Abstract: The present disclosure comprises methods and compositions comprising a plant transforming bacterium of the Order Rhizobiales comprising conditional negative selectable marker genes.

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: 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: 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 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: Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing carotenoids (e.g., ?-carotene, lycopene, lutein, zeaxanthin, canthaxanthin, astaxanthin) are provided. The strains may also be engineered to co-produce at least one ?-3/?-6 polyunsaturated fatty acid and/or at least one additional antioxidant. Methods of using the carotenoid products obtained (e.g., biomass and/or pigmented oils) in food and feed applications are also provided.

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: 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 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: 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: 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: 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: 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 strains possess at least one peroxisome biogenesis factor protein knockout. Methods for producing EPA within said host cells, oils obtained from the cells, and products therefrom are claimed.

Abstract: Aquaculture feed compositions having a ratio of concentration of omega-3 polyunsaturated fatty acids of eicosapentaenoic acid [“EPA”; cis-5,8,11,14,17-eicosapentaenoic acid; 20:5] to concentration of docosahexaenoic acid [“DHA”; cis-4,7,10,13,16,19-docosahexaenoic acid; 22:6] greater than 2:1 based on the individual concentrations of EPA and DHA in the aquaculture feed composition, and having a biomass source of carotenoid(s), are disclosed. Furthermore, such aquaculture feed compositions may have a concentration of the sum of EPA and DHA that is at least about 0.8%, measured as a weight percent of the aquaculture feed composition.

Abstract: Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing carotenoids (e.g., ?-carotene, lycopene, lutein, zeaxanthin, canthaxanthin, astaxanthin) are provided. The strains may also be engineered to co-produce at least one ?-3/?-6 polyunsaturated fatty acid and/or at least one additional antioxidant. Methods of using the carotenoid products obtained (e.g., biomass and/or pigmented oils) in food and feed applications are also provided.

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: 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: 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 strains possess at least one peroxisome biogenesis factor protein knockout. Methods for producing EPA within said host cells, oils obtained from the cells, and products therefrom are claimed.

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: 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.