments where corn is used as a raw material, xanthophylls can be isolated from corn and / or undissolved solids and used as a pigment ingredient in DDGS or animal feed, or as a supplement in pharmaceutical and nutraceutical applications. Methods for isolating xanthophylls include, but are not limited to, chromatography, such as size exclusion chromatography, solvent extraction, such as extraction with ethanol, and enzymatic treatment, such as hydrolysis with alcalase (see, for example, Tsui, et al., J. Food Eng. 83: 590-595, 2007; Li, et al., Food Science 31: 72-77, 2010: U.S. Patent No. 5,648,564, U.S. Pat. No. 6,169,217, U.S. Patent No. 6,329,557, U.S. Patent No. 8,236,929, the entire contents of which are incorporated herein by reference, In some embodiments, xanthophylls can be isolated from the corn and / or undissolved solids and added to COFAs In some embodiments, COFAs and / or xanthophylls can be used for food, pharmaceutical and nutraceutical applications.

After the extraction of the wet cake or DDGS, the Mix obtained from oil and solvent can be collected to separate the oil and the solvent. In one embodiment, the oil / solvent mixture can be processed by evaporation, whereby the solvent is evaporated and can be collected and recielated. The recovered oil can be converted into an ISPR extractant for later use in the same process or another alcoholic fermentation process.

The removal of the oil component from the raw material is advantageous for the production of the alcoholic product, because the oil present in the fermenter can be decomposed into fatty acids and glycerin. Glycerin can accumulate in water and re the amount of water available for recycling throughout the system. Therefore, the removal of the oily component from the raw material can increase the efficiency in the production of alcoholic product by increasing the amount of water that can be recycled through the system.

With reference to Figure 5, the oil can be removed at various points during the processes described in the present description. The suspension of raw material 16 can be separated, for example, with the use of a three-phase centrifuge, in a first liquid phase or aqueous solution 22, a second liquid phase comprising oil 26 and a solid phase or wet cake 24. Wet cake 24 can be further processed to recover fermentable sugars and oil. The wet cake 24 can be led to mixing 60 and combined with water or other solvents to form the wet cake mixture 65. In some embodiments, the water may be countercurrent water, cooking water, process water, Lutter water, evaporation water or any available water source in the fermentation processing facility, or any combination thereof. The wet cake mixture 65 can be conducted to the separation 70 (eg, three phase centrifuge) and produce the washing centering 75, which comprises fermentable sugars, the oil stream 76 and the wet cake 74. The washing centering 75 can recielarse to liquefying 10.

As described in the present description, the wet cake may be exposed to one or more washing cycles or washing systems. In some modalities, the wet cake 74 can be led to a second mix 60 'and form the wet cake mix 65'. The wet cake mixture 65 'can be led to a second separation 70' and produce the wash centering 75 ', the oil stream 76' and the wet cake 74 '. The washing centering 75 'can be recycled to the liquefaction 10. In some embodiments, the washing centering 75' can be combined with the washing centering 75 and recycled to the liquefaction 10. In some embodiments, the wet cake 74 'can be combined with the wet cake 74 for further processing, as described below. In some embodiments, the oil stream 76 'and the oil 26 they can be combined and subjected to further processing for the generation of extractant that can be used in the fermentation process, or the oil stream 76 'and the oil 26 can be combined and subjected to further processing for the manufacture of consumer products.

The wet cake 74 can be combined with the syrup and then dried to form DDGS by the use of any suitable method. The formation of DDGS from wet cake 74 has several advantages. Due to the fact that the solids not dissolved do not go to the fermenter, the DDGS do not contain extractant and / or alcoholic product, are not subjected to the conditions of the fermenter and do not make contact with the microorganisms present in the fermenter. These advantages facilitate the processing of DDGS, for example, as animal feed. As described in the present description, in some embodiments, the wet cake 74, 74 'and the wet cake formed from coarse vinasse can be combined and subjected to further processing to produce DDGS.

As illustrated in Figure 6A, the aqueous solution 22 and the wet cake 24 can be combined, cooled and conducted to the fermentation 30. The suspension of raw material 16 can be separated, for example, with the use of a three-stage centrifuge, in a first liquid phase or aqueous solution 22, a second liquid phase comprising oil 26 and a phase solid or wet cake 24. In some embodiments, the oil 26 may be conducted to a storage tank or any suitable unit for oil storage. The aqueous solution 22 and the wet cake 24 can be conducted to the mixture 80 and resuspended to form the aqueous solution / wet cake mixture 82. The mixture 82 can be conducted to the cooler 90 and produce the cooled mixture 92 that can be conducted to the fermentation. In some embodiments, when the oil 26 is removed from the raw material suspension 16 by means of the separation 20, the mixtures 82 and 92 include a reduced amount of oil.

In another embodiment, as illustrated in Figure 6B, the suspension of raw material 16 can be separated by the use of a separation device (eg, a three-phase centrifuge) to generate a first liquid phase or aqueous solution 22, a second liquid phase comprising the oil 26 and a solid phase or wet cake 24. The aqueous solution 22, the wet cake 24 and the oil 26, or portions thereof, can be conducted to the fermentation 30. In some embodiments, the aqueous solution 22, wet cake 24 and oil 26, or portions thereof, may be combined, for example, by mixing, to form a mixture of aqueous solution, wet cake and oil, and the mixture may be conducted to fermentation 30. In some embodiments, the aqueous solution 22 and the wet cake 24 can be combined to form a mixture of aqueous solution and wet cake; then, the oil 26 can be added to the mixture to form a mixture of aqueous solution, wet cake and oil, and this mixture can be conducted to the fermentation 30. In some embodiments, the aqueous solution 22 and the wet cake 24 can be combined to form a mixture of aqueous solution and wet cake, and this mixture and the oil 26, or a portion thereof, can be conducted to the fermentation 30 as separate streams.

In additional embodiments of the processes and systems described in the present disclosure, saccharification can take place in a separate saccharification system. In some embodiments, a saccharification system may be located between liquefaction 10 and separation 20, or between separation 20 and fermentation 30. In some embodiments, liquefaction and / or saccharification may be performed with the use of crude starch enzymes or low temperature hydrolysis enzymes such as Stargen ™ (Genencor International, Palo Alto, CA) and BPX ™ (Novozymes, Franklinton, NC). In some embodiments, the suspension of raw material may be exposed to hydrolysis of crude starch (also known as cold cooking or cold hydrolysis).

In some embodiments, the systems and processes of the present invention may include a series of two or more separation devices (eg, centrifuges) for the removal of undissolved solids and / or oil. For example, a aqueous solution discharged from a first separation unit can be conducted to an inlet of a second separation unit. The first separation unit and second separation unit can be identical (for example, two three-phase centrifuges) or they can be different (for example, a three-stage centrifuge and a decanter centrifuge). The separation can be achieved by a number of means including, but not limited to, centrifugation decanter bowl centrifugation three phase centrifugation packed, centrifugation filtration, decanter centrifugation, filtration, vacuum filtration, filter band, pressure filtration, membrane filtration, filtration by using a sieve, separation sieve separation grid separation porous grid, flotation, hidrocielón, filter press, screw press, gravity settler, separator Vortex, or combinations of these.

The absence or minimization of undissolved solids in the fermentation broth has several advantages. For example, the need for operating units in downstream processing can be eliminated, which results in an increase in efficiency for the production of the alcoholic product. In addition, some or all of the centrifuges used to process the coarse vinasse can be eliminated, since there are less undissolved solids in the fermentation broth that comes out of the fermenter. The removal of undissolved solids from the raw material suspension can improve the productivity of biomass processing and profitability. Better productivity can include better efficiency in the production of the alcoholic product and / or an increase in extraction activity with respect to processes and systems that do not remove the dissolved solids before fermentation. See another description of the processes and systems for separating undissolved solids from the suspension of raw material in, for example, the publication of US patent application. UU no. 2012/0164302 and the international patent application of PCT no. PCT / US2013 / 51571, the complete contents of which are incorporated herein by reference.

As described in the present description, the alcoholic product can be recovered from the fermentation broth with the use of a number of methods, including liquid-liquid extraction. In some embodiments of the processes and systems described in the present disclosure, an extractant may be used to recover the alcohol product from the fermentation broth. The extractants used herein may, for example, one or more of the properties and / or following characteristics: (i) biocompatible with microorganisms, (ii) immiscible with the fermentation broth, (iii) a high coefficient of partition (K p) for the extraction of the alcoholic product, (iv) a low partition coefficient for the extraction of nutrients and / or water, (v) low viscosity (m), (vi) high selectivity for the alcoholic product in comparison with, for example, water, (vii) low density (p) with respect to the fermentation broth or a different density compared to the density of the fermentation broth, (viii) a suitable boiling point for the downstream processing of the extractant and the alcoholic product, (ix) a melting point lower than the ambient temperature, (x) minimum absorbance in solids, (xi) a low tendency to form emulsions with the fermentation broth, (xii) stability throughout the fermentation process, (xii) ) low cost and (xiv) not dangerous.

In some embodiments, the extractant may be selected on the basis of certain properties and / or characteristics, as described in the present description. For example, the viscosity of the extractant can influence the mass transfer properties of the system, i.e., the efficiency with which the alcohol product can be extracted from the aqueous phase to the extractant phase (i.e., the organic phase). The density of the extractant can affect the phase separation. In some embodiments, selectivity refers to the relative amounts of alcoholic product relative to the water absorbed by the extractant. The boiling point can affect the cost and method of recovery of the alcoholic product. For example, in the case where butanol is recovered from the extractant phase by distillation, the boiling point of the extractant should be low enough to allow separation of butanol while minimizing thermal degradation or secondary reactions of the extractant, or the need for high vacuum in the distillation process.

The extractant may be biocompatible with the microorganism, i.e., non-toxic to the microorganism or toxic only to the extent that the microorganism deteriorates to an acceptable level. In some embodiments, biocompatible refers to the measurement of a microorganism's ability to use fermentable carbon sources in the presence of an extractant. The degree of biocompatibility of an extractant can be determined, for example, by the rate of use of the glucose of the microorganism in the presence of the extractant and the alcoholic product. In some embodiments, a non-biocompatible extractant refers to an extractant that interferes with the ability of a microorganism to use fermentable carbon sources. For example, a non-biocompatible extractant does not allow the microorganism to use glucose at a rate greater than about 25%, greater than about 30%, greater than about 35%, greater than about 40%, greater than about 45% or greater than about 50% of the speed when the extractant is not present.

A person skilled in the art can select an extractant to maximize the desired properties and / or characteristics as described in the present description and to optimize the recovery of an alcoholic product. In addition, a person skilled in the art can appreciate that the use of a mixture of extractants can be advantageous. For example, mixtures of extractants can be used to increase the partition coefficient of the alcoholic product. In addition, mixtures of extractants can be used to adjust and optimize the physical characteristics of the extractant, such as density, boiling point and viscosity. For example, the appropriate combination can provide an extractant having a sufficient alcohol product partition coefficient and sufficient biocompatibility to allow its economical use to remove the alcohol product from the fermentation broth.

In some embodiments, extractants useful in the processes and systems described in the present disclosure may be organic solvents. In some embodiments, the extractants useful in the processes and systems described in the present disclosure may be organic solvents immiscible in water. In some embodiments, the extractant may be an organic extractant selected from the group consisting of saturated, monounsaturated fatty alcohols and C12 to C22 polyunsaturates, C12 to C22 fatty acids, C12 to C22 fatty acid esters, C12 to C22 fatty aldehydes, C12 to C22 fatty amides, and mixtures thereof. In some embodiments, the extractant may also be an organic extractant selected from the group consisting of saturated, monounsaturated and polyunsaturated fatty alcohols of C4 to C22, C4 to C2a fatty acids, fatty acid esters of C4 to C28, fatty aldehydes. from C4 to C22, fatty amides of C4 to C22 and mixtures of these. In some embodiments, the fatty acid may a C4 to C24 fatty acid and / or the aster may be an ester of a C4 to C24 fatty acid. In some embodiments, the extractant may be an organic extractant selected from the group consisting of saturated, monounsaturated and polyunsaturated fatty alcohols of C12 to Cie, C12 to Cie fatty acids, fatty acid esters of C12 to Cis, C12 fatty aldehydes to Cie, fatty amides of C12 to Cie, and mixtures of these. In some embodiments, the extractant may be an organic extractant selected from the group consisting of saturated, monounsaturated and polyunsaturated fatty alcohols of Ci4 to Cie, Ci4 to Cie fatty acids, Ci4 to Cie fatty acid esters, Ci4 fatty aldehydes to Cie, Ci to Cie fatty amides, and mixtures of these. In some embodiments, the extractant may be an organic extractant selected from the group consisting of saturated, monounsaturated and polyunsaturated fatty alcohols of Ci6 to Cis, fatty acids from Cie to Cis, fatty acid esters of Ci6 to Cis, fatty aldehydes from Cie to Ci8, fatty amides of Ci6 to Cie, and mixtures of these. In some embodiments, the extractant may comprise carboxylic acids. In some embodiments, the aster of a fatty acid may be the combination of a fatty acid with an alcohol (e.g., fatty ester). In some modalities, alcohol can be an alcoholic product. In some embodiments, the aster may be methyl ester, ethyl ester, propyl ester, butyl ester, pentyl ester, hexyl ester, or glyceride.

In some embodiments, the extractant may include a first extractant selected from Ci2 to C12 fatty alcohols, C12 to C12 fatty acids, C12 to C12 fatty acid esters, C12 to C12 fatty aldehydes, C12 to C12 fatty amides, and mixtures of these; and a second extractant selected from C12 to C22 fatty alcohols, C12 to C22 fatty acids, C12 to C22 fatty acid esters, C12 to C22 fatty aldehydes, C12 to C22 fatty amides, and mixtures thereof. In some embodiments, the extractant may include a first extractant selected from C 12 to C 22 fatty alcohols, C 12 to C 22 fatty acids, C 12 to C 22 fatty acid esters and mixtures thereof; and a second extractant selected from C12 to C22 fatty alcohols, C12 to C22 fatty acids, C12 to C22 fatty acid esters and mixtures thereof. In some embodiments, the extractant may include a first extractant selected from Ci2 to Ci8 fatty alcohols, C12 to Ci8 fatty acids / Ci2 to Ci8 fatty acid esters and mixtures thereof; and a second extractant selected from Ci2 to Ci8 fatty alcohols, Ci2 to Ci8 fatty acids, Ci2 to Ci8 fatty acid esters, and mixtures thereof. In some embodiments, the extractant may include a first extractant selected from Ci4 to Ci8 fatty alcohols, Ci4 to Ci8 fatty acids, Ci4 to Ci8 fatty acid esters, and mixtures thereof; and a second extractant selected from fatty alcohols of Ci4 to Ci8i, Ci4 to Ci8 fatty acids, Ci4 to Ci8 fatty acid esters, and mixtures thereof. In some embodiments, the extractant may include a first extractant selected from Ci2 to Ci2 fatty alcohols, Ci2 to Ci2 fatty acids, Ci2 to Ci2 fatty acid esters, Ci2 to Ci2 fatty aldehydes, Ci2 to Ci2 fatty amides, and mixtures of these; and a second extractant selected from fatty alcohols of C7 to Cu, C7 to Cu fatty acids, fatty acid esters of C7 to Cu, C7 to Cu fatty aldehydes, and mixtures of these.

In some embodiments, the extractant may be an organic extractant, such as oleyl alcohol, behenyl alcohol, cetyl alcohol, lauryl alcohol (also known as 1-dodecanol), myristyl alcohol, stearyl alcohol, oleic acid, lauric acid, linoleic acid. , linolenic acid, myristic acid, palmitic acid, acid stearic acid, octanoic acid, decanoic acid, undecanoic acid, methyl myristate, methyl oleate, 1-nonanol, 1-decanol, 2-undecanol, 1-nonanal, 1-undecanol, undecanal, lauric aldehyde, 2-methylundecanal, oleamide, linoleamide, palmitamide, stearylamide, 2-ethyl-1-hexanol, 2-hexyl-1-decanol, 2-octyl-1-dodecanol, and mixtures thereof. In some embodiments, the extractant may comprise one or more of the following: oleic acid, lauric acid, linoleic acid, linolenic acid, myristic acid, palmitic acid, stearic acid, octanoic acid, decanoic acid, and undecanoic acid. In some embodiments, the extractant may comprise one or more of the following: oleic acid, linoleic acid, linolenic acid, myristic acid, palmitic acid, and stearic acid. In some embodiments, the extractant may comprise one or more of the following: oleic acid, linoleic acid, palmitic acid, and stearic acid. In some embodiments, the extractant may comprise one or more of the following: oleic acid, lauric acid, linoleic acid, linolenic acid, myristic acid, palmitic acid, stearic acid, octanoic acid, decanoic acid and undecanoic acid, and one or more esters of oleic acid, lauric acid, linoleic acid, linolenic acid, myristic acid, palmitic acid, stearic acid, octanoic acid, decanoic acid, and undecanoic acid. In some embodiments, the extractant may comprise one or more of the following: oleic acid, linoleic acid, linolenic acid, myristic acid, palmitic acid and stearic acid, and one or more esters of oleic acid, linoleic acid, linolenic acid, myristic acid, palmitic acid, and stearic acid. In some embodiments, the extractant may comprise one or more of the following: oleic acid, linoleic acid, palmitic acid and stearic acid, and one or more esters of oleic acid, linoleic acid, palmitic acid, and stearic acid. In some embodiments, the extractant may comprise one or more of the following: oleyl alcohol, behenyl alcohol, cetyl alcohol, lauryl alcohol, myristyl alcohol, stearyl alcohol. In some embodiments, the extractant may comprise one or more of the following: 1-nonanol, 1-decanol, 2-undecanol, 1-nonanal, 1-undecanol, undecanal, 2-ethyl-1-hexanol, 2-hexyl-1 - decanol, 2-octyl-l-dodecanol.

In some embodiments, the extractant may be a mixture of biocompatible and non-biocompatible extractants. Examples of mixtures of biocompatible and non-biocompatible extractants include, but are not limited to, oleyl alcohol and nonanol, oleyl alcohol and 1-undecanol, oleyl alcohol and 2-undecanol, oleyl alcohol and 1-nonanal, oleyl alcohol and decanol, and oleyl alcohol and dodecanol. Other examples of biocompatible and non-biocompatible extractants are described in the US patent application publication. UU no. 2009/0305370 and the publication of the patent application of the USA UU no. 2011/0097773; whose complete contents are incorporated in the present description as a reference. In some embodiments, biocompatible extractants may have high atmospheric boiling points. For example, biocompatible extractants can have boiling points at atmospheric pressure greater than the atmospheric boiling point of water.

In some embodiments, a hydrophilic solute may be added to the fermentation broth which is contacted with an extractant. The presence of a hydrophilic solute in the aqueous phase can improve the phase separation and can increase the fraction of alcoholic product that is partitioned to the organic phase. Examples of hydrophilic solute may include, but are not limited to, polyhydroxy compounds, polycarboxylic compounds, polyol compounds and dissociated ionic salts. Sugars such as glucose, fructose, sucrose, maltose and oligosaccharides can serve as a hydrophilic solute. Other polyhydroxy compounds may include glycerol, ethylene glycol, propanediol, polyglycerol, and hydroxylated fullerene. The polycarboxylic compounds may include citric acid, tartaric acid, maleic acid, succinic acid, polyacrylic acid, and sodium, potassium, or ammonium salts thereof. The ionic salts that can be used as a hydrophilic solute in the fermentation broth include cations including sodium, potassium, ammonium, magnesium, calcium and zinc; and anions which include sulfate, phosphate, chloride and nitrate. A person skilled in the art can select the amount of hydrophilic solute in the fermentation broth to maximize the transfer of alcoholic product from the aqueous phase (eg, fermentation broth) to the organic phase (eg, extractant) without having a negative impact on the growth and / or productivity of the microorganism that produces the alcoholic product. High levels of hydrophilic solute may impose osmotic stress and / or toxicity on the microorganism. A person skilled in the art can use any number of known methods to determine an optimum amount of hydrophilic solute to minimize the effects of osmotic stress and / or toxicity in the microorganism.

In some embodiments where the alcoholic product is butanol, the extractant may be selected to attract the alkyl portion of the butanol and to provide little or no affinity for the water. An extractant that does not offer hydrogen bonds, for example, to water, will absorb alcohol selectively. In some embodiments, the extractant may comprise an aromatic compound. In some embodiments, the extractant may comprise alkyl substituted benzenes including, but not limited to, eumeno, para-cymene (also known as 1-methyl-4- (1-methylethyl) benzene), meta-cymene ( known, in addition, such as 1- methyl-3- (1-methylethyl) benzene), meta-diisopropylbenzene, para-diisopropylbenzene, triethylbenzene, ethyl butyl benzene and tert-butylstyrene. An advantage of using a benzene substituted with alkyl is that the affinity for butanol is comparatively greater with respect to other hydrocarbons. In addition, benzenes substituted with isopropyl or isobutyl may offer a particular advantage of the affinity for butanol with respect to other substituted benzenes. Another advantage is the lower viscosity, lower surface tension, lower density, greater thermal stability and greater chemical stability, which help phase separation and long-term reuse. In some embodiments, an extractant that attracts the alkyl portion of butanol may be combined with another extractant that offers affinity in the form of hydrogen bonds, for example, with the hydroxylic portion of butanol, so that the mixture provides an optimum balance between selectivity and partition with respect to water. In some embodiments, an extractant containing butanol can be separated from the fermentation broth by phase separation and distilled in a column that functions under vacuum. This distillation can work with reflux to maintain a high purity butanol distillate containing very little extractant. The lower part may comprise a portion of the butanol contained in the distillation feed, so that the Reboiling temperature is suitable for the indirect supply of heat from the available steam. The distillation can be carried out with a partial coolant, where only the reflux liquid is condensed, and a steam distillate, the composition of which is substantially butanol, can be directed to the lower part of a rectification column which is simultaneously fed with a butanol stream decanted from the condensed vapor of the beer at the top of the column. An advantage of this type of distillation is that the heat that integrates the steam generated by desorbing the butanol from the extractant eliminates the need for a steam boiler to purify the decanted butanol stream.

