# US 8,785,163 B2 · Desaturases and methods for producing polyunsaturated fatty acids in transgenic organisms > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,163 B2 - Document ID: US8785163B2 - Status: Granted - Kind: B2 - Application number: 12/671,090 - Filed: 2008-07-30 - Published: 2010-08-05 - Granted: 2014-07-22 - Earliest claimed: 2007-07-31 - Assignees: BASF Plant Science GmbH - Applicants: Toralf Senger; Jörg Bauer - Inventors: Toralf Senger; Jörg Bauer - Web page: https://patentgrep.com/patent/US8785163B2 ## CPC - C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition} - C12N 9/0083: {Miscellaneous (1.14.99)} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to polynucleotides from Helobdella robusta, Laccaria bicolor, Lottia gigantea, Microcoleus chthonoplastes, Monosiga brevicollis, Mycosphaerella fijiensis, Mycospaerella graminicola, Naegleria gruberi, Nectria haematococca, Nematostella vectensis, Phycomyces blakesleeanus, Trichoderma resii, Physcomitrella patens, Postia placenta, Selaginella moellendorffii and Microdochium nivale, which code for desaturases and which can be employed for the recombinant production of polyunsaturated fatty acids. The invention furthermore relates to vectors, host cells and transgenic nonhuman organisms which comprise the polynucleotides according to the invention, and to the polypeptides encoded by the polynucleotides. The invention furthermore relates to antibodies against the polypeptides according to the invention. Finally, the invention also relates to production processes for the polyunsaturated fatty acids and for oil, lipid and fatty acid compositions and to their use as drugs, cosmetics, foodstuffs, feedstuffs, preferably fish food, or food supplements. ## Claims ### Claim 1 An isolated polynucleotide comprising a nucleic acid sequence operably linked to a heterologous expression control sequence, wherein said nucleic acid sequence encodes a polypeptide having delta-15 desaturase activity and comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 133, and wherein said amino acid sequence further comprises the amino acid motif of SEQ ID NO: 116 and any of the amino acid motifs of SEQ ID NO: 83, SEQ ID NO: 84 and SEQ ID NO: 85. ### Claim 2 (depends on 1) The isolated polynucleotide of claim 1, wherein the polynucleotide consists of RNA or DNA. ### Claim 3 (depends on 1) A vector comprising the isolated polynucleotide of claim 1. ### Claim 4 (depends on 3) The vector according to claim 3, wherein the vector is an expression vector. ### Claim 5 (depends on 3) The vector according to claim 3, wherein the vector comprises at least one further polynucleotide which codes for a further enzyme which is involved in the biosynthesis of lipids or fatty acids. ### Claim 6 (depends on 5) The vector of claim 5, wherein the enzyme is selected from the group consisting of: acyl-CoA dehydrogenase(s), acyl-ACP [=acyl carrier protein] desaturase(s), acyl-ACP thioesterase(s), fatty acid acyltransferase(s), acyl-CoA:lysophospholipid acyltransferase(s), fatty acid synthase(s), fatty acid hydroxylase(s), acetyl-coenzyme A carboxylase(s), acyl-coenzyme A oxidase(s), fatty acid desaturase(s), fatty acid acetylenase(s), lipoxygenase(s), triacylglycerol lipase(s), allene oxide synthase(s), hydroperoxide lyase(s), fatty acid elongase(s), Δ4-desaturase(s), Δ5-desaturase(s), Δ6-desaturase(s), Δ8-desaturase(s), Δ9-desaturase(s), Δ12-desaturase(s), Δ5-elongase(s), Δ6-elongase(s), and Δ9-elongase(s). ### Claim 7 (depends on 1) A host cell comprising: (a) the isolated polynucleotide of claim 1, or (b) a vector comprising said isolated polynucleotide. ### Claim 8 (depends on 7) The host cell according to claim 7, wherein the host cell additionally comprises at least one further enzyme which is involved in the biosynthesis of lipids or fatty acids. ### Claim 9 (depends on 3) A host cell comprising the vector of claim 3. ### Claim 10 (depends on 8) The host cell of claim 8, wherein the enzyme is selected from the group consisting of: acyl-CoA dehydrogenase(s), acyl-ACP [=acyl carrier protein] desaturase(s), acyl-ACP thioesterase(s), fatty acid acyltransferase(s), acyl-CoA:lysophospholipid acyltransferase(s), fatty acid synthase(s), fatty acid hydroxylase(s), acetyl-coenzyme A carboxylase(s), acyl-coenzyme A oxidase(s), fatty acid desaturase(s), fatty acid acetylenase(s), lipoxygenase(s), triacylglycerol lipase(s), allene oxide synthase(s), hydroperoxide lyase(s), fatty acid elongase(s), Δ4-desaturase(s), Δ5-desaturase(s), Δ6-desaturase(s), Δ8-desaturase(s), Δ9-desaturase(s), Δ12-desaturase(s), Δ5-elongase(s), Δ6-elongase(s) and