The invention relates to the fields of industrial microbiology and alcohol production including production of yeast products with features suitable for transport, storage, and utilization in fermentation
The present disclosure is directed to methods, approaches, devices, equipment, and systems for minimizing or reducing contamination in facilities implementing fermentation or distillation processes. In embodiments, the facility is a biofuel plant that produces fermentation product such as product alcohol like butanol. In some embodiments, the methods, approaches, devices, equipment, and systems are operable to implement clean in place contamination (CIP) mitigation techniques that can also include sterilize in place (SIP) mitigation techniques to decontaminate equipment including surfaces of the equipment that come in contact with materials used in the production of product alcohols. Other cleaning and contamination minimizing techniques are also described.
B01D 3/00 - Distillation or related exchange processes in which liquids are contacted with gaseous media, e.g. stripping
B08B 3/04 - Cleaning involving contact with liquid
C11D 1/68 - AlcoholsOxidation products of paraffin wax, other than acids
C12M 1/40 - Apparatus specially designed for the use of free, immobilised, or carrier-bound enzymes, e.g. apparatus containing a fluidised bed of immobilised enzymes
3.
EXPRESSION OF A HAP TRANSCRIPTIONAL COMPLEX SUBUNIT
The invention relates, for example, to recombinant yeast cells for differential gene expression during the propagation and production phases of a fermentation-based production process, as well as methods for using the same. The invention is directed to recombinant yeast cells that comprise a recombinant polynucleotide encoding a gene for a subunit of the HAP transcriptional complex. The cells further comprise promoter sequences that provide differential expression in the propagation vs. production phases of a process, as well as methods for using the same. In embodiments, the cells have increased growth rate, tcrit and/or biomass production. In other embodiments, the cells produce a fermentation product.
Provided herein are recombinant yeast cells comprising an engineered pyruvate utilizing biosynthetic pathway and further comprising a cell membrane with an altered lipid profile. In some embodiments the recombinant yeast cell has an increased tolerance to butanol as compared to a recombinant yeast cell that does not comprise an altered lipid profile. In some embodiments the altered lipid profile comprises an increase in the concentration of cyclopropane fatty acid as compared to a microorganism that lacks the cell membrane with an altered lipid profile. In some embodiments the altered lipid profile comprises an increase in the fatty acid concentration ratio.
A process for recovering alcohol from a fatty acid and/or diacid alcohol ester using water in an equal or lesser amount than oil on a mass basis. The process uses multiple reactors with separation of the alcohol product of hydrolysis between successive reactors. The use of low amounts of water allows recovery of the alcohol with a lower evaporation requirement, thus making a more energy efficient process.
Provided herein are recombinant yeast host cells and methods for their use for production of fermentation products from a pyruvate utilizing pathway. The yeast host cells provided herein comprise at least one genetic modification in a pyruvate decarboxylase gene and at least one genetic modification in an endogenous cell wall protein, which confers resistance to butanol and increased glucose utilization.
The invention relates to the use of various post-transcriptional strategies for differential expression of genes during the propagation and production phases of a fermentation process. Aspects of the invention relate to recombinant host cells that comprise identified promoter nucleic acid sequences and methods for producing fermentation products employing the same.
The invention relates to recombinant host cells that comprise controlled biocatalyst polypeptides and methods for producing fermentation products employing the same. In some embodiments, various controlled polypeptides and control cues are employed during the propagation and production phases of a fermentation process.
Provided herein are processes for producing an improved culture of cells comprising an engineered butanol biosynthetic pathway. The processes comprise (a) providing a cell culture of recombinant microorganisms comprising an engineered butanol biosynthetic pathway, wherein the engineered butanol biosynthetic pathway is minimall or not activated; and (b) growing the culture of recombinant microorganisms under adaptive conditions whereby pathway activation is increased to produce an improved cell culture and whereby said improved cell culture is capable of continuing to grow in fermentation.
The present invention relates to the fermentative production of product alcohols including ethanol and butanol, and related co-products, and processes for improving alcohol fermentation employing in situ product removal methods.
The present invention relates to processes and systems for the production of fermentative products such as ethanol and butanol. The present invention also provides methods for separating feed stream components for improved biomass processing productivity.
The present invention relates to processes and systems for the production of fermentative alcohols such as ethanol and butanol. The present invention also provides methods for separating feed stream components for improved biomass processing and productivity.
Methods of screening for dihydroxy-acid dehydratase (DHAD) variants that display increased DHAD activity are disclosed, along with DHAD variants identified by these methods. Such enzymes can result in increased production of compounds from DHAD requiring biosynthetic pathways. Also disclosed are isolated nucleic acids encoding the DHAD variants, recombinant host cells comprising the isolated nucleic acid molecules, and methods of producing butanol.
