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: 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: 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 yeast Yarrowia lipolytica was 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.

Abstract: 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.

Abstract: The present invention relates to methods for the production of ?-3 and/or ?-6 fatty acids in oleaginous yeast. Thus, desaturases and elongases able to catalyze the conversion of linoleic acid (LA) to ?-linolenic acid (GLA); ?-linoleic acid (ALA) to stearidonic acid (STA); GLA to dihomo-?-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 of Yarrowia for synthesis of ARA and EPA.

Abstract: 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.

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: 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) using these delta-5 desaturases in plants are disclosed.

Abstract: A recombinant microbial host cell is provided capable of producing astaxanthin from ?-carotene without a measurable concomitant accumulation of ketolated or hydroxylated intermediates such as adonixanthin, zeaxanthin, adonirubin, echinenone, 3-hydroxyechinenone, 3?-hydroxyechinenone, canthaxanthin, and ?-cryptoxanthin. Specifically, a ?-carotene producing microbial host cell was engineered to express two heterologous genes, a ?-carotene ketolase from Chlamydomonas reinhardtii in combination with a carotenoid hydroxylase from Brevundimonas vesicularis or Arabidopsis thaliana.

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: 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: Mutant delta-9 elongases having the ability to convert linoleic acid [18:2, LA] to eicosadienoic acid [20:2, EDA] and/or ?-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.

Abstract: Promoter regions associated with the Yarrowia lipolytica diacylglycerol 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.

Abstract: 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.

Abstract: 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) using these delta-5 desaturases in plants are disclosed.

Abstract: Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding multizymes (i.e., single polypeptides having at least two independent and separable enzymatic activities) along with a method of making long-chain polyunsaturated fatty acids (PUFAs) using these multizymes in plants and oleaginous yeast are disclosed.

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. 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: 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.

Abstract: Promoter regions associated with the Yarrowia lipolytica esterase/lipase (EL1) 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.