In some embodiments, extractant can be generated from raw material. For example, oils, such as corn oil, present in the raw material can be used to generate extractant for extractive fermentation. The glycerides in the oil can be converted chemically or enzymatically into a reaction product, such as fatty acids and / or fatty esters (for example, ethyl esters, butyl esters, fusel esters) that can be used as an extractant for recovery of the alcoholic product. With the use of corn oil as an example, triglycerides from corn oil can react with a base, such as ammonium hydroxide, to obtain amides fats and glycerol. In some embodiments, the oil from the raw material can be hydrolyzed by a catalyst to generate fatty acids. In some embodiments, at least a portion of the acylglycerides of the oil can be hydrolyzed to carboxylic acid by contacting the oil with a catalyst. In some embodiments, the resulting acid / oil composition includes monoglycerides and / or diglycerides from the partial hydrolysis of the acylglycerides in the oil. In some embodiments, the resulting acid / oil composition includes glycerol, a by-product of the hydrolysis of the acylglyceride. In some embodiments, the acid / oil composition obtained includes lysophospholipids derived from the partial hydrolysis of the phospholipids of the oil. Methods for deriving extractants from biomass are described in the US patent application publication. UU no. 2011/0312044; US patent application publication UU no. 2011/0312043; and the publication of US patent application. UU no. 2012/0156738, whose complete contents are incorporated in the present description as a reference.

In some embodiments, the conversion of the oil (eg, hydrolysis, transesterification) of the raw material or suspension of raw material can take place in the thermistor by the addition of a catalyst to the triturator. For example, a catalyst, such as as lipase, to the thermistor and convert the oil present in the raw material or raw material in suspension in fatty acids and / or fatty esters. In some embodiments, the conversion of the raw material oil or suspension of raw material can take place in a separate unit. For example, the raw material or suspension of raw material can be conducted in a unit, a catalyst, such as lipase, can be added to the unit and the oil present in the raw material or raw material suspended in fatty acids can be converted. As another example, the raw material or suspension of raw material can be conducted in one unit, a catalyst, such as lipase, and an alcohol (for example, ethanol, butanol, fusel alcohols) can be added to the unit and the oil present can be converted in the raw material or suspension of raw material in fatty esters. In some embodiments, the fatty acids and / or fatty esters can be added to the fermenter and can be used as an extractant for the recovery of the alcoholic product. In some embodiments, the fatty acids and / or fatty esters can be added to an external extractor or extractant column and can be used as an extractant for the recovery of the alcoholic product.

In some embodiments, the oil can be separated from the raw material suspension and can be conducted to a unit, and a catalyst, such as lipase, can be added to the unit to generate a fatty acid stream. The current of The fatty acid can be heated to deactivate the lipase, and then the fatty acid stream can be conducted to an external extractor or a storage tank. The fatty acids from the storage tank can be conducted to an external extractor to extract the alcoholic product from the fermentation broth. In some embodiments, the oil separated from the raw material suspension can be stored in a storage tank. A catalyst, such as lipase, can be added to the storage tank to generate a fatty acid stream. The fatty acid stream can be heated to deactivate the lipase, cooled, and then conducted to an external extractor to extract the alcoholic product from the fermentation broth. In some embodiments, the oil separated from the raw material suspension can be conducted to a unit, and a catalyst, such as lipase, can be added to the unit to generate a fatty acid stream. The fatty acid stream can be heated to deactivate the lipase, cooled, and then the fatty acid stream can be conducted to a scrubber.

In some embodiments, the one or more catalysts may be one or more enzymes, for example, hydrolase enzymes. In some embodiments, the one or more catalysts may be one or more enzymes, for example, lipase enzymes. Lipase enzymes can be derived from any source, which includes, for example, Absidia, Achromobacter, Aeromonas, Alcaligenes, Alternaria, Aspergillus, Achromobacter, Aureobasidium, Bacillus, Beauveria, Brochothrix, Candida, Chromobacter, Coprinus, Fusarium, Geotricum, Hansenula, Humicola, Hyphozyma, Lactobacillus, Metarhizium, Mucor, Nectria, Neurospora, Paecilomyces, Penicillium, Pseudomonas, Rhizoctonia, Rhizomucor, Rhizopus, Rhodosporidium, Rhodotorula, Saccharomyces, Sus, Sporobolomyces, Thermomyces, Thiarosporella, Trichoderma, Verticillium and / or Yarrowia. In some embodiments, the lipase source can be selected from the group consisting of Absidia blakesleena, Absidia corymbifera, Achromobacter iophagus, Alcaligenes sp., Alternaria brassicola, Aspergillus flavus, Aspergillus niger, Aspergillus tubingensis, Aureobasidium, Bacillus coagulans pullulans, Bacillus pumilus, Bacillus strearothermophilus, Bacillus subtilis, Brochothrix thermosohata, Burkholderia cepacia, Candida cylindracea (Candida rugosa), Candida paralipolytica, Candida antarctic lipase A, Candida antarctic lipase B, Candida ernobii, Candida deformans, Candida rugosa, Candida parapsilosis, Chromobacter viscosum, Coprinus cinerius, Fusarium heterosporum, Fusarium oxysporum, Fusarium solani, Fusarium solani pisi, Fusarium roseum culmorum, Geotrichum candidum, Geotricum penicillatum, Hansenula an mala, Humicola brevispora, Humicola brevis var. thermoidea, Humicola insolens, Lactobacillus curvatus, Rhizopus oryzae, Mucor javanicus, Neurospora crassa, Nectria haematococca, Penicillium cyclopium, Penicillium crustosum, Penicillium expansum, Penicillium roque f orti, Penicillium camembertii, Penicillium sp. I, Penicillium sp. II, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas cepacia (syn. Burkholderia cepacia), Pseudomonas fluorescens, Pseudomonas fragi, Pseudomonas maltophilia, Pseudomonas mendocina, Pseudomonas mephitica lipolytica, Pseudomonas alcaligenes, Pseudomonas plantari, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri and Pseudomonas wisconsinensis, Rhizoctonia solani, Rhizomucor miehei, Rhizopus arrhizus, Rhizopus delemar, Rhizopus j japonicus, Rhizopus microsporus, Rhizopus nodosus, Rhizopus oryzae, Rhodosporidium toruloides, Rhodotorula glutinis, Saccharomyces cerevisiae, Sporobolomyces shibatanus, Sus scrofa, Thermomyces lanuginosus (exHumicola lanuginose), Thiarosporella phaseolina, Trichoderma harzianum, Trichoderma reese i and Yarrowia lipolytica.

In some embodiments, the hydrolase and / or lipase can be expressed by the microorganism. In some embodiments, the microorganism can be genetically engineered to express the homologous or heterologous hydrolase and / or lipase. In some embodiments, hydrolase and / or lipase can be expressed by a microorganism that produces, in addition, an alcoholic product. In some embodiments, the hydrolase and / or lipase can be expressed by a microorganism that also expresses a butanol biosynthetic pathway. In some embodiments, the butanol biosynthetic pathway may be a biosynthetic pathway of 1-butanol, 2-butanol biosynthetic pathway, isobutanol biosynthetic pathway, or 2-butanone biosynthetic pathway.

Commercial preparations of lipase suitable as a catalyst include, but are not limited to, Lipolase® 100 L, Lipex® 100L, Lipoclean® 2000T, Lipozyme® CALB L, Novozyme® CALA L and Palatase 20000L, available from Novozymes (Franklinton, NC) , or lipases from Pseudomonas fluorescens, Pseudomonas cepacia, Mucor miehei, pig pancreas, Candida cylindracea, Candida rugosa, Rhizopus niveus, Candida anticactica, Rhizopus arrhizus or Aspergillus, available from Sigma Aldrich (St. Louis, MO). In some embodiments, the lipase may be thermostable and / or heat tolerant, and / or solvent tolerant.

In some embodiments, the one or more catalysts may be phospholipases. A phospholipase useful in the present invention can be obtained from a variety of biological sources, for example, but not limited to, filamentous fungal species within the genus Fusarium, such as a strain of Fusarium culmorum, Fusarium heterosporum, Fusarium solani or Fusarium oxysporum; or a filamentous fungal species within the genus Aspergillus, such as a strain of Aspergillus awamori, Aspergillus foetidus, Aspergillus japonicus, Aspergillus niger or Aspergillus oryzae. Also useful in the present invention are the phospholipase variants of Thermomyces lanuginosus, such as the commercial product Lecitase® Ultra (Novozymes A'S, Denmark). One or more phospholipases can be applied as lyophilized powders, immobilized or in aqueous solution.

In some embodiments, the phospholipase can be expressed by the microorganism. In some embodiments, the microorganism can be genetically engineered to express homologous or heterologous phospholipases. In some embodiments, the phospholipase can be expressed by a microorganism that also produces an alcoholic product. In some embodiments, the phospholipase can be expressed by a microorganism that also expresses a butanol biosynthetic pathway. In some embodiments, the butanol biosynthetic pathway may be a biosynthetic pathway of 1-butanol, 2-butanol biosynthetic pathway, isobutanol biosynthetic pathway, or 2-butanone biosynthetic pathway.

Fermentation by-products such as isobutyric acid, phenylethanol, 3-methyl-1-butanol, 2-methyl-1-butanol, isobutyraldehyde, acetic acid, ketoisovaleric acid, pyruvic acid and dihydroxyisovaleric acid may have an inhibitory effect on the microorganism. In some embodiments, these by-products can be modified by esterification. By For example, the by-products can be esterified with carboxylic acids, alcohols, fatty acids or other by-products. In some embodiments, these esterification reactions may be catalyzed by lipases or phospholipases. As an example, the lipase present in the fermentation broth can catalyze the esterification of the by-products generated during the fermentation. The esterification of these by-products can minimize their inhibitory effects on the microorganism.

With reference to Figure 7A, raw material 12 can be processed as described in Figures 1 to 6 and, therefore, will not be described in detail. The aqueous solution 22 can be subjected to further treatment to remove the residual oil. In some embodiments, the aqueous solution 22 may be subjected to centrifugation, decanting or any other method that may be used for the removal of the oil. In some embodiments, the aqueous solution 22 can be conducted to the unit 25 (or vessel) and the catalyst 23 (eg, lipase) can be added to the unit 25 to convert the oil present in the aqueous solution 22 into fatty acids and generate the stream 27. Then, stream 27 can be conducted to fermentation 30 and microorganism 32 can be added, in addition, to fermentation 30 to produce the alcoholic product. After fermentation 30, the stream 31 comprising the alcoholic product and fatty acids can be conducted to an external unit, for example, a external extractor or external extraction circuit for the recovery of the alcoholic product.

With reference to Figure 7B, in some embodiments, the catalyst 23 can be deactivated, for example, by heating. In some embodiments, the stream 27 comprising the catalyst 23 can be heated (q) to deactivate the catalyst 23 before the addition to the fermentation 30. With reference to Figure 7C, in some embodiments, deactivation can be performed in a separate unit , for example, a deactivation unit. In some embodiments, stream 27 can be conducted to deactivation 28. After deactivation, stream 27 'can be conducted to fermentation 30 and microorganism 32 can be added, in addition, to fermentation 30 to produce the alcoholic product.

The extraction of the oil from the aqueous solution 22 through the conversion of the oil into fatty acids can translate into energy savings for the production plant due to a more efficient fermentation, less obstruction of the equipment due to oil removal, lower energy requirements , for example, the energy needed to dry the distillers grains and a better functioning of the evaporators or the evaporation train. In addition, the removal of the oily component of the raw material is advantageous for the production of the alcoholic product, because the oil present in the thermenator it can be broken down into fatty acids and glycerin. Glycerin can accumulate in the water and reduce the amount of water available for recielado through the system. Therefore, the removal of the oil component from the raw material increases the efficiency of the production of the alcohol product by increasing the amount of water that can be recycled through the system. In addition, the removal of oil decreases the likelihood of stable emulsions. In some embodiments of the present invention, in the event that an emulsion is formed, it can be easily broken by mechanical processing, addition of protic solvents or other conventional means.

In another embodiment, with reference to Figure 7D, aqueous solution 22 can be conducted to fermentation 30 and catalyst 23 (eg, lipase) can be added to fermentation 30 to convert the oil present in aqueous solution 22 into fatty acids. and / or fatty esters. In some embodiments, the fatty esters can be derived from the combination of fatty acids with an alcohol. In some embodiments, the alcohol can be any alcohol of fermentation 30, which includes an alcoholic product. In some embodiments, the amount of oil in the aqueous solution 22 converted into fatty acids and / or fatty esters can be at least about 50%, at least about 60%, at least about 70%, at least about 80%, at less about 90% or at least about 95%. In some embodiments, the proportion of fatty esters and fatty acids generated by the conversion of the oil may be approximately 75:25. In some embodiments, the ratio of fatty esters and fatty acids can be approximately 80:20. In some embodiments, the catalyst 23 can be added to the fermentation 30 in an amount such as to maintain a certain rate of oil conversion.

After fermentation 30, the stream 31 comprising alcoholic product, fatty acids and fatty esters may be subjected to further processing for the recovery of the alcoholic product. For example, the stream 31 may be conducted to an external unit, for example, an external extractor or external extraction circuit for the recovery of the alcoholic product. In some embodiments, the fatty acids and fatty esters of stream 31 can be used as an extractant. In some embodiments, the external unit may comprise extractant. In some embodiments, the extractant may comprise fatty acids and / or fatty acid esters.

The present invention also provides processes and systems for recovering an alcohol product produced by a fermentative process. One of those processes to recover an alcoholic product is liquid-liquid extraction. The use of liquid-liquid extraction as an ISPR technique works best with an extraction process liquid-liquid that maximizes the net present value of the capital investment necessary for the practice of technology. One aspect of maximizing the net present value of a liquid-liquid extraction process is to avoid the large operating and capital cost expenses associated with separating the extractant from the fermentation broth.

In one embodiment of a liquid-liquid extraction process, the extractant may be added directly to the fermenter, and the fermentation broth and the extractant may be mixed together in a manner that affects mass transfer (eg, transfer of the alcohol product from the fermenter). fermentation broth to the extractant) and allow the fermentation to be produced at a highly effective alcoholic product title. In that process, if the mixing is too intense or vigorous, it is possible that the fermentation broth and the extractant should be separated by the use of a separation device, such as a centrifuge. If the mixing is not too intense, phase separation can be achieved by gravity sedimentation caused by the density difference between the extractant and the fermentation broth. In any case, additional thermomentors may be required to overcome the volume loss of the thermistor occupied by the extractant added to the thermistor. The addition of extractant directly to the burner can be done in the modes batch, semilote or continuous, regardless of the separation of phases inside the thermoretor. If the continuous mode is used and separation by gravity of the fermentation broth and the extractant is not possible, then a separation device such as a centrifuge may be required to separate the alcohol product from the extractant. If the separation process used to remove the alcoholic product from the extractant is such that the microorganism present in the fermentation broth is viable the separation process, then the separation of the fermentation broth from the alcoholic / extractant product may not be necessary.

Another embodiment of a liquid-liquid extraction process may include an external extractor or an extraction column. For example, the fermentation broth of the fermenter can be conducted to an external extractor, where the fermentation broth is mixed with the extractant. Then, the mixture of fermentation broth and extractant can be separated to generate a stream of fermentation broth more depleted in alcoholic product and an extractant stream more enriched in alcoholic product. The most impoverished fermentation broth stream can return to the fermenter. The more enriched extractant stream can be subjected to further processing to remove at least a portion of the alcohol product from the extractant for product recovery. alcoholic In some embodiments, the rate of recovery of the alcoholic product from the extractant stream may be adjusted at a rate to maintain the production of the plant. In some embodiments, the liquid-liquid extraction process may comprise one or more external liquid-liquid extractors.

In some embodiments, the fermentation may take place in the thermodor and the external extractor. The additional volume of fermentation broth present in the external extractor can serve to increase the total volume of the fermenter and, therefore, can increase the total production of the alcoholic product.

The performance of the external extractor with respect to the removal of the alcoholic product may depend on the surface available for interfacial contact, the physical nature of the fermentation broth and the extractant, the relative amounts of the two phases (eg, broth phase). fermentation and phase of the extractant) present in the external extractor, and the concentration difference of the driving force between the phases of the fermentation broth and the extractant. The maximization of the effncy of the external extractor for a given driving force of concentration of the alcoholic product can be effected by reducing the droplet size of the dispersed phase in the external extractor, for example, by means of the design of the nozzle, design of the internal components and / or agitation. In some embodiments, the design and operation of the external extractor can provide suffnt mixing to effect proper transfer of the alcoholic product between the phases of the fermentation broth and the extractant, to maintain the productivity requirements of the alcoholic product.

In some cases, the CO2 from the fermentation can be generated in the external extractor, which leads to the formation of droplets that can interfere with the separation of phases. For example, the droplets of the fermentation broth can adhere to the CO2 that is released from the phase of the extractant. In some embodiments, the extractant phase can be maintained as the continuous phase to improve coalescence of the droplets. In some modalities, the external extractor may include internal components or output ports for CO2. For example, a coalescence filter may be added to the external extractor, and / or the output ports may be located to improve coalescence and recovery of the fermentation broth phase.

The conditions for separating the alcoholic product from the fermentation broth can be detrimental to the microorganism present in the fermentation broth. In some embodiments, the microorganism can be separated from the fermentation broth before contacting the fermentation broth with the extractant. In some modalities, the The microorganism can be separated from a mixture of fermentation broth and extractant before separation (or processing) of this mixture. Any separation method capable of separating the microorganism from the fermentation broth or from the fermentation and extractant broth mixture, including, for example, centrifugation, can be used. By separating the microorganism before contacting the fermentation broth with the extractant, it may be possible to use more stringent extraction conditions, such as higher temperatures and / or non-biocompatible extractants. If a separation method not harmful to the microorganism was used, then the separation of the fermentation broth and the extractant before the removal of the alcoholic product may not be required.

If the extractant and the fermentation broth do not separate, then the extractant can be included in the feed of the evaporator train and, therefore, become a component of the syrup formed during evaporation and, possibly, incorporated into animal feed. . In some embodiments, the extractant can be separated from the syrup by the use of any means of separation, including, for example, centrifugation. A low-boiling biocompatible extractant (for example, compared to water) may not require that separation, because the extractant and water may recielarse to use in the production process.

In a typical corn-to-alcoholic production plant, the water balance of the total production process can be maintained by recycling the water from the production plant with recycled water distilled in an evaporator train to remove salts and other dissolved solids from the water. beer. The syrup obtained from the evaporator train can be mixed with undissolved solids, and the mixture can be dried and marketed as animal feed. Processes and systems for processing undissolved solids for animal feed are described, for example, in the US patent application publication. UU no. 2012/0164302; US patent application publication UU no. 2011/0315541; US patent application publication UU no. 2013/0164795; and PCT international patent application no. PCT / US2013 / 51571, the complete contents of which are incorporated herein by reference.

As described in the present description, undissolved solids can be removed from the raw material (or suspension of raw material) before adding the raw material to the fermentation. If the undissolved solids are not removed upstream of the fermentation, then the centrifugation of the beer may be required to remove the undissolved solids, in order to avoid obstruction. of the evaporators. For example, in a commercial corn production plant that is dry and ground to an alcoholic product, the content of undissolved solids in an evaporator train feed can operate at approximately 3% of the total solids in suspension and may have 3.5-4% of total solids in suspension. An upstream process that removes enough solids to keep the percentage of total solids in suspension at or below these percentage values can eliminate the need for centrifugation, for example, before driving the beer to the evaporators (or to the evaporation train). The elimination of this centrifugation would result in a saving in the capital needed to modernize a plant producing corn that is dry and ground to an alcoholic product.

By removing at least a portion of the undissolved solids present in the raw material suspension prior to fermentation, the interfacial surface area between the phases of the fermentation broth and the extractant can be increased in an external extractor by reducing the amount of undissolved solids at the interface, which improves the transfer of the alcoholic product between the fermentation broth and the extractant, and provides a net phase separation between the fermentation broth and the extractant. A clear phase separation can also eliminate the need for additional separation steps (for example, centrifugation) and, therefore, a saving in capital expenditures.