Provided herein are methods of maximizing biomass production and butanol (e.g., isobutanol) yield during fermentation of recombinant microorganisms. Biomass production during a biomass growth phase and/or butanol production during a butanol production phase can be maximized by adjusting a manipulated variable, such as feed rate, in response to a controlled variable, such as respiratory quotient (RQ).
The invention relates to a method for producing butanol through microbial fermentation, in which the butanol product is removed by extraction into a water immiscible organic extractant during the fermentation. The invention also relates to a method for producing butanol through microbial fermentation, in which the butanol product is removed during the fermentation by extraction into a water-immiscible extractant composition. The invention further relates to compositions comprising a solution of butanol in a water immiscible organic extractant composition.
Provided herein are recombinant yeast host cells and methods for their use for production of fermentation products. Host cells provided herein comprise a pyruvate-utilizing pathway and a competitive growth advantage over other microorganisms in solution.
The invention relates to a method for producing butanol through microbial fermentation, in which the butanol product is removed by extraction into a water immiscible organic extractant composition during the fermentation. The invention also relates to a method for producing butanol through microbial fermentation, in which the butanol product is removed during the fermentation by extraction into a water-immiscible extractant composition comprising a first solvent having a butanol partition coefficient of at least 3, and a second solvent having a butanol partition coefficient, wherein the butanol partition coefficient of the first solvent is higher than the butanol partition coefficient of the second solvent. Optionally, the first solvent has a higher concentration of hydrogen bonding sites than the second solvent. The invention further relates to a composition including butanol in the water immiscible organic extractant composition described above.
Provided herein are methods for recovering butanol from a fermentation medium. The methods comprise providing a fermentation medium comprising butanol, water, and a recombinant microorganism comprising a butanol biosynthetic pathway, wherein the recombinant microorganism produces butanol; contacting the fermentation medium with a water immiscible organic extractant composition comprising a dry solvent to form a butanol-containing organic phase and an aqueous phase; and recovering the butanol from the butanol-containing organic phase.
Provided are processes for recovering thermal energy and utilizing the recovered thermal energy as a heat source. The processes comprise distilling a butanol and water composition in a distillation unit, whereby the distillation produces a vapor comprising butanol and water at a pressure P1; compressing the vapor to a second pressure P2, wherein P2 is greater than P1; and condensing the vapor to a liquid, whereby condensing the vapor allows for the recovery of thermal energy, wherein the recovered thermal energy is used as a heat source.
The invention relates to the development of microorganisms capable of producing fermentation products via an engineered pathway in the microorganisms. The invention also relates to microorganisms with improved cell productivity and methods to improve productivity of a microorganism.
Provided herein are processes for adjusting a fermentation medium to reduce the activity of one or more carboxylic acids. The processes comprise (a) providing a recombinant microorganism comprising an engineered butanol biosynthetic pathway, (b) contacting the recombinant microorganism with a fermentation medium whereby butanol is produced and wherein the fermentation medium comprises one or more carboxylic acids, and (c) adjusting the fermentation medium to reduce the activity of the one or more carboxylic acids. Also provided are processes for reducing the activity of one or more carboxylic acids in a feed. The processes comprise (a) providing a feed from a fermentation vessel, wherein the feed comprises a composition produced by a recombinant microorganism comprising an engineered butanol biosynthetic pathway, wherein the composition comprises butanol, water, and one or more carboxylic acids; and (b) adjusting the feed, wherein adjusting the feed reduces the activity of the one or more carboxylic acids.
The present invention relates to processes and systems for the production of fermentation products such as alcohols. The present invention also provides methods for separating feed stream components for improved biomass processing and productivity.
Polypeptides having ketol-acid reductoisomerase activity are provided. Also disclosed are recombinant host cells comprising isobutanol biosynthetic pathways employing such polypeptides. Methods for producing isobutanol employing host cells comprising the polypeptides having ketol-acid reductoisomerase activity are also disclosed.
The invention relates to processes for the production of fermentation products such as alcohols including ethanol and butanol, and the development of microorganisms capable of producing fermentation products via an engineered pathway in the microorganisms.
Provided herein are processes comprising providing a fermentation medium comprising a fermentable carbon source, a recombinant microorganism comprising an engineered butanol biosynthetic pathway, and butanol; contacting said fermentation medium with an extractant composition comprising an effective amount of antioxidant or antioxidant-like compound, whereby at least a portion of the butanol in the fermentation medium partitions into the extractant; recovering at least a portion of the butanol and extractant composition from the fermentation medium; recycling the extractant composition recovered in (c) one or more times in the fermentation medium; optionally adjusting the effective amount of antioxidant or antioxidant-like compound in the extractant composition, whereby the rate of oxidation or effet of oxidized products in the recycled extractant composition is substantially reduced and/or avoided such that the extractant composition may be recycled.
