Butamax Advanced Biofuels LLC

E.i. Dupont De Nemours and Company

Michael Dauner

Jean-Francois Tomb

Quinn Qun Zhu

Lori Ann Maggio-Hall

Seung-Pyo Hong

Zhixiong Xue

Gopal K Chotani

Caroline M Peres

Daniel P O'Keefe

Mark J Nelson

Jahnavi Chandra Prasad

Ritu Bhalla

Dongming Xie

Hyeryoung Yoon

Robert DiCosimo

Narendra S Yadav

Douglas Robert Anton

Larry Cameron Anthony

Arthur Leo Kruckeberg

Brian James Paul

Pamela L Sharpe

Wonchul Suh

Robert A Larossa

Xiaochun Fan

Mehmedalija Jahic

Joseph Frederich Tuminello

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.

Recombinant host cells comprising phosphoketalase

Provided herein are glycerol-3-phosphate dehydrogenase (GPD) enzymes with increased Kfor NADH and GPD enzymes with substantially the same affinity for NADH and NADPH and/or are feedback inhibited by glycerol-3-phosphate. Also provided herein are recombinant microorganisms comprising a heterologous gene encoding GPD and a deletion or disruption in an endogenous gene encoding GPD. Also provided are recombinant microorganisms comprising a heterologous gene encoding GPD and a butanol biosynthetic pathway. Further provided are methods of producing butanol comprising providing the recombinant microorganisms described herein and contacting the recombinant microorganism with at least one fermentable carbon substrate under conditions wherein butanol is produced.

Glycerol 3-phosphate dehydrogenase for butanol production

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.

Competitive growth and/or production advantage for butanologen microorganism

Recombinant microbial cells are disclosed herein that produce an oil comprising at least 28 percent eicosapentaenoic acid (EPA) measured as a weight percent of dry cell weight. These cells may comprise a polynucleotide sequence encoding an active acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT) comprising at least one amino acid mutation in a membrane-bound O-acyltransferase motif. In addition, the cells may comprise a down-regulation of an endogenous polynucleotide sequence encoding Sou2 sorbitol utilization protein, and/or one or more polynucleotides encoding phospholipid:diacylglycerol acyltransferase (PDAT), delta-12 desaturase, a dihomo-gamma-linolenic acid (DGLA) synthase multizyme, delta-8 desaturase, malonyl-CoA synthetase (MCS), or acyl-CoA:lysophosphatidic acid acyltransferase (LPAAT). Also disclosed are methods of using the recombinant microbial cells to produce oil containing omega-3 polyunsaturated fatty acids such as EPA.

Recombinant microbial cells that produce at least 28% eicosapentaenoic acid as dry cell weight

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.

Recombinant host cells comprising phosphoketolases

Recombinant microbial cells are disclosed herein that comprise (i) a down-regulation of an endogenous polynucleotide sequence encoding Sou2 sorbitol utilization protein, and (ii) a polyunsaturated fatty acid (PUFA) biosynthetic pathway. The down-regulation of the polynucleotide sequence encoding Sou2 sorbitol utilization protein can increase the lipid content of the microbial cells and/or decrease the total amount of sugar alcohols produced by the microbial cells. Also disclosed are methods of using the recombinant microbial cells to produce oil containing omega-3 polyunsaturated fatty acids such as EPA.

Down-regulation of a polynucleotide encoding a Sou2 sorbitol utilization protein to modify lipid production in microbial cells

Glycerol 3- phosphate dehydrogenase for butanol production

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.

Genetic switches for butanol production

The invention relates generally to the field of industrial microbiology and alcohol production. More specifically, the invention relates to the use of thiamine, biosynthetic precursors of thiamine, nicotinic acid, nicotinamid, nicotinic acid riboside, nicotinamid riboside, or other biosynthetic precursors of nicotine adenine dinucleotide (NAD) to improve butanol production. Butanol production can be improved by providing sufficient amounts of thiamine, biosynthetic precursors of thiamine, nicotinic acid, nicotinamid, nicotinic acid riboside, nicotinamid riboside, or other biosynthetic precursors of nicotine adenine dinucleotide (NAD) in the production media.

Use of thiamine and nicotine adenine dinucleotide for butanol production

Diols produced in fermentation are processed in broth by esterification of the product diol with a carboxylic acid (e.g., fatty acid) and a catalyst (e.g., lipase) capable of esterifying the product diol, such as 1,3-propanediol, with the carboxylic acid to form the diol esters. The diol esters can be extracted from the broth, and the product diol recovered from the diol esters. The carboxylic acid can also serve as an extractant for removal of the diol esters from the fermentation medium.

Enzymatic production of alcohol esters for recovery of diols produced by fermentation

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Wilmington, DE, United States of America

Michael Dauner