The separation of the fermentation broth and the extractant leaving the external extractor may be influenced by the solids content and the particle size distribution of the solids content in the fermentation broth, the gas content and the bubble size distribution of gas in the fermentation broth, the physical properties of the fermentation broth and the extractant, which include, but they are not limited to, viscosity, density and surface tension, as well as the design and operation of the external extractor, and the design and operation of the thermoretor. These properties may determine the need for separation devices (eg, centrifuges) to separate the fermentation broth and the extractant leaving the external extractor or the burner. Operating in conditions that eliminate the need for separation devices can minimize the capital expenditure for the practice of ISPR liquid-liquid extraction. In addition, by minimizing the size of the extractors by maximizing the interfacial area between the phases of the fermentation broth and the extractant for a given set of physical properties of the fermentation broth and the extractant, the ability to effect phase separation can be maintained. of the broth fermentation and extractant economically. By eliminating the capital and operating cost of separation devices such as centrifuges, the net present value of a plant producing dry and ground corn to an alcoholic product using an ISPR liquid-liquid extraction process can be improved.

In another embodiment of the processes and systems described in the present disclosure, the design of the extractor including the phase separation capability can be adapted to support the physical properties of the fermentation broth and the extractant. If undissolved solids are not removed from the raw material suspension, or if the concentration of the alcoholic product in the fermentation broth is too low, it may not be possible to remove the sufficient amount of alcoholic product to maintain the productivity of a plant that uses an extractor that does not include phase separation equipment. Therefore, the present invention provides processes and systems that include the removal of solids, as well as the recovery of the alcoholic product through the use of an external extractor, where the extractor has been designed to improve the phase separation capacity for the maximum recovery of the alcoholic product.

An illustrative process of the present invention is described in Figure 8. Some processes and flows in Figure 8 have been identified with the same name and numbering used in Figures 1-7, and represent the same processes and currents or processes and flows similar to those described in Figures 1-7.

The raw material 12 can be processed and the solids can be separated (100) as described in the present description with reference to Figures 1-7. Briefly, the raw material 12 can be liquefied to generate the suspension of raw material, which comprises undissolved solids, fermentable sugars (or fermentable carbon source) and, depending on the raw material, oil. Then, the raw material suspension can be subjected to separation methods to remove the solids in suspension and generate a wet cake, an aqueous solution 22 (or centered) comprising dissolved fermentable sugars and, optionally, an oily stream. The separation of solids can be achieved by a series of means including, but not limited to, centrifugation in bowl decanter, three-phase centrifugation, disk-spin centrifugation, filtration centrifugation, decanter centrifugation, filtration, vacuum filtration. , band filter, pressure filtration, membrane filtration, microfiltration, filtration through the use of a screen, sieve separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, settling gravity, vortex separator or combinations of these.

The aqueous solution 22 and the microorganism 32 can added to the fermentation 30, wherein the fermentable sugars are fermented by the microorganism 32 to produce the stream 105 comprising the alcoholic product. In some embodiments, during fermentation, a portion of the stream 105 can be transferred to the extractor 120 (or withdrawal 120), where the stream 105 comes into contact with the extractant 124. In some embodiments, the extractant can be stored in a tank. or extractant storage unit. In some embodiments, the stream 105 can be removed from the fermentation 30 when the concentration of the alcoholic product and / or other metabolic products reaches a predetermined concentration. In some embodiments, the predetermined concentration may be a concentration of the alcoholic product and / or other metabolic products that adversely affects the metabolism of the microorganism. In some embodiments, stream 105 can be removed from fermentation 30 when fermentation is initiated. In some embodiments, the stream 105 may be removed from the fermentation 30 to minimize the effects of the alcoholic product in the microorganism 32. In some embodiments, the fermentation 30 may comprise one, two, three, four, five, six, seven, eight or more termendores.

In some embodiments, extractant may be added to the fermentation 30. In some embodiments, a portion of the broth of fermentation comprising extractant can be transferred to the extractor 120 and, in some embodiments, the extractant can be recovered from the fermentation broth comprising the extractant. By adding extractant to fermentation 30, ISPR can be initiated in fermentation 30.

The alcoholic product or a portion thereof, the transfers of stream 105 to extractant 124 and stream 122 comprising extractant enriched in alcoholic product can be conducted to separation 130. Stream 127 comprising the most depleted fermentation broth in alcoholic product may return to fermentation 30. Separation 130 removes a portion of the alcoholic product from stream 122, and stream 125 comprising more depleted extractant may return to extractor 120. In some embodiments, extractor 120 may be external to fermentation. In some embodiments, the fermentation 30 may comprise an extractor. In some embodiments, the extractant, the fermentation broth or both may be at least partially immiscible. The stream 135 can be conducted downstream for further processing (eg, distillation) which includes recovery of the alcoholic product.

During the course of extraction, there may be a loss of extractant or a portion of the extractant. In some embodiments, extractant 124 can be replenished by adding extractant to the extractor 120 or an extractant storage unit. In some embodiments, for example, wherein the extractant can be derived from the raw material or raw material in suspension, the extractant 124 can be replaced by the conversion of the raw material oil or suspension of raw material into extractant. For example, a catalyst may be added to the fermentation 30 to convert the oil of the aqueous solution 22 into fatty acids and / or fatty esters (see, for example, Figure 7D) and a portion of the stream 105 comprising the alcoholic product. , fatty acids and / or fatty esters can be transferred to the extractor 120, wherein the stream 105 can be contacted with the extractant 124. The stream 122 comprising the extractant enriched in alcohol product, fatty acids and / or fatty esters can be conducted separation 130 to generate stream 125, comprising more depleted extractant, fatty acids and / or fatty esters. In some embodiments, stream 125 may be subjected to further processing before returning to extractor 120. For example, fatty esters present in stream 125 may be subjected to hydrolysis to generate a stream comprising alcoholic product and fatty acids. This stream, which comprises alcoholic product and fatty acids can be conducted to extractor 120, or this stream can be combined with stream 122, and the combined stream can be conducted to the separation 130. In another embodiment, this stream comprising alcoholic product and fatty acids can be conducted to the separation 130, or this stream can be conducted to another separation unit to generate a stream of alcoholic product and a stream of fatty acids. The fatty acid stream can be conducted to the extractor 120 and the alcohol product stream can be combined with the stream 135 and subjected to further processing for the recovery of the alcoholic product.

In some embodiments, the phase separation of the fermentation broth and the extractant after passing through an extractor may be insufficient, so that an unacceptable level of extractant dispersed in the fermentation broth that returns to the fermenter remains, and / or remains an unacceptable level of droplets of the fermentation broth in the extractant advancing towards distillation. In some embodiments, the phase separation of the fermentation broth and the extractant can be improved by processing a heterogeneous mixture leaving the top or bottom of an extractor by one or more hydrocyclones or similar vortex devices. In some embodiments, instead of an extractor, a static mixer can be used to contact the fermentation broth and the extractant, and the heterogeneous mixture formed can be pumped through of one or more hydrocyclones or similar vortex devices to effect a separation of the aqueous (eg, fermentation broth) and organic (eg, extractant) phases. In some embodiments, one or more hydrocyclones or similar vortex devices may be used to remove liquid or droplets of liquid from a gas stream. In some embodiments, the gas stream may come from the thermistor. In some embodiments, the gas stream may come from a degassing device.

In a batch or semi-batch fermentation process, when a portion of the fermentable sugars has been metabolized by the microorganism 32, the stream 103 comprising the beer can be conducted downstream to the separation 140 to separate the alcoholic product from the beer. The stream 145 comprising the alcoholic product can be conducted downstream for further processing (eg, distillation), which includes the recovery of the alcoholic product. In a continuous fermentation process, the stream 103 comprising the beer can be conducted downstream to the separation 140 to separate the alcoholic product from the beer. The stream 142 comprising coarse vinasse can be conducted downstream for further processing, which includes the removal of solids and the generation of light vinasse.

In some embodiments, fermentation 30 may comprising two or more fermentors, and the stream 105 may comprise multiple streams combined, from the two or more fermenters. In some embodiments, the combined multiple streams may be conducted to the extractor 120. In some embodiments, the stream 127 may be divided, and the stream portions 127 may return to the multiple bulkers. In some embodiments, the extractor 120 can be a series of units connected together, in parallel or in series.

In some modalities, the extraction can be done for a certain period of time. The extraction can be carried out, for example, until the concentration of the alcoholic product in the fermentation 30 is low enough so that the separation 140 is not required. In some embodiments, the extraction can be carried out for a prolonged period of time.

In some embodiments of the processes and systems described in the present description a decanter may be used for phase separation. In some embodiments, a decanter may be used in combination with an extractor. In some embodiments, decanter surfaces can be modified to improve phase separation. For example, the decanter surfaces can be modified by the addition of hydrophilic and / or hydrophobic surfaces.

In some modalities, oxygen, air may be added and / or nutrients to stream 125 and / or stream 127. In some embodiments, nutrients may be soluble in the extractant. In some embodiments, the oxygen concentration can be measured in the various streams, and can be used as part of a control circuit to vary the oxygen flow in the process. In some embodiments, temper may be added to extractor 120 to allow more effective titles. In some embodiments, the separation 130 and 140 may be extractors. In some embodiments, these extractors may use water to extract the alcohol product from the extractant, and the alcoholic product may subsequently be separated from an aqueous phase. In some embodiments, the extractant can be infused with solutes that improve its ability to extract the alcoholic product from the fermentation broth. In some embodiments, a compensation tank may be located between the extractor 120 and the separation 130, as a means for balancing the concentration of alcohol product in the extractant before separation (eg, distillation).

In some embodiments, the extractor 120 can be designed to use the CO2 generated during fermentation in the mixing of the fermentation broth and the extractant. In some embodiments, the extractor 120 can be designed to allow immediate decoupling of the CO2 from the fermentation broth. This design would facilitate control of level of mixing by the CO2 bubbles that are released in the extractor 120. In some embodiments, the fermentation broth can be removed from the fermentation 30 to minimize the concentration of CO2 in stream 105. In some embodiments, the design of the zones The decoupling of the extractor may include surfaces to promote phase separation between the fermentation broth and the extractant. In some embodiments, hydrophilic and / or hydrophobic surfaces may be installed in the decoupling zones to improve phase separation. In some modalities, the external extractor may include internal components or output ports for CO2. For example, a coalescence filter can be added to the external extractor.

In some embodiments to minimize mixing with the CO2 the extractor can be designed with a small diameter in the lower part of the extractor, which widens towards the upper part of the extractor (for example, conical shape). In some embodiments, the extractor can be designed with a gradual increase in diameter. For example, the extractor may comprise a first region of constant diameter followed by a gradual increase in diameter to a second region of constant diameter. In some embodiments, the extractor may also comprise a second gradual increase in diameter to a third constant diameter region. In some embodiments, the extractor may comprise one or more gradual increases in diameter. In some embodiments, the extractor may comprise one or more regions of constant diameter.

During the course of the fermentation, the gas content (eg, CO2) of the fermentation broth changes and these gases can be removed from the fermentation broth by the use of a gas desorber. The amount of gas desorbed from the fermentation broth can be adjusted by varying the flow through the gas desorber and / or the pressure of the gas desorber. In some embodiments, the amount of CO2 in the fermentation broth can be reduced before transferring the fermentation broth to an extractor. For example, CO2 can be desorbed from the fermentation broth by a gas desorber or any means known to those skilled in the art. In some modalities, the removal of CO2 can be done at or below the ambient pressure. In some embodiments, fermentation may continue in the extractor, and CO2 may be produced by the microorganism. In some embodiments to minimize the mixing of the CO2 in the extractor, the residence time of the fermentation broth in the extractor can be reduced. In some embodiments, the residence time can be reduced by changing the height of the extractor. In some modalities, you can reduce the height of the extractor. Reducing the height of the extractor can reduce the number of theoretical extraction stages. In some embodiments, to maintain the number of theoretical extraction stages, the extractor can be replaced with two or more extractors of reduced height. In some embodiments, the two or more extractors may be in series. In some embodiments, the two or more extractors may be connected. In some embodiments, the two or more extractors may be connected to maintain the flow in countercurrent. In some embodiments, a degassing step may be added to one or more extraction stages.

With reference to Figure 8, in some embodiments, the droplet size of the dispersed phase of the extractor 120 can be measured and adjusted by various means to improve the mass transfer rate. For example, droplet size can be measured by the use of particle size analysis, such as focused beam reflectance measurement (FBRM®) or particle display and measurement technologies (PVM®) (Mettler-Toledo, LLC, Columbus OH). In some embodiments, the fermentation broth may be the dispersed phase and the extractant may be the continuous phase, and under these conditions, the solids present in the fermentation broth may interact to a lesser degree with the extractant. In some embodiments, the conditions of separation 130 can be controlled to minimize the effects of oxidative and thermal instability in the extractant.

In some embodiments, the quality of the extractant can be monitored and the extractant can be replenished at a frequency necessary for the successful production of the alcoholic product. In some embodiments, the extractant can be absorbed by the solids of the coarse vinasse. The coarse vinasse can be separated into streams of liquid (eg, light vinasse) and solid, and the solids can be washed to recover the extractant. In some embodiments, the temperature of the extractor 120 can be adjusted to improve the efficiency of the total process. In some embodiments, the fermentation broth and extractant streams to the extractor 120 may be co-current or countercurrent. In some embodiments, membranes may be used to minimize the mixing of the fermentation broth and the extractant. In some embodiments, the extractant may be polymer pellets or inorganic pellets that absorb the alcoholic product. In some embodiments, the polymer pellets or inorganic pellets can, preferably, absorb the alcoholic product.

In some modalities, measurements such as within line, in line, at the foot of the line or in real time can be used to measure the concentration of alcoholic product and / or metabolic byproducts in the various streams. These measurements can be used as part of a control circuit to vary the flow between the various units or vessels (eg, fermentation 30, extractor 120, separations 130 and 140, etc.) and to improve the overall process.

In Figure 9 another illustrative process of the present invention is described. Some processes and flows of Figure 9 have been identified with the same name and numbering used in Figures 1-8 and represent the same processes and flows or processes and flows similar to those described in Figures 1-8.

The raw material 12 can be processed and the solids can be separated (100) as described in the present description with reference to Figures 1-7. In some embodiments, raw material 12 can be mixed with recirculated water (eg, stream 162) generated by evaporation 160. As described in the present disclosure, raw material suspension can be subjected to separation methods to remove solids in suspension and generate a wet cake 24, an aqueous solution 22 (or centered) comprising dissolved fermentable sugars and, depending on the raw material, oil. The wet cake 24 can be dried in dryer 170 and used to produce DDGS. In some embodiments, the wet cake 24 can be resuspended with water (eg, recycled water / stream 162) and subjected to separation to remove other fermentable sugars and generate a wet cake washed (eg, 74, 74 ', as described in Figures 4 and 5). In some embodiments, the wet cake streams 24, 74 and 741 may be combined, and the combined wet cake streams may be dried in a dryer 170 and used to produce DDGS.

The aqueous solution 22 and the microorganism 32 can be added to the fermentation 30, wherein the fermentable sugars are metabolized by the microorganism 32 to produce the stream 105 comprising the alcoholic product. In some embodiments, enzymes may be added to the fermentation 30. The stream 105 may be conducted to the extractor 120 and may be contacted with the extractant 124. The stream 127, which comprises the most depleted fermentation broth in alcoholic product, may return to the fermentation 30, and stream 122, which comprises the extractant most enriched in alcoholic product, can be conducted to separation 130. In some embodiments, extractor 120 can operate so that stream 122 contains a minimum cell mass and a minimum substrate. The separation 130 can damage the microorganism 32 or the substrate, which results in a decrease in the fermentation rate. By operating the extractor 120 with a minimum cell mass and substrate, the potential damage due to separation 130 can be minimized. The stream 125 comprising the most depleted extractant can return to the extractor 120. The stream 135 from the separation 130 purification 150 can be conducted for further processing, which includes the recovery of the alcoholic product. In some embodiments, extractant may be added to the fermentation 30. In some embodiments, a portion of the fermentation broth comprising extractant may be transferred to the extractor 120 and, in some embodiments, the extractant may be recovered from the fermentation broth comprising the extractant. In some embodiments, the flow rates of the fermentation broth and extractant to the extractor can be modified to improve phase separation. For example, lower total flows entering the extractor in the early or late stages of fermentation may improve phase separation of the fermentation broth and the extractant.

As described in the present description, after a batch fermentation process or as a constant stream of effluent in a continuous fermentation process, the stream 103 comprising the beer can be conducted downstream to the separation 140 to separate the alcoholic product of coarse vinasse 142. The use of an upstream solids removal process can reduce the content of undissolved solids in the fine temper and, therefore, centrifugation of coarse vinasse 142 may not be required to remove the solids. Therefore, the coarse vinasse 142 can be conducted directly to the evaporation 160. The syrup 165 generated by the evaporation 160 can be mixed with the wet cake 24, 74, 74 'in dryer 170 to form DDGS.

In some embodiments, the countercurrent water comprising total suspended solids (TSS) from the coarse vinasse can be used (or recielated) to prepare the raw material suspension. In some embodiments, the coarse vinasse, or a portion of the coarse vinasse, can be processed by means of a solid separation system including, but not limited to, turbo filtration or ultracentrifugation before evaporation, or coarse vinasse, or a portion of the coarse vinasse, can be processed for the purification of the self-cleaning water.

In some embodiments where thicker grain solids are removed from the liquefied temper, the coarse vinasse produced may contain fine solids and insoluble fragments of microorganisms, and these dispersed solids may be removed by turbo filtration. The turbo filtration may include subjecting a feed suspension to centrifugal movement by means of a sieve that can retain the fine solids. When formed in a wet cake, these fine solids may contain a portion of extractant absorbed on the surface and inside the pores of the fine-grained particles. In some cases, washing the wet cake with water is insufficient to recover the extractant from the wet cake. In some embodiments, a concentrated stream of alcoholic product, such as the organic phase, to recover extractant from the wet cake of coarse vinasse. In some embodiments, this organic phase can be formed in a decanter. In some embodiments, the wet cake washed with the alcoholic product can then be washed with water to recover the alcoholic product from the wet cake.

In some embodiments, the processes and systems described in the present disclosure may include a deposit (or tank or container) of extractant. The extractant can be added to the extractant reservoir and this extractant can be circulated to an extractor. In some embodiments, the extractant may be conducted to an extractor and an extractant stream may return to the extractant reservoir. In some embodiments, the extractant from an extractant reservoir can be circulated to an extractor and / or fermentor. In some embodiments, an extractant stream may be circulated between an extractant reservoir, an extractor and a fermenter. In some embodiments, at the end of the fermentation, the content of the extractant deposit, the extractor and / or the fermenter can be subjected to further processing to recover the alcoholic product.

The separation or extraction of the alcohol product from the extractant can be carried out by using methods known in the art, including, but not limited to, siphoning, decanting, centrifugation, gravity settler, phase separation assisted by membrane, and the like. In some embodiments, extraction can be accomplished by the use of, for example, mixer-settlers. The mixer-settlers are stage extractors and are available with various elements for mixing, such as pumps, agitators, static mixers, mixing "T", impact devices, circulation screens or rain contact buckets. Examples of mixer-settlers are shown in Figures 10A-10H. For example, Figure 10A illustrates a mixer-settler that uses a pump as the mixing source. Figure 10B illustrates a mixer-settler that uses a mixer as a mixing source. Figure 10C illustrates a mixer-settler that uses a static mixer as a mixing source. Figure 10D illustrates a mixer-settler using a mixing "T" as the mixing source. Figure 10E illustrates a mixer-settler that uses an impact mixer as a mixing source. Figure 10F illustrates a mixer-settler that uses a rain or mesh sieve as a mixing source. Figure 10G illustrates a mixer-settler that uses a centrifuge as a settler. Figure 10H illustrates a mixer-settler that uses a hydrocielon or vortex separator as a settler. In some embodiments, one or more mixing devices may be used in the processes and systems described in the present disclosure.

In some embodiments, the mixers may comprise agitators such as, for example, flat blades, turbines seated in blades or curved blades. The droplet size produced by agitated mixers can be controlled by the design of the agitator, the tank design, the agitator speed and the mode of operation. In static mixers, the droplet size can be controlled by the mixer diameter and the flow rate. For example, the droplet size can be controlled by varying the flow in the mixer during the course of fermentation. In some embodiments, gases and mixers may be used for mixing purposes.

In some embodiments, one or more mixer-settlers may be used in the processes and systems described in the present disclosure. In some embodiments, the one or more mixer-settlers may be arranged in series or in countercurrent mode, as illustrated in Figures 101 and 10J. In some embodiments, the mixer-settlers can be stacked in a column arrangement, which provides multiple mixing and settling zones. In some embodiments, the settler may comprise hydrophilic or hydrophobic surfaces to promote phase separation.

In another embodiment, column extractors may be used in the processes and systems described in the present description.

Centrifugal extractors. The column extractors are differential extractors that provide the conditions for mass transfer along their entire length with a constantly changing concentration profile. Different types of differential extractors can be divided into non-mechanical, pulsed and stirred by rotation. Centrifugal extractors are a separate class of differential extractors and the Podbielniak® centrifugal contactor is one of those types.