Provided herein are processes and microorganisms which utilize both protein hydrolysates and carbohydrates from biomass feedstocks to produce renewable hydrocarbon compositions. Advantages of the disclosed methods may be recognized in fuel blends comprising such hydrocarbon compositions.
C12N 15/63 - Introduction of foreign genetic material using vectorsVectorsUse of hosts thereforRegulation of expression
F02D 19/08 - Controlling engines characterised by their use of non-liquid fuels, pluralities of fuels, or non-fuel substances added to the combustible mixtures peculiar to engines working with pluralities of fuels, e.g. alternatively with light and heavy fuel oil, other than engines indifferent to the fuel consumed simultaneously using pluralities of fuels
27.
PROCESSES AND SYSTEMS FOR THE PRODUCTION OF FERMENTATION PRODUCTS
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.
The invention relates to the development of microorganisms capable of producing fermentation products via an engineered pathway in the microorganisms. The invention also relates to microorganisms with improved cell viability and methods to improve cell viability and cell productivity of a microorganism.
C12N 1/00 - Microorganisms, e.g. protozoaCompositions thereofProcesses of propagating, maintaining or preserving microorganisms or compositions thereofProcesses of preparing or isolating a composition containing a microorganismCulture media therefor
29.
PROCESSES AND SYSTEMS FOR THE PRODUCTION OF FERMENTATION PRODUCTS
The present invention relates to processes and systems for the production of fermentation products such as alcohols. The present invention also provides methods for separating feed stream components for improved biomass processing and productivity.
Provided are methods for removing one or more components from a butanol based composition. The methods comprise providing a butanol based composition comprising one or more components, targeting at least one component or a combination thereof for reduction, and processing said butanol based composition such that the at least one targeted component is substantially removed. The butanol based composition can, for example, be bio-produced.
Provided herein are polypeptides having ketol-acid reductoisomerase activity as well as microbial host cells comprising such polypeptides. Polypeptides provided herein may be used in biosynthetic pathways, including, but not limited to, isobutanol biosynthetic pathways.
The present invention relates to processes for recovering butanol produced in a fermentative process using, for example, an ethanol production plant which has been reversibly retrofitted for butanol production, that is, the ethanol production plant may be converted for butanol production, but can also revert to an ethanol production. The present invention also relates to processes for recovering butanol produced in a fermentative process in a butanol production plant that may be converted to ethanol production plant.
The invention relates to the fields of industrial microbiology and alcohol production. More specifically, the invention relates to improved production of butanol isomers by recombinant microorganisms containing an engineered butanol pathway and disrupted activity of the genes in pathways for the production of by-products during the fermentation when the microorganisms are grown in a fermentation medium containing acetate. In embodiments, recombinant microorganisms have an increased growth rate in a fermentation medium containing acetate as a C2 supplement.
This invention relates to corrosion inhibitor additive combinations giving long acting performance in oxygenated gasoline blends comprising either low carbon number (< 3) or high carbon number (greater than or equal to 4) alcohols or mixtures thereof and adapted for use in fuel delivery systems and internal combustion engines. The invention also is concerned with a process for conferring anti-corrosion properties to oxygenates in gasoline fuel mixtures wherein the oxygenate comprises biologically-derived butanol.
The invention relates to the fields of industrial microbiology and alcohol production. The invention also relates to the development of a microorganism capable of producing fermentation products via an engineered pathway, and uses of the microorganism. A process for producing butanol comprising the following steps: (a) providing a butanologen, (b) contacting the butanologen with one or more carbon substrates under conditions wherein butanol is produced at an effective yield; (c) collecting the butanologen; (d) recovering butanol at a concentration of at least about 6 g/L; (e) contacting the collected butanologen of (c) with one or more carbon substrates under conditions wherein butanol is produced at an effective yield and wherein the effective yield is at least about 90% of the effective yield of (b); (f) repeating steps (c)-(e); and, optionally exposing the collected butanologen of (c) to conditions of pH less than or equal to about 2.0 for at least about 1 hour in the presence of at least about 0.3% butanol. The invention also relates to the methods to improve cell viability and productivity and the use of recycling and acid washing to increase the yield of fermentation products.
The invention relates to suitable screening strategies for evaluating various candidate promoters for differential gene expression during the propagation and production phases of a fermentation process. The invention also relates to recombinant host cells that comprise identified promoter nucleic acid sequences and methods for producing fermentation products employing the same.