Δ9-elongase(s). ### Claim 11 (depends on 1) A method of generating a polypeptide with delta-15 desaturase activity, comprising: (a) expressing the isolated polynucleotide of claim 1 in a host cell; and (b) obtaining, from the host cell, the polypeptide which is encoded by the polynucleotide according to (a). ### Claim 12 (depends on 1) An isolated polypeptide which is encoded by the polynucleotide according to claim 1. ### Claim 13 (depends on 1) A transgenic, nonhuman organism comprising the polynucleotide of claim 1. ### Claim 14 (depends on 13) The transgenic, nonhuman organism according to claim 13, wherein the organism is an animal, a plant or a multicellular microorganism. ### Claim 15 (depends on 7) A process for the production of a substance which has the structure shown in the general formula I where R1=hydroxyl, coenzyme A (thioester), lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysodiphosphatidylglycerol, lysophosphatidylserine, lysophosphatidylinositol, sphingo base or a radical of the formula II R2=hydrogen, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysodiphosphatidylglycerol, lysophosphatidylserine, lysophosphatidylinositol or saturated or unsaturated C2-C24-alkylcarbonyl, R3=hydrogen, saturated or unsaturated C2-C24-alkylcarbonyl, or R2 and R3 independently of one another are a radical of the formula Ia: n=2, 3, 4, 5, 6, 7 or 9, m=2, 3, 4, 5 or 6; and p=0 or 3; and wherein the process comprises cultivating the host cell of claim 7 under conditions which permit the biosynthesis of the substance. ### Claim 16 (depends on 15) A process for the production of an oil, lipid or fatty acid composition, comprising the steps of the process according to claim 15 and the further step of formulating the substance as an oil, lipid or fatty acid composition. ### Claim 17 (depends on 16) The process according to claim 16, wherein the oil, lipid or fatty acid composition is formulated further to give a drug, a cosmetic product, a foodstuff, a feedstuff, a fish food, or a food supplement. ### Claim 18 (depends on 13) A process for the production of a substance which has the structure shown in the general formula I where R1=hydroxyl, coenzyme A (thioester), lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysodiphosphatidylglycerol, lysophosphatidylserine, lysophosphatidylinositol, sphingo base or a radical of the formula II R2=hydrogen, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysodiphosphatidylglycerol, lysophosphatidylserine, lysophosphatidylinositol or saturated or unsaturated C2-C24-alkylcarbonyl, R3=hydrogen, saturated or unsaturated C2-C24-alkylcarbonyl, or R2 and R3 independently of one another are a radical of the formula Ia: n=2, 3, 4, 5, 6, 7 or 9, m=2, 3, 4, 5 or 6; and p=0 or 3; and wherein the process comprises cultivating the transgenic, non-human organism of claim 13 under conditions which permit the biosynthesis of the substance. ### Claim 19 (depends on 18) A process for the production of an oil, lipid or fatty acid composition, comprising the steps of the process according to claim 18 and the further step of formulating the substance as an oil, lipid or fatty acid composition. ### Claim 20 (depends on 19) The process according to claim 19, wherein the oil, lipid or fatty acid composition is formulated further to give a drug, a cosmetic product, a foodstuff, a feedstuff, a fish food, or a food supplement. ### Claim 21 (depends on 1) The isolated polynucleotide of claim 1, wherein the nucleic acid sequence is selected from the group consisting of: (a) the nucleic acid sequence of SEQ ID NO: 132; (b) a nucleic acid sequence which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 133; and (c) a nucleic acid sequence which encodes a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 133 and having delta-15 desaturase activity. ### Claim 22 (depends on 21) The isolated polynucleotide of claim 21, wherein the nucleic acid sequence encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 133 and having delta-15 desaturase activity. ### Claim 23 (depends on 1) The isolated polynucleotide of claim 1, wherein the nucleic acid sequence comprises: (a) the nucleic acid sequence of SEQ ID NO: 132; or (b) a nucleic acid sequence which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 133. ### Claim 24 (depends on 1) The isolated polynucleotide of claim 1, wherein the amino acid motif of SEQ ID NO: 116 is the amino acid motif of SEQ ID NO: 117, SEQ ID NO: 118, or SEQ ID NO: 119.