In some embodiments, non-mechanical spray towers may be used in the processes and systems described in the present disclosure. An example of a non-mechanical spray tower includes a non-mechanical spray tower with no internal components in the column. To determine the droplet size, the number of nozzles and the nozzle diameter can be used. In some modalities, the spray tower can have internal components. In some embodiments, a spray tower may comprise a helical pipe. The helical pipe can allow the droplet increase and the additional mixing of the fermentation broth and the extractant. In some embodiments, non-mechanical extractors, such as packed towers, sieve trays and baffle trays, may be used in the processes and systems described in the present disclosure. Examples of these extractors are shown in Figure 10K. In some modalities, the packaging of these extractors can be random or structured.

In some embodiments pulsed extractors may be used in the processes and systems described in the present disclosure. Pulse-pulsed extractors have different designs, and include reciprocating trays or vibrating plates, where the trays move vertically. In addition, the complete packaged column and / or tray of sieves may vibrate vertically to promote smaller droplets in the dispersed phase and higher mass transfer. Examples of these extractors are shown in Figure 10L. In some embodiments, in the processes and systems described in the present description, rotating or rotating disk contactors may be used. Examples of these extractors are shown in Figure 10M.

In some embodiments, agitated extractors may be used in the processes and systems described in the present disclosure. For example, extractors stirred with centrifuges can provide high mass transfer rates and a clear phase separation. In some embodiments, agitated columns may be used in the processes and systems described in the present description. For example, agitated columns with internal components can provide high mass transfer rates.

One aspect of a liquid-liquid extraction process is to determine the successful operating conditions for the extractor during the course of a constantly changing fermentation. For example, a typical batch fermentation of corn to an alcoholic product uses an initial inoculum of microorganism (or cell mass) added to a certain volume of the fermentation broth in the fermenter, followed by additional filling of the fermenter to a specified volume. The fermentation is allowed to proceed until a predetermined amount of the fermentable carbon source (eg, sugar) is consumed. During the course of batch fermentation, cell mass concentrations, reaction intermediates, reaction by-products and substrate components change over time, as do the physical properties of the fermentation broth, which includes viscosity, density and surface tension . To improve the performance parameters of the fermentation, for example, speed, title and performance parameters of the production, as well as the economic aspects of the plant, such as sales volume, return on investment and profit, the extractor can be made operate in a variable manner to overcome the drawback of the changing fermentation broth. In addition, the properties of a dynamic fermentation can affect the size limits of the extractor. The proper integration of the operation of the extractor and the therminator can benefit from the use of mathematical models of the process (see, for example, Daugulis and Kollerup, Biotechnology and Bioengineering 27: 1345-1356, 1985). In addition, it is valuable to increase the mathematical model, for example, by configuring the key parameters of the model with experimental data. The design parameters for differential extractors that must be considered to improve the speed, the title and the yield of the fermentation process include the maximum total flow to the extractor per cross-sectional area of the extractor column, as well as the ratio between the extractant and the extractant. the height of the extractor required to remove the sufficient alcoholic product in a given fermentation broth. During a batch fermentation, the change of the maximum flow per unit area and the height of the extractor may be required. Another consideration for differential extractors is the droplet size of the dispersed phase. The proper droplet size can be an equilibrium between small enough to provide adequate mass transfer, but large enough to allow a clear phase separation exiting the extractor. In stepped extractors, the mixing intensity required for an efficient mass transfer, the corresponding time to sediment and / or the energy needed to separate the phases are additional elements that must be considered. In any type of extractor, by stages or differential, the broth ratio of Fermentation to extractant that is fed to the extractor is important to determine the size of the extractor.

In some modalities, if an extractor of fixed size was used and the maximum admissible flow that prevents the flood to the extractor will vary from a low value to a high value (for example, from 1/3 to 2/3 of the maximum for an exhaust extractor). given design) during the course of the fermentation due to changes in the physical properties and fermentation broth concentrations, then, the flows to the extractor can be varied, without exceeding the maximum flow, but completing the fermentation in a reasonable time. In some embodiments, if an extractor is agitated, the agitation rate may be varied during the course of fermentation to compensate for changes in the fermentation broth. The droplet size can be measured inside the extractor, and the speed to maintain a fixed droplet size can be controlled throughout the fermentation to compensate for changes in the fermentation broth. The mass transfer magnitude that occurs at any time point can be evaluated by measuring the concentrations of the alcohol product in the inlet and outlet streams, and adjusting the conditions (eg, flow, flow rate, agitation) to control the mass transfer during the course of fermentation.

In some embodiments, multiple extractors of different sizes may be used and the conditions (e.g., flow, flow rate, agitation) of each extractor may be adjusted to provide better control of the fermentation process. In some embodiments, the proportion of fermentation broth to extractant can be modified to improve extraction efficiency, increase the concentration of alcohol product in the extractant (equivalent to greater efficiency) and reduce the necessary flows through the extractor.

In additional embodiments of the processes and systems described in the present description, there may be two or more fermentation broths or aqueous streams. An extractant phase which has absorbed the alcoholic product of a first aqueous stream can be contacted with a second aqueous stream containing less alcoholic product than the first aqueous stream or fermentation broth, which allows the transfer of the alcoholic product from the phase enriched in extractant to the second aqueous phase. In some embodiments, contacting the enriched extractant with a dilute aqueous stream can take place in a multi-stage contact device or in a static mixer, followed by a settler. In some embodiments, the contacting of the enriched extractant with a dilute aqueous stream can take place in the same device in where the depleted extractant is put in contact with the fermentation broth. An extractor with perforated deflectors would allow the downward flow of the fermentation broth and a dilute aqueous stream in separate compartments, while an extractant depleted in alcoholic product can form a continuous phase in all the compartments. An advantage of this configuration is that, if the extractant remains confined to the closed volume of an extractor, a reduced amount of extractant would be needed in the production plant. Another advantage of this configuration is that the extractant is not subject to possible degradation during the distillation and, therefore, may exhibit a longer shelf life. By transferring the alcoholic product to a homogeneous aqueous stream, the alcoholic product can be conducted to more than one desorbing column by dividing the diluted aqueous stream, taking into account the capacities of the column and the integration of heat. The need to clean equipment exposed to an extractant can be reduced when the alcoholic product is extracted in an aqueous medium, during or immediately after the alcoholic product is extracted from the fermentation broth.

In some embodiments, the alcoholic product can be transferred from the fermentation broth to a second aqueous stream or an extractant by a selective barrier for the transport of the alcoholic product. In some modalities, this barrier may be provided by a membrane material. The membrane material may be organic or inorganic. Examples of membrane material include polymers and ceramics. In some embodiments, the alcoholic product can be separated from the fermentation broth by the use of a hydrogel. In some embodiments, the hydrogel may comprise functional elements that promote interaction with an alcoholic product, such as, but not limited to, hydroxyl functionality, hydrocarbon character, network size, and the like. In some embodiments, a hydrogel may comprise a polymeric network structure or polymeric formulations. Examples of polymeric formulations include, but are not limited to, one or more of the following: acrylic acid, sodium acrylate, hydroxyethyl acrylate, methacrylate, hydroxybutyl acrylate, butyl acrylate, vinyl polyethylene oxide, vinyl polypropylene oxide , vinyl polytetramethylene oxide, acrylates and diacrylates of polyglycols, polyvinyl alcohol and polyvinyl alcohol derived from hydrocarbons, and styrene and styrene derivatives. In some embodiments, the hydrogel may comprise hydroxyethyl acrylate and methacrylate, hydroxybutyl acrylate and methacrylate or butyl acrylate and methacrylate.

In other embodiments of the processes and systems described in the present description, the fermentation broth can be removed from the bottom of the higher pressure burner that the atmospheric and go through a first tank of instantaneous vaporization that works at atmospheric pressure to release dissolved gases, such as CO2. This first flash tank can be a degassing hood and the vapors of the first flash tank can be combined with the vapors of the fermenter and directed to a scrubber. In some embodiments, the fermentation broth from the first flash tank can be passed through a second flash tank operating at a pressure lower than atmospheric to release more dissolved gases, such as CO2. This second flash tank can be a degassing cyclone, and the vapors from this second flash tank can be recompressed to atmospheric pressure, cooled and partially condensed before being combined with vapors from the fermenter and directed to a scrubber. The fermentation broth leaving this second flash tank can be pumped to an extraction column that operates at a pressure higher than atmospheric, so that the remaining or newly formed dissolved gases do not lead to the formation of a vapor phase in the extraction column.

In another embodiment of the processes and systems described in the present description, the fermentation broth can be conducted to an extractor and contacted with the extractant to generate an aqueous stream and an organic stream, comprising the extractant and the alcoholic product. This organic stream can be conducted to a flash tank (for example, vacuum venting) to separate the alcohol product from the extractant. In some embodiments, the extractant stream from the flash tank can be recielated to the extractor and / or the thermistor. In some embodiments, the organic stream can be conducted to a second extractor before the flash tank. This second extractor can be used to remove, for example, wastewater from the organic stream. The extractors can be siphons, decanters, centrifuges, gravity settlers, mixer-settlers, or combinations of these. In some embodiments, the extractant may be an oil such as, but not limited to, tallow, corn, canola, capric / caprylic triglycerides, castor, coconut, cottonseed, fish, jojoba, lard, flaxseed, cow's foot , oiticica, palm, peanut, rapeseed, rice, safflower, soy, sunflower, tung, Mexican pinion and mixtures of vegetable oil, or fatty acids derived from these.

In some embodiments of the processes and systems described in the present description, automatic self-cleaning filtration can be used in these processes and systems. The fermentation broth can be removed from a It can be cooled and cooled by the use of a cooler (for example, an existing cooler in a fermentation production facility) before entering an automatic self-cleaning filter. As the clarified temper passes through the filter, some particulates may be retained in the filter screen. When a portion of the clarified temper flows back to the screen carrying the particulates with it, other filters can be simultaneously subjected to reflux to discharge a concentrated stream of solids. In some embodiments, a portion of the clarified temper may enter the top of an extractor, while an extractant is introduced into the lower part of the extractor. The clarified temper and the extractant can be passively contacted by differences in density, or with the aid of mechanical movement (eg, a Karr® column) by means commonly used in the art. In some embodiments, a stream of extractant organic liquid containing the alcoholic product emerges from the top of the extractor and a stream of aqueous liquid from the fermentation broth that has been at least partially depleted in alcoholic product, with respect to the clarified temper , emerges from the bottom of the extractor. The aqueous liquid stream and the stream of concentrated solids can be combined and returned to the fermenter. The extractant stream enriched in alcoholic product it can be heated in a heat exchanger that transfers heat from an extractant stream depleted in alcoholic product, which originates in the lower part of the extractor. After releasing a little heat, the depleted extractant can be further cooled with water in a heat exchanger to reach a temperature suitable for fermentation. The circulation of the fermentation broth can include a path through a heat transfer device and a mass transfer device, which allows the removal of heat and alcohol product each time it passes through an external cooling circuit. In addition, in some embodiments, the heat and alcohol product removal rate can be balanced with the production rate of heat and alcohol product during fermentation by adjusting the circulation flow through the external cooling circuit, adjusting the fluid flow of cooling in a heat exchanger and / or adjusting the flow of extractant.

In some embodiments of the processes and systems described in the present disclosure, phase separation between extractant and fermentation broth can be improved by changing the temperature and / or the pH of the process. For example, the process may be operated at temperatures and / or pH different from the temperature and / or pH of the thermistor. In some modalities, the process can be operated at a Reduced pH compared to the fermentor. In some embodiments, the process can be operated at a higher temperature compared to the fermentor. In some embodiments, the process can be operated at a reduced pH and a higher temperature compared to the fermentor. A higher temperature may increase the mass transfer kinetics of the alcoholic product between the aqueous and organic phases, and may increase the kinetics of coalescence of the extractant droplets dispersed in the aqueous phase and the aqueous droplets dispersed in the organic phase. In some embodiments, the temperature inside an extractor containing fermentation broth and extractant can be increased by heating the fermentation broth and / or extractant entering the extractor. The fermentation broth can be heated directly by the injection of steam or, indirectly, by means of a heat exchanger. In some embodiments, the extractant that feeds the extractor may originate by distillation, where its temperature may already be elevated. In some embodiments, the extractant can be cooled to a temperature higher than the fermentation temperature.

In some embodiments, a reduced pH can minimize the solubility and dispersibility of the extractant in the aqueous phase of the broth. In some embodiments, the extractant may a fatty acid with a known value of pKa associated In some embodiments, the pH of the fermentation broth can be reduced to below the pKa of the extractant, such that the carboxylic acid groups of the fatty acid are substantially protonated. In some embodiments, the pH can be reduced by introducing gaseous CO2 into the fermentation broth, or by injecting a small amount of liquid acids, such as sulfuric acid or any other organic or inorganic acid, into the fermentation broth. In some embodiments, the pH of the fermentation broth after separation of the extractant can be adjusted to the pH of the fermentation.

In some embodiments in which the phase of the extractant is the continuous phase, the aqueous phase may be distributed or dispersed in the phase of the extractant. For example, the fermentation broth comprising the alcoholic product can be conducted to an extractor (for example, external extractor) by means of a distributor or a dispersion device. In some embodiments, the dispensing or dispersing device may be a nozzle, such as a spray nozzle. In some embodiments, the distribution or dispersion device may be a spray tower. As an example, droplets of the fermentation broth can be passed through the extractant and the alcoholic product transferred to the extractant. The droplets of the fermentation broth coalesce in the lower part of the extractor and can return to the fermentor The extractant comprising the alcoholic product can be subjected to further processing to recover the alcoholic product as described in the present description. In addition, at the end of the fermentation, the residual alcohol product in the fermenter can also be subjected to further processing to recover the alcoholic product. In some embodiments, the phase of the extractant may be in countercurrent.

In some embodiments in which the phase of the extractant is the continuous phase and the aqueous phase is the dispersed phase, the mass transfer rates can be improved by the use of electrostatic spraying to disperse the aqueous phase in the phase of the extractant. In some embodiments, one or more spray nozzles may be used for electrostatic spraying. In some embodiments, the one or more spray nozzles may be an anode. In some embodiments, the one or more spray nozzles may be a cathode.

In some embodiments, the effluent from the extractor may be used to improve phase separation. For example, a portion of enriched extractant (ie, extractant enriched in alcohol product) from the top of the extractor may return to the top of the extractor as reflux, and the remaining enriched extractant may be subjected to further processing to recover the product. alcoholic. To Addeemmááss, a portion of broth from Depleted fermentation from the lower part of the extractor can return to the lower part of the extractor as reflux, and the remaining depleted fermentation broth can return to the fermenter. In another embodiment, the enriched extractant can leave the top of the extractor towards a decanter and separate into a heavy phase and a light phase. The heavy phase from the decanter can be conducted to the top of the extractor to improve phase separation. The light phase from the decanter can be subjected to further processing to recover the alcoholic product.

In some embodiments of the processes and systems described in the present disclosure, a multiple-pass extractant stream may be used to recover the alcoholic product. For example, multiple headers and extractors can be used, where the fermentation cycle of each fermenter takes place at a different time point. With reference to Figure 11A as an example, the fermenter 300 is at an earlier time point compared to the fermenter 400, which is at an earlier time point compared to the fermenter 500. The fermentation broth comprising the alcoholic product 302 from of the fermenter 300 can be contacted with the extractant 307 in the extractor 305, and the alcoholic product can be transferred to the extractant to generate the extractant enriched in alcoholic product 309. Extractant enriched in alcoholic product 305 can be conducted to extractor 405. The fermentation broth comprising alcoholic product 402 from termenter 400 can be conducted to extractor 405 to produce the extractant enriched in alcoholic product 409. The extractant enriched in alcoholic product 409 can be conducted to extractor 505. The fermentation broth comprising alcoholic product 502 from fermenter 500 can be conducted to extractor 505 Extractant enriched in alcoholic product 509 from extractor 505 can be processed to recover the alcoholic product. The fermentation broth depleted in alcoholic product (304, 404, 504) can return to the fermenters 300, 400 and 500, respectively. The number of heaters and extractors may vary depending on the operating establishment. A benefit of this process is, for example, the reduction in the total extractant processing and the extractor size.

In another embodiment of this example, there may be an additional fermenter F 'and an additional extractor E' (Figure 11B). In this embodiment, when the fermenter 500 (which is at a later time point compared to the fermenters 300 and 400) completed the fermentation, the fermenter 500 can be taken offline and, in some embodiments, the fermenter 500 can be subjected to sanitation and / or sterilization procedures, such as plant cleaning procedures (CIP) and in-plant sterilization (SIP, for its acronym in English). When the heater 500 goes offline, the heater F 'can be brought online. In this mode, the heater F 'is at a previous time point compared to the heater 300, which is at a previous time point compared to the heater 400. Like the description of Figure 11A, the fermentation broth which comprises the alcoholic product F'-02 from the fermenter F 'can be contacted with the extractant in the extractor E1, and the alcoholic product can be transferred to the extractant to generate an extractant enriched in alcoholic product E'-09. The extractant enriched in alcoholic product E'-09 from the extractor E 'can be fed to the extractor 305. The fermentation broth comprising the alcoholic product 302 from the fermenter 300 can be conducted to the extractor 305 to produce the extractant enriched in alcoholic product 309. The extractant enriched in alcohol product 309 can be conducted to extractor 405. The fermentation broth comprising the alcoholic product 402 from the fermenter 400 can be conducted to the extractor 405. The extractant enriched in alcoholic product 409 from the extractor 405 can process to recover the alcoholic product. The fermentation broth depleted in alcoholic product (F'-04, 304, 404) can return to the fermenters F ', 300 and 400 respectively. In some embodiments, this process may be repeated for multiple cycles, for example, at least one, at least two, at least three, at least four, at least five, at least ten, at least fifteen, at least twenty or more cycles. In some modalities, the process of putting off-line equipment and placing additional equipment online can be manual or automated. A benefit of this process is the reduced flow of the extractor to recover the product (for example, distillation).

In some embodiments, an extractant can reduce the flash point (i.e., flammability) of the alcoholic product. Flash point refers to the lowest temperature at which the spread of the flame through the surface of a liquid takes place. The flash point can be measured, for example, by the use of ASTM D93-02 method ("Standard Test Methods for Flash Point by Pensky-Martens Closed Tester'j.The reduction of the flash point of the alcoholic product can improve the conditions of safety of an alcohol producing plant, for example, by minimizing the risk of fire of the potentially flammable alcoholic product.When improving safety conditions, the risk of injury minimizes, as well as the risk of material damage and loss of income. In some embodiments where the inactivation of the microorganism is required, an extractant may improve the inactivation of the microorganism.

In some embodiments, the processes described in the present description can be integrated extraction-fermentation processes by the use of online measurements, in-line, on-line and / or in real-time, for example, concentrations and other physical properties of the fermentation broth and the extractant. These measurements can be used, for example, in feedback loops to adjust and control the conditions of the fermentation and / or the conditions of the extractor. In some embodiments, the concentration of the alcoholic product and / or other metabolites and substrates of the fermentation broth can be measured with the use of any suitable measuring device for in-line measurements, in-line, on-line and / or in real time . In some embodiments, the measurement device may be one or more of the following: Fourier transform infrared spectroscopy (FTIR), near infrared spectroscopy (NIR), Raman spectroscopy, high pressure liquid chromatography (HPLC), viscometer, densitometer, tensiometer, droplet size analyzer, pH meter, dissolved oxygen (DO) probe, and the like. In some modalities, the venting of waste gas from the boiler can be analyzed, example, by means of an in-line mass spectrometer. The measurement of the residual gas vent of the thermidor can be used as a means to identify species present in the fermentation reaction. The concentration of the alcoholic product and other metabolites and substrates dissolved in the extractant can also be measured by the use of the techniques and devices described in the present description.

In some embodiments, the measured inputs can be sent to a controller and / or a control system, and the conditions within the fermenter (temperature, pH, nutrients, concentration of enzymes and / or substrate) can be varied to maintain a concentration, profile of concentration and / or conditions within the extractor (fermentation broth flow, fermentation broth flow to extractant flow, agitation speed, droplet size, temperature, pH, DO content). Similarly, the conditions within the extractor can be varied to maintain a concentration and / or concentration profile within the fermenter. With the use of a control system of this type the process parameters can be maintained, in such a way as to improve the overall productivity of the plant and the economic goals. In some embodiments, real-time control of the fermentation can be achieved by varying the concentrations of the components (eg, sugars, enzymes, nutrients and the like) in the fermenter, variation of the conditions inside the extractor or both.

As an example of an isobutanol fermentation process, the efficiency of the isobutanol extraction in a Karr® column continuously changes as the concentrations of starch, sugars and isobutanol in the fermentation broth change. To maximize the efficiency of the extractor, it may be advantageous to alter the rate at which the isobutanol is removed from the fermentation broth to match the production profile of the isobutanol fermentation. The isobutanol concentrations in the extractant can be maximized, which translates into more energy efficient distillation operations.