The present invention relates to processes to remove butanol and other product alcohols from a fermentation broth employing vacuum vaporization. The method for removing a product alcohol from a fermentation broth comprises the following steps: (a) the vaporisation of a fermentation broth or a portion thereof to form one or more vapor streams, wherein vaporizing comprises: (i) vaporizing the fermentation broth or a portion thereof by one or more pre-flashes; and (ii) vaporizing the fermentation broth or a portion thereof by a flash; wherein the vapor streams comprise one or more components selected from product alcohol, water, and carbon dioxide; and (b) recovering the product alcohol from the one or more vapor streams or a portion thereof.
Systems and processes for the production of fuel and fuel blends involve the production of fuels for blending with one or more alcohols such as ethanol and/or butanol. A method for producing a fuel blend includes blending a light distillate product from an oil refinery with butanol. The fuel blending can be at the oil refinery.
The invention relates to butanol compositions for fuel blending and fuel blends comprising such compositions. The compositions and fuel blends of the invention have desirable performance characteristics and can serve as alternatives to ethanol-containing fuel blends. The invention also relates to methods for producing such butanol compositions and fuel blends.
Provided herein are polypeptides and polynucleotides encoding such polypeptides which have ketoisovalerate decarboxylase activity Also provided are recombinant host cells comprising such polypeptides and polynucleotides and methods of converting a-ketoisovalerate to isobutyraldehyde or methods of producing isobutanol by employing the polypeptides disclosed.
The invention relates generally to the field of industrial microbiology and butanol production from sources of 5-carbon sugars such as lignocellulosic hydrolysates. More specifically, the invention relates to the use of an xylulose or xylulose-5 -phosphate-producing enzyme and micro-aerobic or anaerobic conditions to increase butanol production from such sugars and recovery of said butanol through ins situ product recovery methods.
The present invention includes methods of generating co-products for animal feed and compositions useful as co-products for animal feed derived from biofuel production processes. More specifically, the invention includes co-products for animal feed from at least one process feedstream, such as fatty acids from oil hydrolysis, lipids from evaporation of thin stillage, syrup, distillers grains, distillers grains and solubles, solids from a mash before fermentation, and solids from a whole stillage after fermentation.
Disclosed herein are methods of producing alcohol esters during a fermentation by providing alcohol-producing microorganisms which further comprise an engineered polynucleotide encoding a polypeptide having lipase activity.
C12N 1/38 - Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factorsStimulation of growth by removal of a chemical compound
The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells.
Fatty acids derived from biomass at a step in a fermentation process can be added to a fermentation medium comprising a recombinant microorganism that produces a product alcohol. At least one of growth rate and fermentable carbon consumption of the microorganism is greater in the presence of the fatty acids than the growth rate and the fermentable carbon consumption of the microorganism in the absence of the fatty acids. The addition of the fatty acids can increase glucose consumption, and can improve microorganism biomass production (cell growth/density) and growth rate, thereby reducing production time and increasing productivity of the fermentation process.
In an alcohol fermentation process, oil derived from biomass is hydrolyzed into an extractant available for in situ removal of a product alcohol such as butanol from a fermentation broth. The glycerides in the oil can be catalytically (e.g., enzymatically) hydrolyzed into free fatty acids, which form a fermentation product extractant having a partition coefficient for a product alcohol greater than a partition coefficient of the oil of the biomass for the product alcohol. Oil derived from a feedstock of an alcohol fermentation process can be hydrolyzed by contacting the feedstock including the oil with one or more enzymes whereby at least a portion of the oil is hydrolyzed into free fatty acids forming a fermentation product extractant, or the oil can be separated from the feedstock prior to the feedstock being fed to a fermentation vessel, and the separated oil can be contacted with the enzymes to form the fermentation product extractant. The fermentation product extractant can be contacted with a fermentation broth for in situ removal of a product alcohol.
In an alcohol fermentation process, oil derived from biomass is chemically converted into an extractant available for in situ removal of a product alcohol such as butanol from a fermentation broth. The glycerides in the oil can be chemically converted into a reaction product, such as fatty acids, fatty alcohols, fatty amides, fatty acid methyl esters, fatty acid glycol esters, and hydroxylated triglycerides, and mixtures thereof, which forms a fermentation product extractant having a partition coefficient for a product alcohol greater than a partition coefficient of the oil of the biomass for the product alcohol. Oil derived from a feedstock of an alcohol fermentation process can be chemically converting into the fermentation product extractant. The oil can be separated from the feedstock prior to the feedstock being fed to a fermentation vessel, and the separated oil can be chemically converted to a fermentation product extractant, which can then contacted with a fermentation product comprising a product alcohol, whereby the product alcohol is separated from the fermentation product.
A method and system for efficiently producing a fermentative product alcohol such as butanol utilizing in situ product extraction are provided. The efficiency is obtained through separating undissolved solids after liquefying a given feedstock to create a feedstock and prior to fermentation, for example, through centrifugation. Removal of the undissolved solids avoids problems associated with having the undissolved solids present during in situ production extraction, and thereby increases the efficiency of the alcohol production.
