E.i. Dupont De Nemours and Company

Danisco US Inc.

Dupont Industrial Biosciences Usa, LLC

Corteva Agriscience LLC

Quinn Qun Zhu

Zhixiong Xue

Howard Glenn Damude

Narendra S Yadav

Stephen K Picataggio

Seung-Pyo Hong

Hongxiang Zhang

Dana M Walters Pollak

Pamela L Sharpe

Daniel Joseph Macool

James John Ragghianti

Brian McGonigle

Hongxian He

Peter John Gillies

John E Seip

Michael W Bostick

Dongming Xie

Der-Ing Liao

Michael Dauner

Anthony John Kinney

Xiaochun Fan

Hyeryoung Yoon

Brian Mcgonigle

Rebecca E Cahoon

Saverio Carl Falco

J Antoni Rafalski

Zhongsen Li

Hajime Sakai

Edgar Benjamin Cahoon

Rick Weizhang Ye

Guo-Hua Miao

Yougen Li

Yiwen Fang

Peizhong Zheng

Kevin G Ripp

Joan Tellefsen Odell

Lixuan Lisa Huang

Jian-Ming Lee

Sabine Susanne Hantke

Michele Morgante

Xiping Niu

Paula Johanna Maria Teunissen

Min Qi

Ethel Noland Jackson

Melissa D Bosak

Yehong Jamie Wang

David Richard Short

Despina Boughioukou

Himanshu H Dhamankar

Dana Walters Pollak

Dana M Pollak

Described are compositions and methods relating to modified yeast that over-express polyadenylate-binding protein (PAB1). The yeast produces a deceased amount of acetate compared to parental cells. Such yeast is particularly useful for large-scale ethanol production from starch substrates where acetate in an undesirable end product.

Reduction in acetate production by yeast over-expressing PAB1

Recombinant microbial cells comprising an engineered LCDA production pathway that comprises at least one up-regulated long-chain acyl-CoA synthetase (ACoS) are disclosed. These recombinant microbial cells are capable of producing one or more long-chain dicarboxylic acid (LCDA) products from a long-chain fatty acid-comprising substrate. Methods of using recombinant microbial cells to produce LCDAs are also disclosed.

High level production of long-chain dicarboxylic acids with microbes

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

Methods of increasing the amount of polyunsaturated fatty acids (PUFAs) in the total lipid fraction and in the oil fraction of PUFA-producing, oleaginous eukaryotes, accomplished by modifying the activity of peroxisome biogenesis factor (Pex) proteins. Disruptions of a chromosomal Pex3 gene, Pex10p gene or Pex16p gene in a PUFA-producing, oleaginous eukaryotic strain resulted in an increased amount of PUFAs, as a percent of total fatty acids and as a percent of dry cell weight, in the total lipid fraction and in the oil fraction of the strain, as compared to the parental strain whose native Pex protein was not disrupted.

Peroxisome biogenesis factor protein (Pex) disruptions for altering polyunsaturated fatty acids and total lipid content in oleaginous eukaryotic organisms

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

Acyl-CoA:lysophosphatidylcholine acyltransferase ['LPCAT'] having the ability to convert acyl-CoA+1-acyl-sn-glycero-3-phosphocholine to CoA+1,2-diacyl-sn-glycero-3-phosphocholine (EC 2.3.1.23) is disclosed herein to be over-expressed along with the over-expression of phospholipid:diacylglycerol acyltransferase ['PDAT'] having the ability to transfer a fatty acyl group from the sn-2 position of a phospholipid (e.g., phosphatidylcholine) to the sn-3 position of 1,2-diacylglycerol [E.C.2.3.1.158], thus resulting in a lysophospholipid and TAG. Co-expression of these enzymes in a recombinant microbial host cell resulted in increased production of long chain polyunsaturated fatty acids [&#x201c;PUFAs&#x201d;].