As part of a process control strategy, the real-time measurements of isobutanol in the fermentation broth (eg, column feed) can be coupled with the real-time measurements of isobutanol in the extractant and in the fermentation broth impoverished. These measurements can be used to adjust the proportion of fermentation broth to extractant (flows) to the extractor. The flexibility to match the extraction rate of isobutanol with the rate of isobutanol generation can allow the extractor to operate efficiently throughout the extraction. In addition, by maintaining a high concentration of isobutanol in the extractant, the volumetric flow to the distillation columns can minimized, which translates into energy savings for distillation operations. Phase separation can also be monitored with the use of real-time measurements, for example, by monitoring the phase separation rate, the extractant droplet size and / or the composition of the fermentation broth. In some embodiments, phase separation can be monitored by conductivity measurements, dielectric measurements, viscoelastic measurements or ultrasonic measurements. In some embodiments, an automated phase separation detection system can be used to monitor phase separation. This automated system can be used to adjust the flow rates of the fermentation broth and the extractant to and from the extractor and / or adjust the droplet size of the extractant, for example, after mixing the fermentation broth and the extractant. With the use of these real-time monitoring systems, a clear phase separation of the aqueous and organic phases can be achieved.

As another example of the process control strategy, the droplet size can be measured by the use of particle size analysis, such as a process particle analyzer (JM Canty, Inc., Buffalo, NY), reflectance measurement with focused beam (FBRM®) or particle measurement and visualization technologies (PVM®) (Mettler-Toledo, LLC, Columbus OH). In some modalities, These measurements can be in situ characterizations of the particle system in real time. By controlling the droplet size in real time, changes in the shape and dimensions of the droplets can be detected and the process steps can be adjusted to modify the droplet size and improve the mass transfer rate. For example, to control the amount of extractant in the fermentation broth the droplet size can be used. After phase separation, a little extractant may be present in the fermentation broth and, in some embodiments in which the fermentation broth is reclimated to the fermenter, the droplet size monitoring would provide a means to minimize the amount of fermentation. extractant in the fermentation broth that returns to the fermenter. If the amount of extractant in the fermentation broth is too high, then phase separation can be improved, for example, by adjusting the droplet size of the extractant in the extractor and / or adjusting the flow rates of the fermentation broth. and from the extractant to the extractor. These adjustments in the process steps can minimize the amount of extractant in the fermentation broth, as well as minimize the amount of extractant in the lightweight vinasse and DDGS.

In one embodiment of this control strategy, the isobutanol in the fermentation broth would not exceed concentration or target value at which the concentration of isobutanol becomes harmful to the micro-organism. The target value of isobutanol in the fermentation broth can be increased or decreased as the fermentation progresses according to the trajectory of the fermentation. For example, in order to modify the proportions of fermentation broth to extractant or the flow rates of the fermentation broth and of the extractant to an extractor, the continuous comparison of the concentration of isobutanol in the fermentation broth with respect to an objective concentration of isobutanol can be used. To monitor the concentrations of isobutanol in the fermentation broth, in-situ measurements of the fermentation broth can be made by using Fourier transform infrared spectroscopy (FTIR), near-infrared spectroscopy (NIR, by its acronym in English) and / or Raman spectroscopy. In addition, measurements of the thermistor's empty space can be made by using FTIR, Raman spectroscopy and / or mass spectrometry.

In some embodiments, the efficient operation of the extractor may take place in the vicinity of the flood point of the extractor. The use of real-time process control that uses concentration data of the input and output currents can allow the extractor to operate reliably in the vicinity of the point of flood. In some embodiments, extractant monitoring can be used in real time to detect the partition of fermentation broth byproducts or contaminants in the extractant. Byproducts such as alcohols, lipids, oils and other fermentation components can reduce the extraction efficiency of the extractant. Numerous process monitoring techniques can be applied for this measurement, including, but not limited to, Fourier transform infrared spectroscopy (FTIR), near infrared spectroscopy (NIR), high performance liquid chromatography (HPLC), and magnetic resonance imaging. nuclear (NMR). The analytical technique selected to monitor the extractant to detect the presence of by-products or contamination may be a technique different from that used for the determination of alcohol in real time. Real-time data can be used to trigger the recovery of a contaminated extractant or purge a contaminated extractant from the process. These techniques, as well as gas chromatography (GC) and supercritical fluid chromatography (SFC) can also be used to monitor the thermal decomposition of the extractant.

With reference to Figure 12, the systems and processes of the present invention can include means for online measurements, within line, on line and / or in real time (the circles represent the measuring devices and the lines dotted represent feedback loops). Figure 12 is similar to Figure 9, except for the addition of measuring devices for in-line, in-line, line-foot and / or real-time measurements and, therefore, will not be described again in detail .

As an example, in-line measurements of aqueous stream 22 can be used to monitor the concentration of fermentable carbon sources (eg, polysaccharides), oil and / or dissolved oxygen. For example, FTIR can be used to monitor the oil dispersion in the aqueous stream 22, and process image shots can be used to monitor the concentration and size of the oil droplets in the aqueous stream 22. In some embodiments, they can be used. online measurements of fermentation 30 to monitor the removal rates of the alcoholic product. Measurements of fermentable carbon sources, dissolved oxygen, alcohol product and byproducts can be used to adjust the rate of removal of the alcoholic product, to maintain a concentration of alcoholic product in the fermentation 30 that is tolerable for the microorganisms. By maintaining an objective concentration of the alcoholic product, the inhibition and toxicity of the product can be minimized.

Online measurements of current 105 and current 122 can be used to operate feedback loops of the process control. For example, you can the alcoholic product concentration of the stream 105 is used to monitor the flow of this stream to the extractor 120; and the alcohol product concentration of stream 122 can be used to monitor the flow rate of this stream to separation 130 and to set the ratio of fermentation to extractant broth. In addition, in-line measurements of stream 105 and stream 122 can be used to establish the mass balance of the alcohol product in real time. Process control feedback loops for extractor 120 and separation 130 can be used to monitor the quality of the phase separation of the extractant and the fermentation broth. For example, in-line measuring devices can be used to detect the equilibrium of the extractant separation and the fermentation broth, and the feed rates of the extractant and the fermentation broth can be adjusted accordingly to improve phase separation. In-line devices, such as optical devices, can be used to detect the presence of an emulsion layer (eg, oil mixture, aqueous solution and solids) in, for example, extractor 120 and the proportion of fermentation broth can be adjusted. to extractant to minimize the formation of an emulsion layer. Online measurements of stream 135 from separation 130 can be used to monitor the presence of fermentation broth in this stream, and the presence of broth from fermentation in stream 135 may indicate poor phase separation. If the concentration of the fermentation broth in stream 135 is greater than a given target value, process changes, such as flow adjustments or adjustments of the proportion of fermentation broth to extractant can be implemented to improve phase separation. In addition, the concentration of alcohol product in stream 135 can be used as a feedback loop of process control to ensure efficient operation of separation 130.

As another example, on-line measurements of the concentration of alcohol product in stream 127 can be used to monitor the efficiency of the extraction and to maintain a concentration of alcohol product in the fermentation 30 that is tolerable for the microorganisms. In addition, current 127 can be monitored to detect the presence of extractant as a means to minimize the amount of extractant that returns to fermentation 30. For example, to monitor the presence of extractant in stream 127, spectroscopic and image-taking techniques can be used. of process. In addition, a certain concentration of extractant in stream 127 can be maintained to improve the efficiency of extraction and phase separation.

In another embodiment, the stream 135 from the separation 130 can be conducted to the purification 150 for the further processing, including recovery of the alcohol product and extractant 152. Extractant 152 can be conducted to extractor 120. In order to monitor stream 152 for contaminants and degradation products, online measurements can be used. Monitoring current 152 minimizes possible contamination of extractor 120 and fermentation 30. If there is an increase in contaminants in stream 152, this stream may be subjected to further processing to remove these contaminants, for example, by absorption or chemical reaction.

During the extraction process, an emulsion layer can form at the interface of the aqueous and organic phases, and the emulsion layer, composed of solids and extractant (for example, extractant droplets), can accumulate and possibly interfere with the phase separation. To mitigate the formation of the emulsion layer, agitation of the aqueous and organic phases can be used. For example, an impeller can be used to disperse the emulsion layer at the aqueous-organic interface. In addition, a fluid flow, such as a recirculation circuit or vibrations / oscillations, can be used to alter the formation of emulsions. Figures 13A and 13B show illustrative processes to mitigate the formation of an emulsion layer. Figure 13A exemplifies the use of a static mixer in combination with a stirring unit downstream of the settler or decanter for the treatment of an emulsion layer, and Figure 13B exemplifies the use of a static mixer in combination with a stirring unit upstream of the settler or settler for the treatment of an emulsion layer. In some embodiments, other devices, such as coalescing agents or sonic agitation, may be used to disperse the emulsion layer. In some embodiments, these devices may be integrated in the settler or decanter.

The processes and systems described in the present description can be carried out by the use of batch, fed batch or continuous batch fermentation. Batch fermentation is a closed system in which the composition of the fermentation broth is established at the beginning of the fermentation and is not subject to artificial alterations during the fermentation process. In some embodiments of the batch fermentation, extractant may be added to the blender. In some embodiments, the volume of extractant may be from about 20% to about 60% of the work volume of the thermistor.

Batch-fed fermentation is a variant of batch fermentation, in which substrates (eg, fermentable sugars) are added in increments during the fermentation process. Batch fed systems are useful when the repression of catabolites can inhibit the metabolism of the microorganism, and where it is desirable to have limited amounts of substrate in the media. In some modalities, substrate and / or nutrient concentrations can be monitored during fermentation. In some embodiments, parameters such as pH, dissolved oxygen and gases (eg, CO2) can be monitored during fermentation. From these measurements, the rate or amount of substrate and / or the addition of nutrients can be determined. In some embodiments, as the level or amount of fermentation broth decreases during fermentation, more temper may be added to the fermenter to maintain the level or amount of fermentation broth, e.g., maintaining the level or amount of the broth. of fermentation at the beginning of the fermentation process. In some embodiments of fed batch fermentation, extractant may be added to the fermenter.

Continuous fermentation is an open system where fermentation broth is continuously added to a fermenter and a quantity of fermentation broth is removed for further processing (eg recovery of the alcoholic product). In some embodiments, the addition and removal of the fermentation broth may be simultaneous. In some embodiments, equal amounts of fermentation broth may be added and removed from the fermenter. In some continuous fermentation modes, extractant may be added to the fermenter. In some modalities, the Extractant volume may be from about 3% to about 50% of the workload of the thermistor. In some embodiments, the volume of extractant may be from about 3% to about 20% of the work volume of the thermistor. In some embodiments, the volume of extractant may be from about 3% to about 10% of the work volume of the thermistor.

In some embodiments of the processes and systems described in the present description, desorption with gases can be used to remove the alcoholic product from the fermentation broth. The desorption with gases can be carried out by providing one or more gases, such as air, nitrogen or carbon dioxide to the fermentation broth and thereby forming a gas phase of the product containing alcohol. For example, desorption with gases can be accomplished by bubbling one or more gases in the fermentation broth. In some embodiments, the gas may be provided by the fermentation reaction. As an example, carbon dioxide can be provided as a byproduct of the metabolism of a fermentable carbon source by the microorganism. In some embodiments, desorption with gases can be used simultaneously with the extractant to remove the alcoholic product from the fermentation broth. The alcoholic product can be recovered from the gas phase containing alcoholic product by the use of methods known in the art, such as the use of a cooled water trap to condense the alcoholic product or purify the gas phase with a solvent.

Recombinant microorganisms and biosynthetic routes Without theoretical limitations of any kind, it is believed that the processes described in the present disclosure are useful in conjunction with any microorganism capable of producing fermentation products, which includes alcohol producing microorganisms, particularly, recombinant microorganisms that produce alcohol with titers greater than their own. tolerance levels.

In the matter, microorganisms producing alcohol are known. For example, the fermentative oxidation of methane by methanotrophic bacteria (eg, Methylosinus trichosporium) produces methanol, and the yeast strain CEN.PK113-7D (CBS 8340, the Centraal Buró voor Schimmelculture; van Dijken, et al., Enzyme Microb. Techno 26: 706-714, 2000) produces ethanol. In addition, recombinant microorganisms producing alcohol are known in the art (eg, Ohta, et al., Appl. Environ Microbiol., 57: 893-900, 1991; Underwood, et al., Appl. Environ. Microbiol. : 1071-1081, 2002, Shen and Liao, Metab.Eng.10: 312-320, 2008, Hahnai, et al., Appl. Environ.Microbiol. 73: 7814-7818, 2007, U.S. Pat. No. 5,514,583, U.S. Patent No. 5,712,133, PCT Patent Publication No. O. 1995/028476; Feldmann, et al., Appl. Microbiol. Biotechnol. 38: 354-361, 1992; Zhang, et al., Science 267: 240-243, 1995; US patent application publication UU no. 2007/0031918 Al; US patent UU no. 7,223,575; US patent UU No. 7,741,119; US patent UU no. 7,851,188; US patent application publication UU No..2009 / 0203099 Al; US patent application publication UU no. 2009/0246846 Al; and publication of PCT application no. WO 2010/075241, all incorporated in the present description by reference).

In addition, microorganisms can be modified with the use of recombinant technologies to generate recombinant microorganisms capable of producing alcoholic products such as ethanol and butanol. Microorganisms that can be recombinantly modified to produce an alcoholic product by means of a biosynthetic route include members of the genera Clostridium, Zymomonas, Escherichia, Salmonella, Serratia, Erwinia, Klebsiella, Shigella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klebsiella, Paenibacillus, Arthrobacter, Corynebacterium, Brevibacterium, Schizosaccharomyces, Kluyveromyces, Yarrowia, Pichia, Candida, Hansenula, Issatchenkia or Saccharomyces. In some embodiments, the recombinant microorganisms may be selected from the group consisting of Escherichia coli, Lactobacillus plantarum, Kluyveromyces lactis, Kluyveromyces marxianus and Saccharomyces cerevisiae. In some embodiments, the recombinant microorganism is yeast. In some embodiments, the recombinant microorganism is yeast with a positive "crabtree" effect selected from Saccharomyces, Zygosaccharomyces, Schizosaccharomyces, Dekkera, Torulopsis, Brettanomyces and some Candida species. Yeast species with positive Crabtree effect include, but are not limited to, Saccharomyces cerevisiae, Saccharomyces kluyveri, Schizosaccharomyces po be, Saccharomyces bayanus, Saccharomyces mikitae, Saccharomyces paradoxus, Zygosaccharomyces rouxii and Candida glabrata.

Saccharomyces cerevisiae yeasts are known in the art and can be obtained from various sources including, but not limited to, American Type Culture Collection (Rocville, MD), Centraalbureau voor Schimmelcultures (CBS) Fungal Biodiversity Center, LeSaffre, Gert Strand AB, Ferm Solutions, North American Bioproducts, Martrex and Lallemand. Yeasts of Saccharomyces cerevisiae include, but are not limited to, BY4741, CEN.PK 113-7D, yeast Ethanol Red®, yeast Ferm Pro ™, yeast Bio-Ferm® XR, yeast alcohol Gert Strand Prestige Batch Turbo, yeast Gert Strand Pot Distillers, yeast Gert Strand Distillers Turbo, yeast FerMax ™ Green, yeast FerMax ™ Gold, yeast Thermosacc®, BG-1, PE-2, CAT-1, CBS7959, CBS7960 and CBS7961.

In some embodiments, the microorganism can be immobilized or encapsulated. For example, the microorganism can be immobilized or encapsulated by the use of alginate, calcium alginate or polyacrylamide gels, or by induction of biofilm formation on a variety of support matrices with large surface area, such as diatomite, celite , diatomaceous earth, silica gels, plastics or resins. In some embodiments, ISPR can be used in conjunction with immobilized or encapsulated microorganisms. This combination can improve productivity, such as specific volumetric productivity, metabolic rate, yields of the alcoholic product and tolerance to the alcoholic product. In addition, immobilization and encapsulation can minimize the effects of process conditions, such as shearing, on microorganisms.

The production of butanol with the use of fermentation, as well as microorganisms that produce butanol, is described, for example, in U.S. Pat. UU no. 7,851,188 and the publication of US patent application. UU no. 2007/0092957; 2007/0259410; 2007/0292927; 2008/0182308; 2008/0274525; 2009/0155870; 2009/0305363; and 2009/0305370, the entire contents of which is incorporated herein by reference. In some modalities, the microorganism is developed by genetic engineering so as to contain a biosynthetic pathway. In some modalities, the biosynthetic pathway is a butanol biosynthetic pathway developed by genetic engineering. In some modalities, the biosynthetic route converts pyruvate into a fermentation product. In some modalities, the biosynthetic pathway converts pyruvate, as well as amino acids, into a fermentation product. In some embodiments, at least one, at least two, at least three or at least four polypeptides that catalyze conversions of substrate to product of a route are encoded by heterologous polynucleotides in the microorganism. In some embodiments, all polypeptides that catalyze conversions from substrate to product of a route are encoded by heterologous polynucleotides in the microorganism. In some embodiments, the polypeptide that catalyzes the conversions, from substrate to product, from acetolactate to 2,3-dihydroxyisovalerate and / or the polypeptide that catalyzes the conversion, from substrate to product, from isobutyraldehyde to isobutanol are capable of using nicotinamide adenine dinucleotide reduced (NADH) as a cofactor.

Biosynthetic routes The biosynthetic routes for the production of isobutanol that may be used include those described in US Pat. UU no. 7,851,188, incorporated herein by reference. In one embodiment, the biosynthetic route of isobutanol comprises the following substrate conversions in product: a) pyruvate in acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) acetolactate in 2,3-dihydroxyisovalerate which can be catalyzed, for example, by means of acetohydroxy acid reductoisomerase; c) 2,3-dihydroxyisovalerate in α-ketoisovalerate which can be catalyzed, for example, by means of acetohydroxy acid dehydratase; d) -cetoisovalerate in isobutyraldehyde which can be catalyzed, for example, by means of a branched-chain α-keto acid decarboxylase; and e) isobutyraldehyde in isobutanol which can be catalyzed, for example, by means of a branched chain alcohol dehydrogenase.

In another embodiment, the isobutanol biosynthetic pathway comprises the following substrate to product conversions: a) pyruvate in acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) acetolactate in 2,3-dihydroxyisovalerate which can be catalyzed, for example, by means of keto-acid reductoisomerase; c) 2,3-dihydroxyisovalerate in a-ketoisovalerate, which can be catalyzed, for example, by dihydroxy acid dehydratase; d) α-ketoisovalerate in valine, which can catalyzed, for example, by transaminase or valine dehydrogenase; e) valine in isobutylamine, which can be catalyzed, for example, by valine decarboxylase; f) isobutylamine in isobutyraldehyde, which can be catalyzed, for example, by omega transaminase; Y g) Isobutyraldehyde in isobutanol which can be catalyzed, for example, by means of a branched chain alcohol dehydrogenase.

In another embodiment, the isobutanol biosynthetic pathway comprises the following substrate to product conversions: a) pyruvate in acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) acetolactate in 2,3-dihydroxyisovalerate which can be catalyzed, for example, by means of acetohydroxy acid reductoisomerase; c) 2,3-dihydroxyisovalerate in α-ketoisovalerate which can be catalyzed, for example, by means of acetohydroxy acid dehydratase; d) α-ketoisovalerate in isobutyryl-CoA, which can be catalyzed, for example, by branched chain keto acid dehydrogenase; e) isobutyryl-CoA to isobutyraldehyde, which can be catalyzed, for example, by acylation of the aldehyde dehydrogenase; Y f) isobutyraldehyde in isobutanol which can be catalyzed, for example, by means of a branched chain alcohol dehydrogenase.

The biosynthetic routes for the production of 1-butanol that may be used include those described in the US patent application publication. UU no. 2008/0182308, incorporated herein by reference. In one embodiment, the 1-butanol biosynthetic pathway comprises the following substrate to product conversions: a) acetyl-CoA in acetoacetyl-CoA which can be catalyzed, for example, by means of acetyl-CoA acetyltransferase; b) acetoacetyl-CoA in 3-hydroxybutyryl-CoA which can be catalyzed, for example, by means of 3-hydroxybutyryl-CoA dehydrogenase; c) 3-hydroxybutyryl-CoA in crotonyl-CoA which can be catalyzed, for example, by means of crotonane; d) Crotonyl-CoA in butyryl-CoA which can be catalyzed, for example, by means of butyryl-CoA dehydrogenase; e) Butyryl-CoA in butyraldehyde which can be catalyzed, for example, by means of butyraldehyde dehydrogenase; Y f) butyraldehyde to 1-butanol, which can be catalyzed, for example, by means of butanol dehydrogenase.

The biosynthetic routes for the production of 2-butanol that may be used include those described in the US patent application publication. UU no. 2007/0259410 and the publication of US patent application. UU no. 2009/0155870, incorporated herein by reference. In one embodiment, the 2-butanol biosynthetic pathway comprises the following substrate to product conversions: a) pyruvate in alpha-acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) alpha-acetolactate in acetoin which can be catalyzed, for example, by acetolactate decarboxylase; c) acetoin in 3-amino-2-butanol, which can be catalyzed, for example, by acetonin aminase; d) 3-amino-2-butanol in 3-amino-2-butanol phosphate, which can be catalyzed, for example, by aminobutanol kinase; e) 3-amino-2-butanol phosphate in 2-butanone, which can be catalyzed, for example, by aminobutanol phosphate phosphorylase; Y f) 2-butanone to 2-butanol, which can be catalyzed, for example, by butanol dehydrogenase.