The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
High cell density cultures of yeast were found to have higher tolerance for butanol in the medium. The high cell density yeast cultures had greater survival and higher glucose utilization than cultures with low cell densities. Production of butanol using yeast in high cell density cultures is thus beneficial for improving butanol production.
A gasoline blend and a method for producing a gasoline blend containing low concentrations of a butanol isomer and having good cold start and warm-up driveability characteristics are disclosed.
Gasoline blends and methods for producing gasoline blends containing high concentrations of a butanol isomer and having good cold start and warm-up driveability characteristics are disclosed.
An alcohol fermentation process and composition that includes production of alcohol esters by esterification of product alcohol in a fermentation medium with a carboxylic acid (e.g., fatty acid) and a catalyst (e.g., lipase) capable of esterifying the product alcohol, such as butanol, with the carboxylic acid to form the alcohol esters. The alcohol esters can be extracted from the fermentation medium, and the product alcohol recovered from the alcohol esters. The carboxylic acid can also serve as an extractant for removal of the alcohol esters from the fermentation medium.
The present invention is related to a recombinant host cell, in particular a yeast cell, comprising a dihydroxy-acid dehydratase polypeptide. The invention is also related to a recombinant host cell having increased specific activity of the dihydroxy-acid dehydratase polypeptide as a result of increased expression of the polypeptide, modulation of the Fe-S cluster biosynthesis of the cell, or a combination thereof. The present invention also includes methods of using the host cells, as well as, methods for identifying polypeptides that increase the flux in an Fe-S cluster biosynthesis pathway in a host cell.
A fermentation liquid feed including water and a product alcohol and optionally CO2 is at least partially vaporized such that a vapor stream is produced. The vapor stream is contacted with an absorption liquid under suitable conditions wherein an amount of the product alcohol is absorbed. The portion of the vapor stream that is absorbed can include an amount of each of the water, the product alcohol and optionally the CO2. The temperature at the onset of the absorption of the vapor stream into the absorption liquid can be greater than the temperature at the onset of condensation of the vapor stream in the absence of the absorption liquid. The product alcohol can be separated from the absorption liquid whereby the absorption liquid is regenerated. The absorption liquid can include a water soluble organic molecule such as an amine.
The invention relates to suitable candidate ADH enzymes for production of lower alkyl alcohols including isobutanol. The invention also relates to recombinant host cells that comprise such ADH enzymes and methods for producing lower alkyl alcohols in the same.
Systems and methods for separating an alcohol, and in particular butanol, from a fermented feed and concentrating thin stillage into syrup includes operation of one or more alcohol recovery distillation columns using the heat supplied by steam generated from concentration of the thin stillage in a multi-train, multi-effect evaporation system.
The invention relates to a recombinant host cell having (a) a modification in an endogenous polynucleotide encoding a polypeptide having dual-role hexokinase activity; (b) a heterologous polynucleotide encoding a polypeptide having hexose kinase activity; and optionally (c) a modification in an endogenous polynucleotide encoding a polypeptide having pyruvate decarboxylase activity. Additionally, the invention relates to methods of making and using such recombinant host cells including, for example, methods of increasing glucose consumption, methods of improving redox balance, and/or methods of increasing the production of a product of a pyruvate-utilizing pathway.
A method for producing butanol through microbial fermentation, in which the butanol product is removed during the fermentation by extraction into a water-immiscible organic extractant in the presence of at least one electrolyte at a concentration at least sufficient to increase the butanol partition coefficient relative to that in the presence of the salt concentration of the basal fermentation medium, is provided. The electrolyte may comprise a salt which dissociates in the fermentation medium, or in the aqueous phase of a biphasic fermentation medium, to form free ions. Also provided is a method and composition for recovering butanol from a fermentation medium.
A method is provided for producing butanol through microbial fermentation, in which the butanol product is removed during the fermentation by extraction into a water-immiscible organic extractant in the presence of at least one osmolyte at a concentration at least sufficient to increase the butanol partition coefficient relative to that in the presence of the osmolyte concentration of the basal fermentation medium and of an optional fermentable carbon source. The osmolyte may comprise a monosaccharide, a disaccharide, glycerol, sugarcane juice, molasses, polyethylene glycol, dextran, high fructose corn syrup, corn mash, starch, cellulose, and combinations thereof. Also provided is a method and composition for recovering butanol from a fermentation medium.
A process for recovering butanol from a mixture comprising a water-immiscible organic extractant, water, butanol, and optionally a non-condensable gas, is provided. The butanol is selected from 1-butanol, 2-butanol, isobutanol, and mixtures thereof. An overhead stream from a first distillation column is condensed to recover a mixed condensate. An entrainer is added to at least one appropriate process stream or vessel such that the mixed condensate comprises sufficient entrainer to provide phase separation of the organic and the aqueous phases to provide for recovery of the butanol.
