E.i. Dupont De Nemours and Company

Zhixiong Xue

Quinn Qun Zhu

Narendra S Yadav

Howard Glenn Damude

Hongxiang Zhang

Daniel Joseph Macool

Seung-Pyo Hong

Dana M Walters Pollak

Stephen K Picataggio

Pamela L Sharpe

James John Ragghianti

Brian McGonigle

Peter John Gillies

Qun Zhu

Lixuan Lisa Huang

Dongming Xie

Michael Dauner

John E Seip

Hyeryoung Yoon

Hongxian He

Fateme Sima Sariaslani

Wei Wei Qi

Michael W Bostick

Ethel Noland Jackson

Todd M Vannelli

Melissa D Bosak

Yehong Jamie Wang

David Richard Short

Michael P McCluskey

Sabine Breinig

Dana M Pollak

Dana Walters M Pollak

Brian Mcgonigle

Dana Walters Pollak

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

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

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.

[object Object]

Transgenicspecies are disclosed herein that comprise a polynucleotide encoding a cytosolic malic enzyme, a lipid content that is at least about 35% by weight of the dry cell weight of thespecies, and an engineered polyunsaturated fatty acid (PUFA) biosynthetic pathway, wherein overexpression of the cytosolic malic enzyme increases lipid content.

Transgenic oleaginous yeast having increased oil content comprising increased Yap1 transcription factor activity, wherein the increased oil content is compared to the oil content of a non-transgenic oleaginous yeast, are described herein. The increased Yap1 transcription factor activity results from overexpressing a Yap1 transcription factor, by increasing the interaction between the transcription factor and a protein that is capable of activating the transcription factor, or by a combination thereof. Methods of using these yeast strains are also described.

Increased oil content by increasing Yap1 transcription factor activity in oleaginous yeasts

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding delta-8 desaturases along with a method of making long-chain polyunsaturated fatty acids (PUFAs) using these delta-8 desaturases in plants and oleaginous yeast.

Delta-8 desaturases and their use in making polyunsaturated fatty acids

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, are disclosed. 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

Engineered strains of the oleaginous yeastcapable of producing greater than 25% eicosapentaenoic acid (EPA, an &#x3c9;-3 polyunsaturated fatty acid) in the total oil fraction are described. These strains comprise various chimeric genes expressing heterologous desaturases, elongases and acyltransferases and optionally comprise various native desaturase and acyltransferase knockouts to enable synthesis and high accumulation of EPA. Production host cells are claimed, as are methods for producing EPA within said host cells.

Engineered strains of the oleaginous yeastcapable of producing greater than 50 weight percent of eicosapentaenoic acid in the total oil fraction are described. These strains over-express heterologous &#x394;9 elongases, &#x394;8 desaturases, &#x394;5 desaturases, &#x394;17 desaturases, &#x394;12 desaturases and Celongases, and optionally over-express diacylglycerol cholinephosphotransferases. Gene knockouts are also described, as are methods for producing EPA within host cells, and products comprising EPA from the optimizedstrains.

Engineered strains of the oleaginous yeastare disclosed herein that are capable of producing microbial oil comprising greater than 25 weight percent of eicosapentaenoic acid ['EPA'], an omega-3 polyunsaturated fatty acid, measured as a weight percent of dry cell weight.

Recombinant microbial host cells for high eicosapentaenoic acid production

Acyltransferases are provided, suitable for use in the manufacture of microbial oils enriched in omega fatty acids in oleaginous yeast (e.g.,). Specifically, genes encoding diacylglycerol acyltransferase (DGAT1) have been isolated fromand. These genes encode enzymes that participate in the terminal step in oil biosynthesis in yeast. Each is expected to play a key role in altering the quantity of polyunsaturated fatty acids produced in oils of oleaginous yeasts.

Diacylglycerol acyltransferases for alteration of polyunsaturated fatty acids and oil content in oleaginous organisms

An engineered strain of the oleaginous yeastcapable of producing greater than 5.6% docosahexaenoic acid acid (DHA, an w-3 polyunsaturated fatty acid) in the total oil fraction is described. This strain comprises various chimeric genes expressing heterologous desaturases, elongases and acyltransferases and optionally comprises various native desaturase and acyltransferase knockouts to enable synthesis and high accumulation of DHA. Production host cells are claimed, as are methods for producing DHA within said host cells.

Growing community of inventors

Chadds Ford, PA, United States of America

Zhixiong Xue