In another modality, the biosynthetic route of 2-butanol the following conversions from substrate to product: a) pyruvate in alpha-acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) alpha-acetolactate in acetoin which can be catalyzed, for example, by acetolactate decarboxylase; c) acetoin in 2,3-butanediol which can be catalyzed, for example, by means of butanediol dehydrogenase; d) 2,3-butanediol in 2-butanone, which can be catalyzed, for example, by dial dehydratase; Y e) 2-butanone in 2-butanol, which can be catalyzed, for example, by butanol dehydrogenase.

The biosynthetic routes for the production of 2-butanone that may be used include those described in the US patent application publication. UU no. 2007/0259410 and in the US patent application publication. UU No..2009 / 0155870, incorporated herein by reference. In one embodiment, the 2-butanone biosynthetic pathway comprises the following substrate conversions into product: a) pyruvate in alpha-acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) alpha-acetolactate in acetoin which can be catalyzed, for example, by acetolactate decarboxylase; c) acetoin in 3-amino-2-butanol, which can be catalyzed, for example, by acetonin aminase; d) 3-amino-2-butanol in 3-amino-2-butanol phosphate, which can be catalyzed, for example, by aminobutanol kinase; Y e) 3-amino-2-butanol phosphate in 2-butanone, which can be catalyzed, for example, by aminobutanol phosphate phosphorylase.

In another embodiment, the biosynthetic route of 2-butanone comprises the following substrate conversions in product: a) pyruvate in alpha-acetolactate which can be catalyzed, for example, by means of acetolactate synthase; b) alpha-acetolactate in acetoin which can be catalyzed, for example, by acetolactate decarboxylase; c) acetoin in 2,3-butanediol which can be catalyzed, for example, by means of butanediol dehydrogenase; Y d) 2,3-butanediol in 2-butanone, which can be catalyzed, for example, by diol dehydratase.

In the present description, the terms "acetohydroxy acid synthase", "Acetolactate synthase" and "acetolactate synthetase" (abbreviated as "ALS") can be used, interchangeably, to refer to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of pyruvate to acetolactate and CO2. Some acetolactate synthases are known as EC 2.2.1.6 (Enzyme Nomenclature 1992, Academic Press, San Diego). These unmodified enzymes are available from many sources including, but not limited to, Bacillus subtilis (GenBank Nos .: CAB15618 (sec.with ident.no.:1), Z99122 (sec.with ident. : 2), amino acid sequence of NCBI (National Center for Biotechnology Information), nucleotide sequence of NCBI, respectively), Klebsiella pneumoniae (GenBank's number: AAA25079 (sec. With ident. No .: 3), M73842 ( sec. with ident. no .: 4)) and Lactococcus lactis (GenBank numbers: AAA25161 (sec. with ident. no .: 5), L16975 (sec. with ident. no .: 6)).

The terms "cetol-acid reductoisomerase" ("KARI"), "acetohydroxy acid isomeroreductase" and "acetohydroxy acid reductoisomerase" can be used interchangeably and refer to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the reaction of (S) -acetolactate to 2,3-dihydroxyisovalerate. Illustrative KARI enzymes can be classified as EC number 1. 1.1.86 (Enzyme Nomenclature 1992, Academic Press, San Diego) and are available from a wide variety of microorganisms, including, but not limited to, Escherichia coli (GenBank Numbers: NP_418222 (sec. .: 7), NC_000913 (sec. With ident. No .: 8), Saccharomyces cerevisiae (GenBank numbers: NP_013459 (sec. With ident. No .: 9), NC_001144 (sec. With no. Ident .: 10)), Methanococcus maripaludis (GenBank numbers: CAF30210 (sec. with ID number: 11), BX957220 (sec. with ident. no .: 12)) and Bacillus subtilis (num. GenBank: CAB14789 (sec. With ident. No .: 13), Z99118 (sec. With ident. No .: 14)). The KARIs include the KARI variants of Anaerostipes caccae "K9G9" and "K9D3" (sec. With ID numbers: 15 and 16, respectively). Cetol-acid reductoisomerase (KARI) enzymes are described in the publications of US patent applications. UU num. 2008/0261230, 2009/0163376 and 2010/0197519 and in the PCT application publication no. WO / 2011/041415, incorporated herein by reference. Examples of KARI described therein are those of Lactococcus lactis, Vibrio cholera, Pseudomonas aeruginosa PAOl and PF5 mutants of Pseudomonas fl ors cens. In some modalities, KARIs use NADH. In some embodiments, KARIs use reduced nicotinamide adenine dinucleotide (NADPH) dinucleotide.

The terms "acetohydroxy acid dehydratase" and "dihydroxy acid dehydratase" ("DHAD") refer to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of 2,3-dihydroxyisovalerate to a-ketoisovalerate. Some acetohydroxy acid dehydratases are known as EC 4.2.1.9. These enzymes are available from a wide variety of microorganisms including, but not limited to, Escherichia coli (GenBank Nos: YP_026248 (sec. With ident. No .: 17), NC000913 (sec. With ident. .: 18)), Saccharomyces cerevisiae (GenBank numbers: NP_012550 (sec. With ident. No .: 19), NC 001142 (sec. With ident. No .: 20), M. maripaludis (no. GenBank: CAF29874 (sec. With ident. No .: 21), BX957219 (sec. With ident. No .: 22), B. Subtilis (GenBank numbers: CAB14105 (sec. With ident. : 23), Z99115 (sec. With ID No.: 24)), L. lactis and N. crassa US Patent Application Publication No. 2010/0081154 and the US Patent No. 7,851,188, which are incorporated herein by reference, disclose dihydroxy acid dehydratases (DHAD), which include a DHAD of Streptococcus mutans.

The terms "branched-chain α-ketoacid decarboxylase", "α-ketoacid decarboxylase", "α-ketoisovalerate decarboxylase" or "2-ketoisovalerate decarboxylase" ("KIVD") refer to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of α-ketoisovalerate to isobutyraldehyde and CO2.

Illustrative branched-chain α-ketoacid decarboxylases are known as EC 4.1.1.72 and are available from many sources, including, but not limited to, Lactococcus lactis (GenBank Nos .: AAS49166 (Seq. Ident .: 25), AY548760 (sec. with ID: 26); CAG34226 (sec. with ID: 27), AJ746364 (sec. with ID: 28), Salmonella typhimurium (GenBank nos .: NP_461346 (sec. with ID: 29), NC_003197 (sec.with ident.no.:30)), Clostridium acetobutylicum (GenBank nos .: NP_149189 (sec. Ident .: 31), NC_001988 (sec. with ident. no .: 32), M. caseolyticus (sec. with ident. no .: 33) and L. grayi (sec. with ident. : 3. 4).

The term "branched chain alcohol dehydrogenase" ("ADH") refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of isobutyraldehyde to isobutanol. Some branched-chain alcohol dehydrogenases are known as EC 1.1.1.265, but can be further classified under other numbers of alcohol dehydrogenases (specifically, EC 1.1.1.1 or 1.1.1.2). Alcohol dehydrogenases may be NADPH dependent or NADH dependent. These enzymes are available from many sources including, but not limited to, Saccharomyces cerevisiae (GenBank #: NP_010656 (sec. with no. Ident .: 35), NC_001136 (sec. with ID number: 36), NP_014051 (sec. with ID number: 37), NC_001145 (sec. with ID number: 38)), Escherichia coli (GenBank numbers: NP_417484 (sec. With ident. No .: 39), NC_000913 (sec. With ident. No .: 40)), C. acetobutylicum (GenBank numbers: NP_349892 (sec. with ID number: 41), NC_003030 (ident. number with ID: 42); NP_349891 (with ID: 43), NC_003030 (with ID: 44 )). The publication of US patent application UU no. 2009/0269823 describes SadB, an alcohol dehydrogenase (ADH) of Achromobacter xylosoxidans. The alcohol dehydrogenases also include ADH from horse liver and ADH from Beijerinkia indica (as described in U.S. Patent Application Publication No. 2011/0269199, which is incorporated herein by reference).

The term "butanol dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of isobutyraldehyde to isobutanol or the conversion of 2-butanone and 2-butanol. The butanol dehydrogenases are a subset of a large family of alcohol dehydrogenases. The butanol dehydrogenase can be dependent on NAD or NADP. The NAD-dependent enzymes are known as EC 1.1.1.1 and are available, for example, from Rhodococcus ruber (GenBank Nos .: CAD36475, AJ491307). NADP-dependent enzymes are known as EC 1. 1.1.2 and are available, for example, from Pyrococcus furiosus (GenBank Num: AAC25556, AF013169). In addition, a butanol dehydrogenase is available from Escherichia coli (GenBank Nos .: NP 417484, NC_000913) and a cyclohexanol dehydrogenase is available from Acinetobacter sp. (GenBank numbers: AAG10026, AF282240). The term "butanol dehydrogenase" also refers to an enzyme that catalyzes the conversion of buliraldehyde to 1-butanol, with the use of NADH or NADPH as a cofactor. Butanol dehydrogenases are available, for example, from C. acetobutilicum (GenBank Nos: NP_149325, NC_001988, this enzyme possesses aldehyde and alcohol dehydrogenase activity); NP_349891, NC_003030 and NP_349892, NC_003030) and Escherichia coli (GenBank Num: NP_417-484, NC_000913).

The term "branched chain ketoacid dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of α-ketoisovalerate to isobutyryl-CoA (isobutyryl-coenzyme A), typically, with the use of NAD + (nicotinamide) adenine dinucleotide) as an electron acceptor. Some branched-chain keto-acid dehydrogenases are known as EC 1.2.4.4. Such branched chain keto acid dehydrogenases are composed of four subunits and the sequences of all subunits are available from a wide variety of microorganisms including, but not limited to, limit to, Bacillus subtilis (GenBank's number: CAB14336 (sec. with ID number: 45), Z99116 (sec. with ID: 46); CAB14335 (sec. with ident. : 47), Z99116 (sec. With ID number: 48), CAB14334 (sec. With ID number: 49), Z99116 (sec. With ID number: 50); and CAB14337 (sec. with ID: 51), Z99116 (sec. with ID No.: 52)) and Pseudomonas putida (GenBank's number: AAA65614 (sec. with ID: 53), M57613 ( sec with ID number: 54); AAA65615 (sec. with ID: 55), M57613 (sec. with ID: 56); AAA65617 (sec. with ident. : 57), M57613 (sec. With ID: 58), and AAA65618 (sec. With ID: 59), M57613 (sec. With ID: 60)).

The term "acylating aldehyde dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of isobutyryl-CoA to isobutyraldehyde, typically, with the use of NADH or NADPH as the electron donor. Some acylating aldehyde dehydrogenases are known as EC 1.2.1.10 and 1.2.1.57. Such enzymes are available from multiple sources including, but not limited to, Clostridium beijerinckii (GenBank Nos .: AAD31841 (sec. With ident. No .: 61), AF157306 (sec. With ident. )), Clostridium acetobutylicum (GenBank numbers: NP_149325 (sec. With ident. No .: 63), NC_001988 (sec. With ident. No .: 64), PN 149199 (sec. With ident. : 65), NC 001988 (sec. With identity number: 66)), Pseudomonas putida (GenBank nos .: AAA89106 (sec. With ident. No .: 67), U13232 (sec. With ident. No .: 68)) and Thermus thermophilus (GenBank nos .: YP_145486 (sec. ID number: 69), NC_006461 (sec. with ID number: 70)).

The term "transaminase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of α-ketoisovalerate to L-valine, with the use of alanine or glutamate as amine donors. Some transaminases are known with the numbers EC 2.6.1.42 and 2.6.1.66. Such enzymes are available from numerous sources. Examples of sources for alanine-dependent enzymes include, but are not limited to, Escherichia coli (GenBank Nos: YP_026231 (sec. With ident. No .: 71), NC_000913 (sec. With ident. No .: 72)) and Bacillus licheniformis (GenBank nos .: YP_093743 (sec. With ident. No .: 73), NC_006322 (sec. With ident. No .: 74)). Examples of sources for glutamate-dependent enzymes include, but are not limited to, Escherichia coli (GenBank Nos: YP_026247 (sec. With ident. No .: 75), NC_000913 (sec. With ident. No .: 76)), Saccharomyces cerevisiae (GenBank numbers: NP_012682 (sec. With ident. No .: 77), NC_001142 (sec. With ident. No .: 78)) and Methanobacterium thermoautotrophicum (GenBank numbers: NP_276546 (sec. with ID number: 79), NC_000916 (sec. with ID number: 80)).

The term "valine dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of α-ketoisovalerate to L-valine, typically, with the use of NAD (P) H as an electron donor and ammonia as an amine donor. Some valine dehydrogenases are known with EC numbers 1.4.1.8 and 1.4.1.9 and such enzymes are available from numerous sources including, but not limited to, Streptomyces coelicolor (GenBank numbers: NP_628270 (sec. With ident. .: 81), NC_003888 (sec. With ID: 82)) and Bacillus subtilis (GenBank numbers: CAB14339 (sec. With ident. No .: 83), Z99116 (sec. With ident. No .: 84)).

The term "valine decarboxylase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of L-valine to isobutylamine and CO2. Some valine decarboxylases are known as EC 4.1.1.14. Such enzymes are found in Streptomyces, such as, for example, Streptomyces viridif ciens (GenBank nos .: AA 10242 (sec.with ident.no .: 85), AY116644 (sec. With ident. No .: 86)).

The term "omega transaminase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of isobutylamine to isobutyraldehyde, with the use of a suitable amino acid as an amine donor. Illustrative omega transaminases are known as EC 2.6.1.18 and are available from many sources, including, but not limited to, Alcaligenes denitrificans (AAP92672 (sec. with ident. no .: 87), AY330220 (sec.with ident.ind .: 88)), Ralstonia eutropha (num. GenBank: YP_294474 (sec. With ident. No .: 89), NC_007347 (sec. With ident. No .: 90)), Shewanella oneidensis (GenBank numbers: NP_719046 (sec. With ident. No .: 91), NC_004347 (sec. With ID: 92)) and Pseudomonas putida (GenBank nos .: AAN66223 (sec. With ID: 93), AE016776 (sec. With ident. : 94)).

The term "acetyl-CoA acetyltransferase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of two molecules of acetyl-CoA into acetoacetyl-CoA and coenzyme A (CoA). Examples of acetyl-CoA acetyltransferases are acetyl-CoA acetyltransferases with substrate preferences (reaction in the forward direction) for a short-chain acyl-CoA and acetyl-CoA and are classified with the number EC2.3.1.9 [Enzyme Nomenclature 1992 , Academic Press, San Diego]; However, enzymes with a wider substrate range (E.C.2.3.1.16) will also be functional. Acetyl-CoA acetyltransferases are available from many sources, for example, Escherichia coli (GenBank No.s: NP_416728, NC_000913, NCBI amino acid sequence, NCBI nucleotide sequence), Clostridium acetobutylicum (GenBank No.: NP 349476.1, NC 003030; NP 149242, NC 001988, Bacillus subtilis (GenBank No.: PN 390297, NC_000964) and Saccharomyces cerevisiae (GenBank numbers: NP_015297, NC_001148).

The term "3-hydroxybutyryl-CoA dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of acetoacetyl-CoA to 3-hydroxybutyryl-CoA. Examples of 3-hydroxybutyryl-CoA dehydrogenases may be NADH-dependent, with a substrate preference for (S) -3-hydroxybutyryl-CoA or (R) -3-hydroxybutyryl-CoA. The examples can be classified as E.C. 1.1.1.35 and E.C.1.1.1.30, respectively. In addition, the 3-hydroxybutyryl-CoA dehydrogenases may be NADPH-dependent, with a substrate preference for (S) -3-hydroxybutyryl-CoA or (R) -3-hydroxybutyryl-CoA and are classified as EC1.1.1.157 and EC 1.1.1.36, respectively. 3-Hydroxybutyryl-CoA dehydrogenases are available from many sources, for example, Clostridium acetobutylicum (GenBank numbers: NP_349314, NC_003030), Bacillus subtilis (GenBank numbers: AAB09614, U29084), Ralstonia eutropha (GenBank numbers: YP_294481, NC_007347) and Alcaligenes eutrophus (GenBank nos .: AAA21973, J04987).

The term "crotonane" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of 3-hydroxybutyryl-CoA to crotonyl-CoA and H2O. Illustrative crotonates may show a substrate preference for (S) -3-hydroxybutyryl-CoA or (R) -3- hydroxybutyryl-CoA and can be classified as E.C.4.2.1.17 and E.C. 4.2.1.55, respectively. Crotonades are available from many sources, for example, Escherichia coli (GenBank No.: NP_415911, NC_000913), Clostridium acetobutylicum (GenBank No.: NP_349318, NC_003030), Bacillus subtilis (GenBank No.: CAB13705, Z99113) and Aeromonas caviae (GenBank numbers: BAA21816, D88825).

The term "butyryl-CoA dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of crotonyl-CoA to butyryl-CoA. Examples of butyryl-CoA dehydrogenases can be NADH-dependent, NADPH-dependent or flavin-dependent and can be classified as E.C. 1.3.1.44, E.C. 1.3.1.38 and E.C. 1.3.99.2, respectively. Butyryl-CoA dehydrogenases are available from many sources, eg, Clostridium acetobutylicum (GenBank Num: NP_347102, NC_003030), Euglena gracilis (GenBank Num: Q5EU90, AY741582), Streptomyces collinus (GenBank Num: AAA92890, U37135) and Streptomyces coelicolor (GenBank numbers: CAA22721, AL939127).

The term "butyraldehyde dehydrogenase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of butyryl-CoA to butyraldehyde, with the use of NADH or NADPH as a cofactor. The buliraldehyde dehydrogenases with a preference for NADH they are known as E.C. 1.2.1.57 and are available, for example, from Clostridium beijerinckii (GenBank Num: AAD31841, AF157306) and Clostridium acetobutylicum (GenBank Num: NP.sub 149325, NC.sub .-- 001988).

The term "isobutyryl-CoA mutase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of butyryl-CoA to isobutyryl-CoA. This enzyme uses coenzyme B12 as a cofactor. Some isobutyryl-CoA mutants are known as EC 5.4.99.13. These enzymes are found in various Streptomyces, including, but not limited to, Streptomyces cinnamonensis (GenBank Nos .: AAC08713 (sec. With ident. No .: 95), U67612 (sec. With ident. No .: 96), CAB59633 (sec. With ID No. 97), AJ246005 (sec. With ID No. 98), Streptomyces coelicolor (GenBank Nos .: CAB70645 (sec. With ident. : 99), AL939123 (sec. With ID: 100); CAB92663 (sec. With ID: 101), AL939121 (sec. With ID: 102)) and Streptomyces avermitilis ( GenBank numbers: NP_824008 (sec. with ID number: 103), NC_003155 (sec. with ID number: 104); NP_824637 (sec. with ID number: 105), NC_003155 (sec. with identification number: 106)).

The term "acetolactate decarboxylase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of alpha-acetolactate to acetoin. Some acetolactate decarboxylases are known by the number EC 4.1.1.5 and are available, for example, from Bacillus subtilis (GenBank numbers: AAA22223, L04470), Klebsiella terrigena (GenBank numbers: AAA25054, L04507) and Klebsiella pneumoniae (GenBank numbers: AAU43774, AY722056) .

The terms "acetoin aminase" or "acetoin transaminase" refer to a polypeptide (or polypeptides) with enzymatic activity that catalyzes the conversion of acetoin to 3-amino-2-butanol. Acetoin aminase can use the cofactor pyridoxal 5'-phosphate or NADH or NADPH. The resulting product may have stereochemistry (R) or (S) at position 3. The pyridoxal phosphate-dependent enzyme may use an amino acid, such as alanine or glutamate, as an amino donor. The NADH and NADPH-dependent enzymes can use ammonia as the second substrate. A suitable example of an NADH-dependent acetoin aminase, also known as amino alcohol dehydrogenase, is described in Ito et al. (U.S. Patent No. 6,432,688). An example of pyridoxal-dependent acetoin aminase is the amino: pyruvate aminotransferase (also called amino: pyruvate transaminase), described by Shin and Kim (J. Org.Chem.67: 2848-2853, 2002).

The term "acetoin kinase aminase" refers to a polypeptide (or polypeptides) with an enzymatic activity that catalyzes the conversion of acetoin to phosphoacetoin. Acetoin kinase can use ATP (adenosine triphosphate) or phosphoenolpyruvate as the phosphate donor in the reaction.

Enzymes that catalyze the analogous reaction in the similar substrate of dihydroxyacetone, for example, include the enzymes known as EC 2.7.1.29 (Garcia-Alles, et al., Biochemistry 43: 13037-13046, 2004).

The term "acetoin phosphate aminase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of phosphoacetoin to 3-amino-2-butanol O-phosphate. Acetoin phosphate aminase can use the cofactor pyridoxal 5'-phosphate, NADH or NADPH. The resulting product may have stereochemistry (R) or (S) at position 3. The pyridoxal phosphate-dependent enzyme may use an amino acid, such as alanine or glutamate. The NADH and NADPH-dependent enzymes can use ammonia as the second substrate. Although there are no reports of enzymes that catalyze this reaction in phosphoacetoin, the use of a pyridoxal phosphate-dependent enzyme is proposed to perform the analogous reaction in the similar substrate of serinol phosphate (Yasuta, et al., Appl. Environ. Microbiano .67: 4999-5009, 2001).