A process for recovering butanol from a mixture comprising a water-immiscible organic extractant, water, butanol, and optionally a non-condensable gas, is provided. The butanol is selected from 1-butanol, 2-butanol, isobutanol, and mixtures thereof. The extractant comprises at least one solvent selected from the group consisting of C7 to C22 fatty alcohols, C7 to C22 fatty acids, esters of C7 to C22 fatty acids, C7 to C22 fatty aldehydes, and mixtures thereof.
Crabtree positive yeast cells that have endogenous expressed pyruvate decarboxylase genes inactivated and an engineered biosynthetic pathway utilizing pyruvate were found to have improved growth and product yield when glucose repression was reduced. These cells were able to grow in media containing a high glucose concentration.
A high flux in conversion of pyruvate to acetolactate was achieved in yeast through expression of acetolactate synthase in the cytosol in conjunction with reduction in pyruvate decarboxylase activity. Additional manipulations to improve flux to acetolactate are reduced pyruvate dehydrogenase activity and reduced glycerol-3-phosphate dehydrogenase activity. Production of compounds having acetolactate as an upstream intermediate benefit from the increased conversion of pruvate to acetolactate in the described strains.
An engineering method was developed to allow genetic modification and isolation of lactic acid bacteria cells that lack lactate dehydrogenase.and acetolactate decarboxylase activities. In cells with these modifications and an isobutanol biosynthetic pathway, improved production of isobutanol was observed.
Ketol-acid reductoisomerase enzymes have been identified that provide high effectiveness in vivo as a step in an isobutanol biosynthetic pathway in bacteria and in yeast. These KARIs are members of a clade identified through molecular phylogenetic analysis called the SLSL Clade.
A process for recovering butanol from a mixture comprising a water-immiscible organic extractant, water, butanol, and optionally a non-condensable gas, is provided. The butanol is selected from 1-butanol, isobutanol, and mixtures thereof. An overhead stream from a first distillation column is decanted into two liquid phases. The wet butanol phase is refined in a second distillation column; the aqueous phase is returned to the first distillation column. A portion of the wet butanol phase from the decanter is also returned to the first distillation column. The extractant comprises at least one solvent selected from the group consisting of C7 to C22 fatty alcohols, C7 to C22 fatty acids, esters of C7 to C22 fatty acids, C7 to C22 fatty aldehydes, and mixtures thereof.
A process for recovering butanol from a mixture comprising a water-immiscible organic extractant, water, butanol, and optionally a non-condensable gas, is provided. The butanol is selected from 1-butanol, isobutanol, and mixtures thereof. An overhead stream from a first distillation column is decanted into two liquid phases. The wet butanol phase is returned to the first distillation column as reflux. A bottom stream from the first distillation column is refined in a second distillation column to obtain a second overhead stream comprising butanol and a second bottoms stream comprising the extractant. The extractant comprises at least one solvent selected from the group consisting of C7 to C22 fatty alcohols, C7 to C22 fatty acids, esters of C7 to C22 fatty acids, C7 to C22 fatty aldehydes, and mixtures thereof.
A method for producing butanol through microbial fermentation, in which the butanol product is removed by extraction into a water-immiscible extractant composition comprising a first solvent and a second solvent, is provided. The first solvent is selected from the group consisting of C12 to C22 fatty alcohols, C12 to C22 fatty acids, esters of C12 to C22 fatty acids, C12 to C22 fatty aldehydes, C12 to C22 fatty amides and mixtures thereof. The second solvent is selected from the group consisting of C7 to C11 alcohols, C7 to C11carboxylic acids, esters of C7 to C11 carboxylic acids, C7 to C11 aldehydes, and mixtures thereof. Also provided is a method for recovering butanol from a fermentation medium.
Increasing tolerance to butanol in yeast has been accomplished by decreasing activity of Pdr5p encoded by an endogenous PDR5 gene. A deletion mutation of the PDR5 gene led to improved growth yield in the presence of butanol. Yeast cells with reduced Pdr5p activity, or other multidrug resistance ATP-binding cassette transporter protein activity encoded by CDR1 or BFR1, and a butanol biosynthetic pathway may be used for improved butanol production
[0070] A gasoline deposit control additive composition for use in a fuel comprising from about 70 to about 95 volume percent of hydrocarbons in the gasoline boiling range and from about 5 to about 30 volume percent of at least one alcohol, comprising the imine or tertiary amine product of the reaction between (a) at least one aldehyde or ketone or mixture thereof having the formula R16 CHO, R16 CH2 CHO, R17 (C=O) R18 or R17 CH2 (C=O) R18, wherein R16, R17, and R18 are the same or different and are each independently a straight or branched chain hydrocarbyl or aryl group that contains from 1 to 18 carbon atoms, and (b) a primary or secondary amine functionality.
