Abstract: 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.

Abstract: Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing greater than 25% eicosapentaenoic acid (EPA, an ?-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.

Abstract: 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.

Abstract: Recombinant oleaginous microorganisms having increased oil content due to the expression of a caleosin polypeptide are described. A recombinant oleaginous microorganism of the disclosed invention produces at least 25% of its dry cell weight as oil, and comprises a functional polyunsaturated fatty acid (PUFA) biosynthetic pathway and at least one genetic construct encoding a caleosin polypeptide. A method for increasing the amount of oil in a recombinant oleaginous microorganism is also described.

Abstract: 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.

Abstract: Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing greater than 50 weight percent of eicosapentaenoic acid in the total oil fraction are described. These strains over-express heterologous ?9 elongases, ?8 desaturases, ?5 desaturases, ?17 desaturases, ?12 desaturases and C16/18 elongases, 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 optimized Yarrowia lipolytica strains.

Abstract: Disclosed herein are transformed Yarrowia lipolytica comprising an exogenous polynucleotide encoding a polypeptide having sucrose invertase activity. Also disclosed are methods of using the transformed Y. lipolytica.

Abstract: Promoter regions associated with the Yarrowia lipolytica n-alkane-hydroxylating cytochrome P450 (ALK2) 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.

Abstract: The invention provides isolated pyruvate dehydrogenase kinase nucleic acids and their encoded polypeptides. The present invention provides methods and compositions relating to altering pyruvate dehydrogenase kinase levels in plants. The invention further provides recombinant expression cassettes, host cells, transgenic plants, and antibody compositions.

Abstract: Engineered strains of the oleaginous yeast Yarrowia lipolytica are 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.

Abstract: An engineered strain of the oleaginous yeast Yarrowia lipolytica capable 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.

Abstract: Mutant delta-5 desaturases, having the ability to convert dihomo-gamma-linolenic acid [DGLA; 20:3 omega-6] to arachidonic acid [ARA; 20:4 omega-6] and/or eicosatetraenoic acid [ETA; 20:4 omega-3] to eicosapentaenoic acid [EPA; 20:5 omega-3] and possessing at least one mutation within the HPGG (SEQ ID NO:7) motif of the cytochome b5-like domain and at least one mutation within the HDASH (SEQ ID NO:8) motif are disclosed. Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding delta-5 desaturases, along with a method of making long chain polyunsaturated fatty acids [“PUFAs”], are also disclosed.

Abstract: Methods for increasing C18 to C20 elongation conversion efficiency and/or ?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.

Abstract: Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing greater than 25% eicosapentaenoic acid (EPA, an ?-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.

Abstract: Described are engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing an oil comprising greater than 50 weight percent of eicosapentaenoic acid [“EPA”], an ?-3 polyunsaturated fatty acid, measured as a weight percent of total fatty acids [“% TFAs”] and having a ratio of at least 3.1 of EPA % TFAs, to linoleic acid, measured as % TFAs. These strains over-express at least one ?9 elongase/?8 desaturase multizyme, in addition to other heterologous ?9 elongases, ?8 desaturases, ?5 desaturases, ?17 desaturases, ?12 desaturases, C16/18 elongases, and optionally over-express diacylglycerol cholinephosphotransferases, malonyl CoA synthetases and/or acyl-CoA lysophospholipid acyltransferases. The expression of at least one peroxisome biogenesis factor protein is down-regulated. Methods for producing EPA within said host cells, oils obtained from the cells, and products therefrom are claimed.

Abstract: The present invention relates to mutant ?5 desaturases, which have the ability to convert dihomo-?-linolenic acid [DGLA; 20:3 ?-6] to arachidonic acid [ARA; 20:4 ?-6] and/or eicosatetraenoic acid [ETA; 20:4 ?-3] to eicosapentaenoic acid [EPA; 20:5 ?-3] and which possess at least one mutation within the HPGG motif of the cytochome b5-like domain. Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding ?5 desaturases, along with a method of making long chain polyunsaturated fatty acids [“PUFAs”] using these mutant ?5 desaturases in oleaginous yeast, are disclosed.

Abstract: 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 [“PUFAs”].

Abstract: 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.

Abstract: Promoter regions associated with the Yarrowia lipolytica peroxisomal 2,4-dienoyl-CoA reductase (SPS19) 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.

Abstract: Transgenic Yarrowia species 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 the Yarrowia species, and an engineered polyunsaturated fatty acid (PUFA) biosynthetic pathway, wherein overexpression of the cytosolic malic enzyme increases lipid content.