The term "aminobutanol phosphate phospholiase", also known as "amino alcohol O-phosphate lyase", refers to a polypeptide (or polypeptides) with an enzymatic activity that catalyzes the conversion of 3-amino-2-butanol O-phosphate in 2-butanone. The amino butanol phosphate phospho-lyase can use the cofactor pyridoxal'-5-phosphate. There are reports about enzymes that catalyze the analogous reaction in the similar substrate 1- amino-2-propanol phosphate (Jones, et al., Biochem J.134: 167-182, 1973). The publication of US patent application UU num.2007 / 0259410 describes an aminobutanol phosphate phospholipase from the Erwinia carotovora organism.

The term "aminobutanol kinase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the conversion of 3-amino-2-butanol to 3-amino-2-butanol 0-phosphate. The amino butanol kinase can use ATP as a phosphate donor. While there are no reports of enzymes that catalyze this reaction in 3-amino-2-butanol, there are reports of enzymes that catalyze the analogous reaction in the similar substrates ethanolamine and 1-amino-2-propanol (Jones, et al., supra). The publication of US patent application UU no. 2009/0155870 describes, in Example 14, an amino alcohol kinase from Erwinia carotovora subsp. Atroseptica.

The term "butanediol dehydrogenase", also known as "acetoin reductase", refers to a polypeptide (or polypeptides) with enzymatic activity that catalyzes the conversion of acetoin to 2,3-butanediol. Butanodial dehydrogenases are a subset of the large family of alcohol dehydrogenases. The butanediol dehydrogenase enzymes may have specificity for the production of stereochemistry (R) or (S) in the alcohol product. The butanediol dehydrogenases specific for (S) are known as EC 1.1.1.76 and are available, for example, Klebsiella pneumoniae (GenBank Num: BBA13085, D86412). The specific butanediol dehydrogenases of (R) are known by the number EC 1.1.1.4 and are available, for example, from Bacillus cereus (GenBank No.s. NP 830481, NC_004722; AAP07682, AE017000) and Lactococcus lactis (GenBank numbers: AAK04995, AE006323).

The term "butanediol dehydratase", further known as "dial dehydratase" or "propanediol dehydratase" refers to a polypeptide (or polypeptides) with enzymatic activity that catalyzes the conversion of 2,3-butanediol to 2-butanone. Butanediol dehydratase may use the co-factor adenosyl cobalamin (also known as coenzyme Bw or vitamin B12, although vitamin B12 may also refer to other forms of cobalamin that are not coenzyme B12). Enzymes dependent on adenosyl cobalamin are known as EC 4.2.1.28 and are available, for example, from Klebsiella oxytoca (GenBank's Num: AA08099 (alpha subunit), D45071; BAA08100 (beta subunit), D45071; and BBA08101 (gamma subunit), D45071 (all three subunits are required for activity) and Klebsiella pneumonia (GenBank numbers: AAC98384 (alpha subunit), AF102064; GenBank numbers: AAC98385 (beta subunit), AF102064, nos. from GenBank: AAC98386 (gamma subunit), AF102064). Other suitable dehydratase dialings include, but are not limited to, B12-dependent dial dehydratases available from Salmonella typhimurium (GenBank numbers: AAB84102 (large subunit), AF026270; num. from GenBank: AAB84103 (medium subunit), AF026270; num. from GenBank: AAB84104 (small subunit), AF026270); and Lactobacillus collinoides (GenBank No.: CAC82541 (large subunit), AJ297723; GenBank No.: CAC82542 (medium subunit); AJ297723; GenBank No.: CAD01091 (small subunit), AJ297723); and Lactobacillus brevis enzymes (particularly, strains CNRZ 734 and CNRZ 735, Speranza, et al., J. Agrie, Food Chem. 45: 3476-3480, 1997) and nucleotide sequences encoding the corresponding enzymes. Methods for isolation of the dial dehydratase gene are well known in the art (for example, U.S. Patent No. 5,686,276).

The term "pyruvate decarboxylase" refers to a polypeptide (or polypeptides) having enzymatic activity that catalyzes the decarboxylation of pyruvic acid in acetaldehyde and carbon dioxide. Pyruvate dehydrogenases are known as EC 4.1.1.1. These enzymes are found in various yeasts that include Saccharomyces cerevisiae (GenBank nos .: CAA97575 (sec. With ident. No .: 107), CAA97705 (sec. With ident. No .: 109), CAA97091 (sec. Ident. no .: 111)).

It will be appreciated that microorganisms comprising an isobutanol biosynthetic pathway, as provided in the present invention, may further comprise one or more additional modifications. The publication of US patent application UU No..2009 / 0305363 (incorporated by reference) describes the increased conversion of pyruvate to acetolactate by the genetic modification of yeast for the expression of an acetolactate synthase located in the cytosol and the substantial elimination of pyruvate decarboxylase activity. In some embodiments, the microorganisms may comprise modifications to reduce the activity and / or interruption of the glycerol-3-phosphate dehydrogenase in at least one gene encoding a polypeptide having pyruvate decarboxylase activity or an interruption in at least one gene encoding a regulatory element that controls the expression of the pyruvate decarboxylase gene, as described in the US patent application publication. UU no. 2009/0305363 (incorporated herein by reference) and / or modifications that provide increased carbon flux through an Entner-Doudoroff route or reduction of the balance of equivalents, as described in the US patent application publication. UU no. 2010/0120105 (incorporated herein by reference). Other modifications include the integration of at least one polynucleotide encoding a polypeptide that catalyzes a step in a biosynthetic pathway using pyruvate. Other modifications include at least one deletion, mutation and / or substitution in an endogenous polynucleotide encoding a polypeptide with acetolactate reductase activity. In some embodiments, the polypeptide having acetolactate reductase activity is YMR226C (sec. With identity numbers: 127, 128) of Saccharomyces cerevisiae or a homolog thereof. Additional modifications include a deletion, mutation and / or substitution in an endogenous polynucleotide that encodes a polypeptide with aldehyde dehydrogenase and / or aldehyde oxidase activity. In some embodiments, the polypeptide having aldehyde dehydrogenase activity is ALD6 of Saccharomyces cerevisiae or a homologue thereof. A genetic modification that has the effect of reducing glucose suppression, wherein the yeast production host cell is pdc- is described in the US patent application publication. UU no. 2011/0124060 incorporated in the present description as a reference. In some embodiments, the pyruvate decarboxylase that is eliminated or downregulated is selected from the group consisting of: PDC1, PDC5, PDC6, and combinations thereof. In some embodiments, pyruvate decarboxylase is selected from the enzymes of Table 1. In some embodiments, microorganisms may contain a deletion or down-regulation of a polynucleotide that encodes a polypeptide that catalyzes the conversion of glyceraldehyde-3-phosphate to 1,3 glycerate, bisphosphate. In some embodiments, the enzyme that catalyzes this reaction is glyceraldehyde-3-phosphate dehydrogenase. Table 1. Sec. With ident numbers. of coding regions and proteins of PDC target gene In some embodiments, any specific nucleic acid molecule or polypeptide can be at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence or polypeptide sequence described in the present description. The phrase "percent identity", as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relationship between the polypeptide or polynucleotide sequences, as the case may be, as determined by the match between the strings of the sequences. Identity and similarity can be easily calculated by known methods including, but not limited to, those described in: Computational Molecular Biology (Lesk, A.M., Ed.) Oxford University: NY (1988); Biocomputing: Informatics and Genome Projects (Smith, D.W., Ed.) Academic: NY (1993); Computer Analysis of Seguence Data, Part I (Griffin, A.M., and Griffin, H.G., Eds.) Humania: NJ (1994); Seguence Analysis in Molecular Biology (von Heinje, G., Ed.) Academic (1987); and Seguence Analysis Primer (Gribskov, M. and Devereux, J, Eds.) Stockton: NY (1991).

The methods for determining identity are designed to give the best match between the sequences under test. The methods for determining identity and similarity are codified in publicly available software. Calculations of sequence alignments and identity percentages can be made with the MegAlign ™ program of the set of bioinformatics programs LASERGENE (DNASTAR Inc., Madison, I). The multiple alignment of the sequences is done with the Clustal alignment method, which covers different varieties of the algorithm that include the Clustal V alignment method that corresponds to the alignment method identified as Clustal V (described by Higgins and Sharp, CABIOS.5: 151-153, 1989, Higgins, et al., Comput. Appl. Biosci. 8: 189-191, 1992) and included in the MegAlign ™ program of the bioinformatics program LASERGENE (DNASTAR Inc.). For multiple alignments, the default values correspond to PENALTY OF INTERRUPTION = 10 and PENALIZATION BY LENGTH OF INTERRUPTION = 10. The default parameters for alignments in pairs and calculation of the percent identity of protein sequences with the use of the Clustal method are KTUPLE = 1, PENALTY OF INTERRUPTION 3, WINDOW = 5 and DIAGONALS SAVED = 5. In the case of nucleic acids, these parameters are KTUPLE = 2, PENALTY OF INTERRUPTION = 5, WINDOW = 4 and DIAGONALS SAVED = 4. After the alignment of the sequences with the use of the Clustal V program, it is possible to obtain a percentage of identity from the table of distances of the sequence in the same program. In addition, the Clustal W alignment method is available and corresponds to the alignment method identified as Clustal W (described by Higgins and Sharp, CABIOS. 5: 151-153, 1989; Higgins, et al., Comput. Appl. Biosci. 8: 189-191, 1992) and included in the program MegAlign ™ v6.1 of the bioinformatics program LASERGENE (DNASTAR Inc.). Default parameters for multiple alignment (GAP PENALTY = 10, PENALTY BY LENGTH OF GAP = 0.2, delay of divergent sequences (%) = 30, DNA transition weight = 0.5, weight matrix for proteins = Gonnet series, matrix of weights for the DNA = IUB). After aligning the sequences with the use of the Clustal W program, it is possible to obtain an identity percentage from the sequence distance table in the same program.

The standard techniques of recombinant DNA and molecular cloning are well known in the art and are described in Sambrook, et al. (Sambrook, J., Fritsch, EF and Maniatis, T. (Molecular Cloning: A Laboratory Manual, Coid Spring Harbor Laboratory Press, Coid Spring Harbor, 1989, which in the present description is referred to as Maniatis) and by Ausubel, et al. (Ausubel, et al., Current Protocols in Molecular Biology, published by Greene Publishing Assoc. And Wilcy-Interscience, 1987.) Some examples of methods for producing microorganisms comprising a butanol biosynthetic pathway are described, for example, in the U.S. Patent No. 7,851,188 and U.S. Patent Application Publication Nos. 2007/0092957, 2007/0259410, 2007/0292927; 2008/0182308; 2008/0274525; 2009/0155870; 2009/0305363; and 2009/0305370, the entire contents of which is incorporated herein by reference.

Furthermore, while various embodiments of the present invention have been described earlier in the present description, it should be understood that they have been presented only as examples, without being limiting. It will be apparent to persons with experience in the pertinent technique that various changes in the form and details of this can be made without departing from the spirit and scope of the invention. Therefore, the coverage and scope of the present invention should not be limited to any of the illustrative embodiments described above, but should be defined only in accordance with the claims and their equivalents.

All publications, patents and patent applications mentioned in this description are indicative of the level of knowledge of the person skilled in the art to which this invention pertains and are incorporated in the present description as a reference for all purposes as if specifically and individually indicated that each publication, patent or individual patent application is incorporated as a reference.

EXAMPLES The following non-limiting examples will further illustrate the invention. It should be understood that, although maize is used as a raw material in the following examples, other sources of biomass, such as cane, for raw materials are used without departing from the present invention. In addition, while the following examples relate to ethanol and butanol, other alcohols or fermentation products can be produced without departing from the present invention.

The meaning of the abbreviations is as follows: "atm" means atmosphere, "ccm" means cubic centimeter (s) per minute, "g / 1" means gram (s) per liter, "g" means grams (s) ), "Gpl" means gram (s) per liter, "gpm" means gallon (s) per minute, "h" means time (s), "HPLC" refers to high performance liquid chromatography, "kg" means kilogram (s), "1" means liter (s), "min" means minute (s), "mi" means milliliter (s), "ppm" means parts per million, "psig" means pounds (s) per square inch pressure gauge and "% by weight" means percent by weight.

Example 1 Process for the production and recovery of butanol produced by fermentation The processes described above can be demonstrated by computer modeling, such as Aspen modeling (see, e.g., U.S. Patent No. 7,666,282). For example, the commercial modeling application Aspen Plus® (Aspen Technology, Inc., Burlington, MA) can be used in conjunction with databases of physical properties, such as DIPPR, available from American Institute of Chemical Engineers, Inc. (New York, NY) to develop an Aspen model for an integrated process of butanol fermentation, purification, and water management. This modeling of the process can perform any fundamental engineering calculation, for example, mass and energy balance, vapor / liquid equilibrium and reaction rate calculations. To generate an Aspen model, the data that must be entered may include, for example, experimental data, water content and composition of the raw material, firing temperature of the pasta and instant evaporation, saccharification conditions (for example, enzyme feeding , starch conversion, temperature, pressure), fermentation conditions (eg, feeding microorganisms, glucose conversion, temperature, pressure), degassing conditions, solvent columns, pre-evaporation columns, condensers, evaporators, centrifuges, etc.

An Aspen model with rigorous material and energy balance was developed, in which 53400 kg / h of corn was crushed and fermented to produce isobutanol and in which most of the isobutanol was extracted during the fermentation and distilled. This model included an approximation of sequential batch fermentations as continuous processes. An example of this fermentation, extraction and distillation process is illustrated in Figure 14.

The 601 liquefied corn templa that was clarified to comprising 1.5% by weight of suspended solids was pumped at 1701 kN / h (170.7 tons / h) and 85 ° C by a heat exchanger and a water cooler, and fermentation was introduced at 600 to 32 ° C. Steam stream 602 was vented at 171 kN / h (17.2 tons / h) at atmospheric pressure from fermentation 600 to a scrubber, with an average continuous molar composition of 95.8% carbon dioxide, 3.4% water and 0.8% of isobutanol. An average beer stream 603 comprising 12.6 gpl of isobutanol was continuously discharged from fermentation 600 and preheated in a heat exchanger by temper 601, before being distilled to recover isobutanol.

Stream 604 with 38611 kN / h (3875 tons / h) of combined average flow was removed from fermentation 600 at an average isobutanol concentration of 11.1 gpl and an average temperature of 32 ° C, and circulated through the 610 extractor for the partial removal of isobutanol. The aqueous salient broth 605 containing 7.9 gpl of isobutanol was cooled by heat exchange with water from the chiller tower (CTW) to 30 ° C before re-entering 600 fermentation. comprising diisopropylbenzene entered the extractant 610 and came out as stream 606 comprising 30.1 gpl of isobutanol. The 610 extractor effectively provided five theoretical stages of liquid-liquid equilibrium to bring the broth into contact of fermentation with the solvent. Current 606 passed at 3388 kN / h (340 tons / h) through a heat exchanger and entered the middle part of twelve theoretical stages of distillation column 620. A steam boiler operated at 0.06 MPa (0.6 atm) and 183 ° C with the use of steam at 1.03 MPa (150 psig) to produce a stream of solvent 607 comprising diisopropylbenzene and essentially devoid of isobutanol which exchanged heat with solvent stream 606 by means of a heat exchanger and cooled still more by cooling water CTW before re-entering the 610 extractor. The overhead steam from the distillation column 620 was cooled with CTW and condensed at 630 to form 230 kN / h (23.1 tons / h) reflux, 1.9 kN / h (0.2 tons / h) of an exhaust gas exhaust vapor 608 and 132 kN / h (13.2 tons / h) of product distillate 609 comprising 99.2% isobutanol, 0.6% water and 0.2% diisopropylbenzene.

Example 2 Process for the recovery of ethanol through the use of an extractant column To process the fermentation broth produced during ethanol fermentation, a Karr® 2.5 cm (1") diameter extraction column was used (Koch Modular Process Systems, Paramus, NJ) .The column contains a series of plates arranged at length of it, which are connected to an axis central. The shaft is connected to a traction unit that can move the perforated plates (perforations 0.64 cm (1/4") in diameter) up and down in an alternate motion.The frequency of the movement was a variable during the test, but both the stroke length of the oscillation (1.9 cm (0.75")) and the separation of the trays (5.1 cm (2")) remained fixed.The column used had a plate pile height of 3000 mm.

An aqueous feed consisting of fermentation broth was supplied from the top of the column, while a corn oil fatty acid (COFA) feed was supplied as an extractant through the bottom of the column. The two feeds flowed down the column in countercurrent to each other and were collected as product at the opposite ends of the column.

The fermentation broth was obtained through the use of a fermentation protocol for the production of ethanol from liquefied and saccharified corn temples from which, in some cases, some of the solids had been removed by means of centrifugation. In some cases, the extraction tests were carried out over several days, so that a portion of the tests were carried out while the CO2 release was at or near its maximum, while another portion was made when the release of CO2 was carried out. Gas had stopped effectively. The COFAs used in this work were of degree Distilled, from Emery Oleochemicals (Cincinnati, OH).

Some experiments were performed with COFA as continuous phase of the column, while others were performed with continuous aqueous phase. The experiments were also carried out with or without internal components installed. Two types of internal components were evaluated: stainless steel and polytetrafluoroethylene (PTFE). A range of flow rates was examined to determine the flow regimes in which the column could function without flooding.

Impact of dynamic feeding from fermentation During the course of the tests, it was determined that, in some cases, the performance of the column varied with the progress of the fermentation. In the initial stages of fermentation, the fermentation broth comprising the feed has a high sugar content; in intermediate stages, a considerable amount of CO2 (which can affect fluid flows) is released from the fermentation broth, whereas in advanced stages the concentration of ethanol in the fermentation broth is high. This temporary variation in the feeding was reflected in the variations in the capacity of the extraction column.

Under the conditions using PTFE plates and COFA continuous phase (without agitation), a difference in performance was observed when using fermentation broth collected at the time when the fermentation was close to the period of maximum gas evolution ("intermediate broth") and towards the end of the fermentation ("final broth"). With the use of the final broth, a liquid throughput speed of 570 lpm / m2 (14 gpm / ft2) (Sample 3E) was achieved without flooding the column. The maximum yield that could be achieved for the intermediate broth before the flood was less than 367 bpm / m2 (9 gpm / ft2) (Sample 4D), with notable differences in the size and appearance of the aqueous droplets. The droplet size of the aqueous phase was higher (with the formation of globules) in the final broth compared to the intermediate broth.

Continuous phase The maximum performance of the column was also affected by the nature of the continuous phase. In the conditions at the end of the fermentation, in an experiment with continuous aqueous phase and internal components of stainless steel (stainless steel), a total liquid capacity of almost 570 lpm / m2 (14 gpm / ft2) was achieved (Sample 2B ). In the case of continuous organic phase and internal components of PTFE, the velocity was less than 367 bpm / m2 (9 gpm / ft2) (Sample 4D). The results are shown in Table 2. The abbreviation AQ refers to the aqueous phase and the abbreviation ORG refers to the organic phase. With reference to Table 2, the Phase was continuous, Sample refers to experimental conditions, Internal components refers to the material of the internal components, Nom. AQ refers to the nominal aqueous flow, Nom. ORG refers to the nominal organic flow, Total flow (ccm) refers to the total flow of the aqueous and organic feeds, and Total flow (gpm / ft2) refers to the total flow per unit area in cross section.

Table 2 When the column operated without internal components by using feed composed of fermentation broth near the end of the fermentation, the choice of the continuous phase affected the capacity of the column. With the continuous aqueous phase, it was possible to operate at approximately 1019 lpm / m2 (25 gpm / ft2) (Samples 2G and 2H). With the continuous phase of COFA, however, it produced flood problems at 733 bpm / m2 (18 gp / ft2) (Sample 21). The results are shown in Table 3.

Table 3 Effect of fermentation conditions on the extraction capacity of the column The nature of the fermentation broth is not static, but changes with the progress of the fermentation process. In fermentation, the concentration of carbohydrates decreases as these are metabolized by microorganisms. This change in composition in the fermentation broth alters physical parameters, such as viscosity and surface tension of the fermentation broth, which have an effect on the extraction process. In addition to the changes in concentration, in the intermediate stages a considerable amount of CO2 is released; and this CO2 affects the flow of aqueous and organic liquids through the column.

To process the fermentation broth from an ethanol fermentation, a Karr® glass extraction column was used., 2.5 cm (1") diameter (Koch Modular Process Systems, Paramus, NJ), equipped with internal PTFE components, processing was performed at various time points during the course of the fermentation, the organic extractant (COFA) was the continuous phase of the column and the fermentation broth passed through the column in the form of droplets Before introducing the fermentation broth into the column, the fermentation broth was passed through a T of the line where the CO2 bubbles were removed present in the feed through a ventilation hole.