C10L 1/228 - Organic compounds containing nitrogen containing at least one carbon-to-nitrogen double bond, e.g. guanidines, hydrazones, semicarbazones, iminesOrganic compounds containing nitrogen containing at least one carbon-to-nitrogen triple bond, e.g. nitriles
C10L 1/222 - Organic compounds containing nitrogen containing at least one carbon-to-nitrogen single bond
C10L 10/18 - Use of additives to fuels or fires for particular purposes use of detergents or dispersants for purposes not provided for in groups
C10L 1/2383 - Polyamines or polyimines, or derivatives thereof
C10L 1/232 - Organic compounds containing nitrogen containing nitrogen in a heterocyclic ring
C08F 8/30 - Introducing nitrogen atoms or nitrogen-containing groups
C08G 59/14 - Polycondensates modified by chemical after-treatment
C07C 209/78 - Preparation of compounds containing amino groups bound to a carbon skeleton from amines, by reactions not involving amino groups, e.g. reduction of unsaturated amines, aromatisation, or substitution of the carbon skeleton from carbonyl compounds, e.g. from formaldehyde, and amines having amino groups bound to carbon atoms of six-membered aromatic rings, with formation of methylene-diarylamines
C07C 223/00 - Compounds containing amino and —CHO groups bound to the same carbon skeleton
C07C 249/02 - Preparation of compounds containing nitrogen atoms doubly-bound to a carbon skeleton of compounds containing imino groups
73.
PROCESS FOR FERMENTIVE PREPARATION OF ALCOHOLS AND RECOVERY OF PRODUCT
This invention relates to a process for recovering an alcohol from a fermentation broth using liquid-liquid extraction, wherein at least one ionic liquid is used as the extractive solvent.
Bacteria that are not natural butanol producers were found to have increased tolerance to butanol when the membrane content of unsaturated trans fatty acids was increased. Feeding cells with unsaturated trans fatty acids increased their concentration in the membrane, which may also be accomplished by expressing a fatty acid cistrans isomerase.
The present invention relates to a process for recovering ethanol or butanol from fermentation broth using liquid-liquid extraction, wherein at least one ionic liquid is used as the extractive solvent.
Lactic acid bacterial (LAB) cells were modified such that they have a specific activity of dihydroxy-acid dehydratase enzyme activity that is increased to about 0.1 µmol min-1 mg-1. LAB cells with even higher activities of 0.2 to 0.6 µmol min-1 mg-1 of DHAD activity were obtained. These modified cells may be used to produce isobutanol when additional isobutanol biosynthetic pathway enzymes are expressed.
Yeast strains were engineered that have increased activity of heterologous proteins that require binding of an Fe-S cluster for their activity. The yeast strains have reduced activity of an endogenous Fe-S protein. Activities of heterologous fungal or plant 2Fe-2S dihydroxy-acid dehydratases and Fe-S propanediol dehydratase reactivase were increased for increased production of products made using biosynthetic pathways including these enzymes, such as valine, isoleucine, leucine, pantothenic acid (vitamin B5), isobutanol, 2-butanone and 2-butanol.
A high flux of metabolites from pyruvate to 2,3-butanediol in Lactobacillus plantarum was achieved through genetic engineering. Substantial elimination of lactate dehydrogenase activity in the presence of heterologously expressed butanediol dehydrogenase activity led to 2,3 butanediol production that was at least 49% of the total of major pyruvate-derived products.
A group of bacterial dihydroxy-acid dehydratases having a [2Fe-2S] cluster was discovered. Bacterial [2Fe-2S] DHADs were expressed as heterologous proteins in bacteria and yeast cells, providing DHAD activity for conversion of 2,3-dihydroxyisovalerate to α-ketoisovalerate or 2,3-dihydroxymethylvalerate to α-ketomethylvalerate. Isobutanol and other compounds may be synthesized in pathways that include bacterial [2Fe-2S] DHAD activity.
An E. coli host strain was engineered wherein genes adhE, ldhA, frdB, and pflB were disrupted and novel butanol dehydrogenase gene, sadB, from Achromobacter xylosoxidans, was added to produce the isobutanol production host.
A method of making butanol from at least one fermentable carbon source that overcomes the issues of toxicity resulting in an increase in the effective titer, the effective rate, and the effective yield of butanol production by fermentation utilizing a recombinant microbial host wherein the butanol is extracted into specific organic extractants during fermentation.