Production of polyunsaturated fatty acids by coexpression of acyl-CoA:lysophosphatidylcholine acyltransferases and phospholipid:diacylglycerol acyltransferases

Mutant acyl-CoA:lysophosphatidylcholine acyltransferases ['LPCATs'] having the ability to convert acyl-CoA+1-acyl-sn-glycero-3-phosphocholine to CoA+1,2-diacyl-sn-glycero-3-phosphocholine (EC 2.3.1.23) are disclosed herein. Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding mutant LPCATs, along with a method of making long chain polyunsaturated fatty acids ['PUFAs'] using these mutant LPCATs in oleaginous yeast, are also disclosed.

Nucleic acids encoding mutant acyl-CoA:lysophosphatidylcholine acyltransferases

Described are methods of reducing the amount of byproduct organic acids during fermentation of an organism, based on expression of a heterologous malonyl-CoA synthetase. A polyunsaturated fatty acid ['PUFA']-producing strain of the oleaginous yeastwas engineered to express a heterologous malonyl-CoA synthetase gene. The expression did not effect the production of PUFAs, but did result in a reduced amount of malonates when compared to the amount of malonates produced in the parental strain not expressing malonyl-CoA synthetase.

Reducing byproduction of malonates by yeast in a fermentation process

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding delta-9 elongases along with a method of making long-chain polyunsaturated fatty acids (PUFAs) using these delta-9 elongases in plants.

Delta-9 elongases and their use in making polyunsaturated fatty acids

The present invention relates to methods for the production of &#x3c9;-3 and/or &#x3c9;-6 fatty acids in oleaginous yeast. Thus, desaturases and elongases able to catalyze the conversion of linoleic acid (LA) to &#x3b3;-linolenic acid (GLA); &#x3b1;-linoleic acid (ALA) to stearidonic acid (STA); GLA to dihomo-&#x3b3;-linoleic acid (DGLA); STA to eicosatetraenoic acid (ETA); DGLA to arachidonic acid (ARA); ETA to eicosapentaenoic acid (EPA); DGLA to ETA; EPA to docosapentaenoic acid (DPA); and ARA to EPA have been introduced into the genome offor synthesis of ARA and EPA.

Production of polyunsaturated fatty acids in oleaginous yeasts

Methods for increasing Cto Celongation conversion efficiency and/or &#x394;4 desaturation conversion efficiency in long-chain polyunsaturated fatty acid ['LC-PUFA']-producing recombinant oleaginous microbial host cells are provided herein, based on over-expression of acyl-CoA:lysophospholipid acyltransferases ['LPLATs'] (e.g., Ale1, LPAAT, LPCAT). Production host cells and oils produced by the methods of the invention are also claimed.

Long chain omega-3 and omega-6 polyunsaturated fatty acid biosynthesis by expression of acyl-CoA lysophospholipid acyltransferases

Mutant delta-9 elongases having the ability to convert linoleic acid [18:2, LA] to eicosadienoic acid [20:2, EDA] and/or &#x3b1;-linolenic [18:3, ALA] to eicosatrienoic acid [20:3, ETrA] are disclosed herein. Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding mutant delta-9 elongases, along with a method of making long chain polyunsaturated fatty acids ['PUFAs'] using these mutant delta-9 elongases in oleaginous yeast are also disclosed.

Mutant delta-9 elongases and their use in making polyunsaturated fatty acids

Promoter regions associated with thediacylglycerol acyltransferase 2 (dgat2) gene are disclosed and have been found to be particularly effective for the expression of heterologous genes in yeast. These promoter regions will be useful for driving high-level expression of genes involved in the production of omega-3 and omega-6 fatty acids.

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Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding delta-5 desaturase along with a method of making long chain polyunsaturated fatty acids (PUFAs) using this delta-5 desaturase in plants and oleaginous yeast are disclosed.

Delta-5 desaturase and its use in making polyunsaturated fatty acids

Growing community of inventors

West Chester, PA, United States of America

Quinn Qun Zhu