With static internal components (without agitation), a difference in performance was observed when using fermentation broth extracted during the period of maximum gas evolution ("intermediate broth") in comparison with the broth extracted towards the end of the fermentation ("final broth"). With the use of the intermediate broth, a liquid throughput speed of 570 lpm / m2 (14 gpm / ft2) was achieved. The maximum yield (before the column flood) of the final broth was less than 367 lpm / m2 (9 gpm / ft2). There were notable differences in the size and appearance of the aqueous droplets. The droplet size of the aqueous phase was visibly higher in the final broth as compared to the intermediate broth.

Example 4 Effect of the concentration of isobutanol on the extraction efficiency of the column During a typical fermentation process, product levels change over time. This dynamic change of concentration can affect mass transfer in an extraction process.

In order to show the effect of the isobutanol concentration, a Karr® 2.5 cm (1") diameter glass extraction column (Koch Modular Process Systems, Paramus, NJ), equipped with internal stainless steel components, was used. , to process the fermentation broth from a fermentation containing approximately 3 g / 1 of isobutanol.The fermentation broth formed the continuous phase in the extractor, while the organic extractant (COFA) passed through the column in the form of droplets. Although, essentially, the production of CO2 had ceased, the fermentation broth was passed through a T of the line where the CO2 bubbles present in the feed were removed before the feed entered the extraction column.

Samples of the feed and outgoing streams were analyzed by liquid chromatography (LC) or gas chromatography (GC) to detect isobutanol. The results are shown in Table 4. Mass balances were performed and the height of an equilibrium transfer stage (HETS) was calculated by using the Kremser equations. The HETS values for the two data points in the "as is" fermentation broth were 3.0 and 4.0 meters (10 and 13 feet).

Then, isobutanol was added to the fermentation broth to bring the concentration to 20 g / 1. An extraction test was carried out and, from the data, a HETS value of 5.5 meters (18 feet) was reached. This value was approximately 50% higher than the values obtained in the raw broth and is in agreement with the data obtained with the use of light vinasse to which was added approximately 20 g / 1 of isobutanol (see Figure fifteen) .

Table 4 fifteen Example 5 ISPR with the use of an external extraction column Fermentation broth from a fermentation of isobutanol (10-liter scale) was circulated through a Karr® table column of 0.63 cm (5/8") in diameter.The extracting solvent (COFA) was reclimated from a tank of Extractant up to the Karr® column An experiment was carried out with a control fermentation, in which a volume of COFA was added to the fermenter for the continuous extraction of isobutanol from the fermentation broth.

During the fermentation, two experiments were performed with the Karr® column. The first experiment was at the time point of 4 to 7 hours of fermentation and the second experiment was at the time point of 22 to 33 hours of fermentation. In both fermentations, parameters such as p02 and pH were monitored. The p02 measurement was lower in the experiment in which the Karr® column was used, compared to the control experiment that did not use the Karr® column. The absolute pH values were similar in the Karr® column and the control, but the pH profiles were different in the two experiments. In the experiment with the Karr® column, the pH reached its peak in the initial stages, then remained constant and then reached the maximum again, compared to a gradual maximum for control.

In the Karr® column, two aliquots of extraction solvent (1.8 liters each) were analyzed. Samples were taken from each aliquot and the content of isobutanol was analyzed. The amount of isobutanol produced in the fermentation with the Karr® column was comparable to that produced in the control fermentation. The fermentation using the Karr® column produced a total of 82.4 grams of isobutanol: about 34 grams were in 3.6 liters of the organic phase and 48 grams in the aqueous phase. The control (30% by volume of organic phase added to the fermentor) produced 90 g / 1, 60 grams in 3 liters of the organic phase and 30 grams in the aqueous phase. The concentration of isobutanol in the aqueous phase was lower in the control due to the presence of COFA in the control fermenter from time zero, compared to a non-zero extraction start in the experiment with the Karr® column. In the Karr® column at 22 hours, isobutanol was extracted from the fermentor more rapidly than was being produced. Glucose profiles were generated for the control and the Karr® column. The profiles were similar, indicating that cell growth and metabolism were comparable. The results are shown in Figures 16A and 16B. The brackets indicate the temporary points (4 to 7 hours and 22 to 33 hours) in which the Karr® column was in operation.

Example 6 ISPR through the use of a mixer-settler An external mixer-settler system was used for the continuous removal of isobutanol from an active fermentation broth containing a microorganism that produced isobutanol (ie, an isobutanologen). The study used approximately 100 liters of fermentation broth inoculated with an isobutanol producing microorganism (ie, an isobutanologen). The content of the thermistor was recirculated from the thermistor by the extractor system of the mixer-settler. The extractant, which comprised COFA distillates that did not contain isobutanol, was circulated only once.

Two static mixers were evaluated. Most of the test used a Kenics® static stainless steel mixer with a diameter of 1.3 cm (¾ ") with 36 mixing elements.At the 12th and 24th hours of the experiment, a plastic mixer was used (StaMixCo HT- 11-12.6-24, StaMixCo LLC, Brooklyn, NY) The fermentation broth and the COFAs were introduced on the opposite sides of a T, from which the mixture flowed through the static mixer.The outgoing material of the static mixer was introduced. In the settler, the settler was made with a five-liter glass tank.

A dip tube passed through the top of the settler, near the perimeter and extended to approximately half the settler. The organic phase was removed by means of a port in the upper part of the settler, while the fermentation broth was removed from the bottom of the settler. The settler was equipped with an agitator that provided a gentle mixing to the aqueous-organic interface, to help uncouple the two liquid phases and, thus, minimize the accumulation of solids at the interface. The data collected during the experiment is presented in Table 5, and Figure 17 shows the isobutanol removal rates that were achieved during the course of the fermentation. As can be seen from the data, the isobutanol levels in the aqueous broth remained relatively constant, indicating that the isobutanol was removed from the fermentation broth at approximately the same rate at which it was produced. With reference to Table 5, Elapsed time is the time from the beginning of fermentation, Flow AQ is the flow of the aqueous feed, Flow ORG is the flow of organic feed, iB in feed AQ is isobutanol in the aqueous feed, and iB in ORG product is isobutanol in the product organic enriched Table 5 * A: Kenics® stainless steel mixer, from , 32 elements B: StaMixCo plastic mixer HT-11-12.6-24 Example 7 Measurements online, on line and in real time A tempered stream prepared from maize raw material was conducted to a three-phase centrifuge to generate three streams: temper, corn oil and wet cake. Process measurements were used in line or at the foot of the line, for example, to improve the recovery of starch / sugars and the quality of the corn oil, as well as to maximize the amount of starch / sugars extracted from the wet cake. Real-time measurements were used, for example, to control the addition of countercurrent water, cooking water or water to suspension tanks to maintain a target concentration value of starch / sugar. The amount of starch / sugar extracted from the wet cake was maximized by using a minimum amount of added water and reducing the hydraulic load in the three-stage centrifuge.

The samples of corn temper were analyzed by infrared spectroscopy with Fourier transform (FTIR), with a total attenuated reflectance probe (ATR, for its acronym in English) of diamond crystal, which allows measurements in the presence of solids. The FTIR was calibrated by collecting spectra from standard samples in which total starch / sugar determinations had been made using HPLC. The HPLC data was used to create a multivariate partial least squares model (PLS) for the FTIR. FTIR spectra were collected and a total concentration of starch was generated. Figure 18 illustrates the range of starch concentrations used to calibrate the FTIR.

The corn temper with an average starting concentration of 250 g / 1 was introduced in a three-phase centrifuge. The wet cake obtained was resuspended and the concentration of starch was measured in two samples: 80 g / 1 and 70 g / 1. This suspension was separated by the use of a three-phase centrifuge and the wet cake was resuspended. It was determined that the starch concentration of this suspension was 28.9 g / 1. The results are shown in Figure 19. These measurements were used to determine the correct amount of water to resuspend the wet cake at each stage. The optimization of the water addition maximized the concentration of starch and minimized the hydraulic load in the separation stage. The moisture content of the wet cake was measured by near infrared spectroscopy (NIR).

The quality of the corn oil was monitored in real time and the data was used to monitor the variables of the three-phase centrifuge (for example, feed speed, g-forces, input flow rate, travel speed). The quality of the corn oil generated by the three-phase centrifuge was measured by monitoring the water concentration incorporated in the corn oil during the separation. To collect the corn oil spectra as it left the three-phase centrifuge, FTIR was used with a diamond ATR probe. He Water detection limit with the use of the diamond ATR probe method was approximately 500 ppm. If a flow cell having a longer effective path length is used, lower detection limits are achieved.

Figure 20 contains a series of infrared spectra of corn oils containing a range of water concentrations in excess of concentrations of percentage levels less than hundreds of ppm. The water concentration was determined with the use of the -OH stretch region between 3700 cm-1 and 3050 cm-1. The data indicated that a process FTIR can be used to generate real-time water concentrations in oil data. The real-time water concentration data can be used to control the process variables of the three-phase centrifuge (eg, feed rate, g-forces, input flow, travel speed). The operation of the three-stage centrifuge can be controlled to produce the highest quality corn oil or to maximize the yield, without exceeding a target value for water.

The monitoring of the extractant in real time is used to detect and monitor the thermal decomposition of the extractant. The detection of these products of thermal decomposition in real time is used to trigger the recovery of the extractant or to purge the contaminated extractant from the process.

Figure 21 is an example of the real-time measurement of COFA enriched in isobutanol. The data was collected with a ReactIR ™ 247 from Metter-Toledo, by using a diamond ATR probe for samples in a flow cell. The COFA stream was collected from the outlet of a Karr® column 2.5 centimeters (1 inch) in diameter and pumped to the FTIR with the use of a peristaltic pump. The FTIR was calibrated through the creation of COFA standards enriched with isobutanol and the generation of a multivariate PLS model. Example 8 Analysis of the size of the droplet This example describes the analysis of the droplets of extractant liquid after conducting a process stream containing fermentation broth and extractant (COFA) to a static mixer. A PVM® probe (Mettler-Toledo, LLC, Columbus OH) was inserted into the process stream approximately 24 hours after the process stream left the static mixer. The PVM® probe was used to collect images every two minutes during a fermentation experiment. The images showed the presence of COFA droplets, whose size varied from 50 to 80 mm in diameter, and CO2 bubbles, whose size varied from 200 to 400 pm in diameter. Monitoring the droplet size in the process stream containing fermentation broth and COFA after the static mixer is used to ensure that the droplets remain below a particular average diameter, to ensure a good mass transfer of isobutanol into the COFA droplets The PVM® probe was also used to take pictures of the COFA droplets in the depleted broth stream before returning the current to the fermenter. The detection of COFA droplets in this stream is an indication of the amount of COFA that returns to the fermenter. The PVM® probe was used to collect, every two minutes, an image of the current during a fermentation. Unlike the current leaving the static mixer, the depleted broth stream had fewer droplets and smaller ones (10-40 pm). These measurements show the feasibility of using process images to monitor the amount of COFA that returns to the fermenter.

Real-time average droplet size data from both sample locations are used to monitor the phase separation of the fermentation broth and COFAs. An increase in the concentration or amount of small droplets of COFA detected in the recirculation stream of the depleted fermentation broth (after the extraction of isobutanol) may be an indicator that the phase separation of the fermentation broth has been degraded and the COFA and too many COFAs come out of the extractor. To improve the quality of phase separation and reduce the amount or concentration of COFA droplets returning to the In the impoverished broth stream, the average droplet size of COFA is increased after passing through the static mixer.

Other process variables that can affect the average COFA droplet size include the concentration of polysaccharides in the fermentation broth, the proportion of fermentation broth to COFA and the total flow rate in the static mixer. As the fermentation progresses, the proportions of the flow and / or fermentation broth to COFA can be changed so that the average droplet size of COFA remains constant.

Example 9 Exhaust design This example describes a method for designing a large-scale extraction unit. To calculate the size of the extractor unit on a large scale, data from a pilot scale extraction are used. The effects of flow, agitation speed and presence or absence of internal components in the phase separation of the extractor unit currents are determined from a pilot scale extraction. The total flow and the proportion of flow of fermentation broth to extractant flow is varied to a fixed temperature during the course of the fermentation and the conditions at which the phase separation is interrupted are observed. The maximum flow is recorded to the extractor unit that can reached per square foot of flow surface area to the extractor unit. To determine the flow per unit area, the following equation is used: - (Equation 1) U = flow per unit area (gallons / minute / square foot) F = total flow of the fermentation broth and extractant to the extractor unit (gallons / minute) A = area in cross section in the flow direction (square feet) for an extraction column this is given by pR 2 4 D = diameter of the column (feet).

The diameter of a large-scale extractor unit is calculated by the expected flow of the fermentation and extractant broth to the extractor unit by using the following equation: (Equation 2) Full-scale = total flow of the fermentation broth and extractant to the large-scale extractor (gallons / minute). The height of the pilot scale extractor unit is it measures with different flow regimes, which include different flow rates, with and without internal components present, different agitation speeds and different concentrations of the alcoholic product. With these data, the number of theoretical steps achieved by the height of the extractor unit is calculated by using the Kremser equation (Seader and Henlcy, Separation Process Principles, 2nd edition, John Wiley &Sons, 2006, pp. 358-359): - (Equation 3) E = extraction factor = Facade = flow of the broth to the extractor unit (gallons / minute) Fextractant = extractant flow to the extractor unit (gallons / minute) m = coefficient of partition of the alcoholic product in the phases of the fermentation broth and the extractant (g / 1 per g / 1) Xf = concentration of alcoholic product in the feed of fermentation broth (g / 1) Xn = concentration of alcoholic product in the broth of fermentation leaving the extractor unit (g / 1) Ys = concentration of alcoholic product in the extractant that enters the extractor unit (g / 1) n = number of theoretical stages reached by the height of the extractor unit Equation 3 is only valid when E ¹ 1.

The height of a theoretical stage of the extractor unit is given by the height of the extraction column used in the pilot scale extraction divided by the number of theoretical stages reached in a given experiment. The number of theoretical steps required to achieve large-scale separation is calculated by using the large-scale expected operating conditions in Equation 4: _ (Equation 4) where 'indicates the condition of the extractor unit on a large scale.

The product of the number of theoretical stages and the height of a theoretical stage measured under similar flow conditions provides a calculation of the total height of the large-scale extractor unit. The flows and concentrations expected in a large extraction unit Scale is calculated by using a dynamic fermentation model (eg, Daugulis, et al., Biotech, Bioeng., 27: 1345-1356, 1985).

While the present disclosure has described various embodiments of the present invention, it should be construed that they have been presented as an example only and not in a limiting manner. It will be apparent to persons with experience in the pertinent technique that various changes in the form and details of this can be made without departing from the spirit and scope of the invention. Therefore, the breadth and scope of the present invention should not be limited by the illustrative embodiments described above, but should be defined only in accordance with the following claims and their equivalents.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. A method for recovering an alcoholic product from a fermentation broth, characterized in that it comprises: providing a fermentation broth comprising a microorganism, wherein the microorganism produces an alcoholic product; contacting the fermentation broth with at least one extractant; Y recover the alcoholic product.

2. The method according to claim 1, characterized in that the contacting of the fermentation broth with at least one extractant occurs in the fermenter, an external unit or both.

3. The method according to claim 2, characterized in that the external unit is an extractor.

4. The method according to claim 3, characterized in that the extractor is selected from siphon, decanter, centrifuge, gravity settler, phase splitter, mixer-settler, column extractor, centrifugal extractor, agitator, hydro-discharge, spray tower or combinations of these.

5. The method according to claim 1, characterized in that the extractant is selected from C7 to C22 alcohols, C7 to C22 fatty acids, C7 to C22 fatty acid esters, C7 to C22 fatty aldehydes, C7 to C22 fatty amides, and mixtures thereof.

6. The method according to claim 1, characterized in that the at least one extractant is selected from oleyl alcohol, behenyl alcohol, cetyl alcohol, lauryl alcohol, myristyl alcohol, stearyl alcohol, oleic acid, lauric acid, linoleic acid, linolenic acid, myristic, stearic acid, octanoic acid, decanoic acid, undecanoic acid, methyl myristate, methyl oleate, 1-nonanol, 1-decanol, 2-undecanol, 1-nonanal, 1-undecanol, undecanal, lauric aldehyde, 2-methylundecanal , oleamide, linoleamide, palmitamide, stearylamide, 2-ethyl-1-hexanol, 2-hexyl-1-decanol, 2-octyl-1-dodecanol, and mixtures thereof.

7. The method according to claim 6, characterized in that a hydrophilic solute is added to the fermentation broth.

8. The method according to claim 7, characterized in that the hydrophilic solute is selected from the group consisting of polyhydroxy compounds, polycarboxylic acids, polyol compounds, ionic salts, or mixtures thereof.

9. The method according to claim 1, characterized in that the contacting of the fermentation broth with at least one extractant takes place in two or more external units.

10. The method according to claim 1, characterized in that the contacting of the fermentation broth with at least one extractant takes place in two or more fermenters.

11. The method according to claim 10, characterized in that the burners comprise internal components or devices for improving phase separation.

12. The method according to claim 11, characterized in that the internal components or devices are selected from the group consisting of coalescing agents, baffle plates, perforated plates, wells, sheet separators, cones or combinations thereof.

13. The method according to claim 1, characterized in that real-time measurements are used to monitor the extraction of the alcoholic product.

14. The method according to claim 13, characterized in that the extraction of the alcoholic product is monitored by means of real-time measurements of the phase separation.

15. The method according to claim 14, characterized in that the phase separation is monitored by measuring the phase separation rate, the droplet size of the extractant and / or the composition of the fermentation broth.

16. The method according to claim 15, characterized in that the phase separation is monitored by conductivity measurements, dielectric measurements, viscoelastic measurements, or ultrasonic measurements.

17. The method according to claim 1, characterized in that the provision of a fermentation broth comprising a microorganism takes place in two or more fermenters.

18. The method according to claim 1, characterized in that the alcohol product is selected from ethanol, propanol, butanol, pentanol, hexanol, and fusel alcohols.

19. The method according to claim 1, characterized in that the microorganism comprises a biosynthetic route of butanol.

20. The method according to claim 19, characterized in that the biosynthetic route of butanol is a biosynthetic route of 1-butanol, a biosynthetic route of 2-butanol or a biosynthetic route of isobutanol.

21. The method according to claim 19, characterized in that the microorganism is a recombinant microorganism.

22. The method according to claim 1, characterized in that it also comprises the steps of: providing a suspension of raw material, comprising a source of fermentable carbon, undissolved solids, oil and water; separating the suspension of raw material by which (i) an aqueous solution comprising a fermentable carbon source is formed, (ii) a wet cake comprising solids and (iii) an oil; Y add the aqueous solution to the fermentation broth.

23. The method according to claim 22, characterized in that the oil is hydrolyzed to form fatty acids.

24. The method according to claim 23, characterized in that the fermentation broth is brought into contact with the fatty acids.

25. The method according to claim 23, characterized in that the oil is hydrolyzed by an enzyme.

26. The method according to claim 25, characterized in that the enzyme is one or more lipases or phospholipases.

27. The method according to claim 22, characterized in that the suspension of raw material is generated by hydrolysis of the raw material.

28. The method according to claim 27, characterized in that the raw material is selected from rye, wheat, corn, sugar cane, barley, cellulose or lignocellulosic material, or combinations thereof.

29. The method according to claim 22, characterized in that the suspension of raw material is separated by centrifugation in bowl decanter, three-phase centrifugation, disk-spin centrifugation, filtration centrifugation, decanter centrifugation, filtration, vacuum filtration, band filter, pressure filtration, filtration through the use of a screen, screen separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, vortex separator, or combination of these.

30. The method according to claim 22, characterized in that the separation of the raw material is a one-stage process.

31. The method according to claim 22, characterized in that the wet cake is combined with the aqueous solution.

32. The method according to claim 22, characterized in that it further comprises contacting the aqueous solution with a catalyst to convert the oil of the aqueous solution into fatty acids.

33. The method according to claim 32, characterized in that the aqueous solutions and the fatty acids are added to the fermentation broth.

34. The method according to claim 32, characterized in that the catalyst is deactivated.

35. A system characterized in that it comprises: One or more workers who comprise: an entry to receive the suspension of raw material; Y an outlet for discharging the fermentation broth comprising alcoholic product; Y one or more extractors comprising: a first entry to receive the fermentation broth; a second entry to receive the extractant; a first outlet to discharge an impoverished fermentation broth; Y a second outlet to discharge an enriched extractant.

36. The system in accordance with the claim 35, characterized in that it also comprises: one or more liquefaction units; one or more separation means; Y optionally, one or more washing systems.

37. The system according to claim 36, characterized in that the separation means are selected from decanter centrifugation, three-phase centrifugation, disk-stack centrifugation, filtration centrifugation, decanter centrifugation, filtration, vacuum filtration, band filter. , pressure filtration, membrane filtration, microfiltration, filtration through the use of a screen, sieve separation, grid separation, porous grid separation, flotation, hydrocyclone, filter press, screw press, gravity settler, separator vortex, and combinations of these

38. The system according to claim 35, characterized in that the system comprises a loop measuring device.

39. The system according to claim 38, characterized in that the loop measuring devices are selected from particle size analyzers, infrared spectroscopes by Fourier transform, near infrared spectroscopes, Raman spectroscopes, high performance liquid chromatography. pressure, viscometers, densitometers, tensiometers, droplet size analyzers, pH meters, dissolved oxygen probes, or combinations of these.