A process for determining the distillation characteristics of a liquid petroleum product that contains an azeotropic mixture of an oxygenated or nitrogen-containing component and at least one petroleum blending component.
Yeast cells with a reduced general control response to amino acid starvation were found to have increased tolerance to butanol in the growth medium. The reduced response was engineered by genetic modification of a gene involved in the response, a GCN gene, to eliminate activity of the encoded protein. Yeast strains with an engineered butanol biosynthetic pathway and a genetic modification in a gene involved in the general control response to amino acid starvation, which have increased butanol tolerance, are useful for production of butanol.
A method for producing a gasoline blend having a high concentration of a butanol isomer and having good cold start and warm-up driveability performance.
Methods for the fermentive production of four carbon alcohols are provided. Specifically, butanol, preferably 2-butanol is produced by the fermentive growth of a recombinant bacteria expressing a 2- butanol biosynthetic pathway. The recombinant microorganisms and methods of the invention can also be adapted to produce 2-butanone, an intermediate in the 2-butanol biosynthetic pathways disclosed herein. Specifically disclosed herein are the use of coenzyme B12-independent butanediol dehydratases that catalyzes the substrate to product conversion of 2,3-butanediol to 2-butanone in the process of producing 2-butanol and 2-butanone.
C12N 1/00 - Microorganisms, e.g. protozoaCompositions thereofProcesses of propagating, maintaining or preserving microorganisms or compositions thereofProcesses of preparing or isolating a composition containing a microorganismCulture media therefor
C12N 1/21 - BacteriaCulture media therefor modified by introduction of foreign genetic material
C12N 1/32 - Processes using, or culture media containing, lower alkanols, i.e. C1 to C6
From a bacterial strain isolated from an environmental sample, after enrichment in medium containing 1-butanol as the carbon source, a new enzyme with butanol dehydrogenase activity was identified. The enzyme can convert butyraldehyde to 1-butanol, isobutyraldehyde to isobutanol, as well as 2-butanone to 2-butanol and thus is useful for biosynthesis of butanol in recombinant microbial hosts producing these substrates. The encoding gene, named sadB, was isolated from the strain identified as an isolate of Achromobacter xylosoxidans.
Compositions of oxygenated gasolines containing isobutanol are disclosed that have reduced vapor pressure compared to those containing a single oxygenate and no isobutanol. Such compositions can be formed at a refinery or at a terminal. Methods of reducing vapor pressure of an oxygenated gasoline are disclosed and methods of reducing vapor pressure constraints upon a refinery in the production of oxygenated gasoline are disclosed. Fundamental properties of isobutanol are disclosed inctuding IR spectrum analysis. Processes and methods for blending and distributing these fuels are also disclosed.
The use of at least one C4 alcohol to increase the vapour pressure of a fuel for spark-ignition engines comprising base fuel and ethanol in which use the at least one C4 alcohol is added to a fuel comprising base fuel and ethanol to produce a resultant fuel mixture comprising 15-50 % by volume of base fuel, 15-65 % by volume of ethanol and 15 to 50 % C4 alcohol. The resultant fuel mixture exhibits an adequately high vapour pressure, for example, which meets the specification, and also exhibits a good start behaviour even at winter temperatures.
Using screening of transposon random insertion mutants, genes involved in a complex that is a three-component proton motive force-dependent multidrug efflux system were found to be involved in E. coli cell response to butanol. Reduced production of the AcrA and/or AcrB proteins of the complex confers increased butanol tolerance. E. coli strains with reduced AcrA or AcrB production and having a butanol or 2-butanone biosynthetic pathway are useful for production of butanol or 2-butanone.
C12N 15/00 - Mutation or genetic engineeringDNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purificationUse of hosts therefor
Methods for the evolution of NADPH binding ketol-acid reductoisomerase enzymes to acquire NADH binding functionality are provided. Specific mutant ketol-acid reductoisomerase enzymes isolated from Pseudomonas that have undergone co-factor switching to bind NADH are described.
Using screening of transposon random insertion mutants, genes involved in accumulation of (p)ppGpp were found to be involved in bacterial cell response to butanol. Reduced production of proteins with enzymatic activity for (p)ppGpp biosynthesis confers increased butanol tolerance. Bacterial strains with reduced (p)ppGpp accumulation and having a butanol or 2-butanone biosynthetic pathway are useful for production of butanol or 2-butanone.
Screening of fatty acid fed bacteria which are not natural butanol producers identified increased membrane cyclopropane fatty acid as providing improved butanol tolerance. Increasing expression of cyclopropane fatty acid synthase in the presence of the enzyme substrate that is either endogenous to the cell or fed to the cell, increased butanol tolerance. Bacterial strains with increased cyclopropane fatty acid synthase and having a butanol biosynthetic pathway are useful for production of butanol.
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