# US 7,608,753 B2 · PUFA polyketide synthase systems and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,608,753 B2 - Document ID: US7608753B2 - Status: Granted - Kind: B2 - Application number: 11/778,575 - Filed: 2007-07-16 - Published: 2008-02-21 - Granted: 2009-10-27 - Earliest claimed: 1999-01-14 - Assignees: Martek Biosciences Corporation - Inventors: James G. Metz; Craig A. Weaver; William R. Barclay; James H. Flatt - Web page: https://patentgrep.com/patent/US7608753B2 ## CPC - C12N 15/52: Genes encoding for enzymes or proenzymes - A23D 7/001: {Spread compositions (characterised by ingredients other than fatty acid triglycerides A23D7/0056)} - A23L 33/115: Fatty acids or derivatives thereof; Fats or oils - A23L 33/135: Bacteria or derivatives thereof, e.g. probiotics - C07K 14/28: from Vibrionaceae (F) - C12P 7/6434: Docosahexenoic acids [DHA] - C12P 7/6472: containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A23D: EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS (preservation thereof A23B20/00; production, refinement or preservation of animal or vegetable fats or oils C11B, C11C) - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} ## Abstract The invention generally relates to polyunsaturated fatty acid (PUFA) polyketide synthase (PKS) systems, to homologues thereof, to isolated nucleic acid molecules and recombinant nucleic acid molecules encoding biologically active domains of such a PUFA PKS system, to genetically modified organisms comprising PUFA PKS systems, to methods of making and using such systems for the production of bioactive molecules of interest, and to novel methods for identifying new bacterial and non-bacterial microorganisms having such a PUFA PKS system. ## Claims ### Claim 1 A genetically modified plant, wherein cells of the plant are transfected with at least one recombinant nucleic acid molecule encoding a polyketide synthase (PKS) system for the production of polyunsaturated fatty acids (PUFAs), wherein the PKS system comprises the following proteins: a) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:39 and has β-keto acyl-ACP synthase (KS) activity, malonyl-CoA:ACP acyltransferase (MAT) activity, acyl carrier protein (ACP) activity and ketoreductase (KR) activity; b) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:52 and has β-keto acyl-ACP synthase (KS) activity, chain length factor (CLF) activity, acyl transferase (AT) activity, and enoyl ACP-reductase (ER) activity; and c) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:62 and has FabA-like β-hydroxy acyl-ACP dehydrase (DH) activity and enoyl ACP-reductase (ER) activity. ### Claim 2 (depends on 1) The genetically modified plant of claim 1, wherein the protein of (a) comprises the amino acid sequence of SEQ ID NO:39. ### Claim 3 (depends on 1) The genetically modified plant of claim 1, wherein the protein of (b) comprises the amino acid sequence of SEQ ID NO:52. ### Claim 4 (depends on 1) The genetically modified plant of claim 1, wherein the protein of (c) comprises the amino acid sequence of SEQ ID NO:62. ### Claim 5 (depends on 1) The genetically modified plant of claim 1, wherein the PKS system comprises: a) a protein comprising the amino acid sequence of SEQ ID NO:39; b) a protein comprising the amino acid sequence of SEQ ID NO:52; and c) a protein comprising the amino acid sequence of SEQ ID NO:62. ### Claim 6 (depends on 1) The genetically modified plant of claim 1, wherein the protein of (a) is encoded by SEQ ID NO:38. ### Claim 7 (depends on 1) The genetically modified plant of claim 1, wherein the protein of (b) is encoded by SEQ ID NO:51. ### Claim 8 (depends on 1) The genetically modified plant of claim 1, wherein the protein of (c) is encoded by SEQ ID NO:61. ### Claim 9 (depends on 1) The genetically modified plant of claim 1, wherein cells of the plant have been further transfected with a phosphopantetheinyl transferase (PPTase). ### Claim 10 (depends on 1) The genetically modified plant of claim 1, wherein the plant produces docosahexaenoic acid (DHA). ### Claim 11 (depends on 1) The genetically modified plant of claim 1, wherein the plant produces docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA). ### Claim 12 (depends on 1) The genetically modified plant of claim 1, wherein the plant produces a polyunsaturated fatty acid (PUFA) profile that differs from the naturally occurring plant without a genetic modification. ### Claim 13 A recombinant plant cell, wherein the plant cell expresses at least one recombinant nucleic acid molecule encoding a polyketide synthase (PKS) system for the production of polyunsaturated fatty acids (PUFAs), wherein the PKS system comprises the following proteins: a) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:39 and has β-keto acyl-ACP synthase (KS) activity, malonyl-CoA:ACP acyltransferase (MAT) activity, acyl carrier protein (ACP) activity and ketoreductase (KR) activity; b) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:52 and has β-keto acyl-ACP synthase (KS) activity, chain length factor (CLF) activity, acyl transferase (AT) activity, and enoyl ACP-reductase (ER) activity; and c) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:62 and has FabA-like β-hydroxy acyl-ACP dehydrase (DH) activity and enoyl ACP-reductase (ER) activity. ### Claim 14 (depends on 13) The recombinant plant cell of claim 13, wherein the protein of (a) comprises the amino acid sequence of SEQ ID NO:39. ### Claim 15 (depends on 13) The recombinant plant cell of claim 13, wherein the protein of (b) comprises the amino acid sequence of SEQ ID NO:52. ### Claim 16 (depends on 13) The recombinant plant cell of claim 13, wherein the protein of (c) comprises the amino acid sequence of SEQ ID NO:62. ### Claim 17 (depends on 13) The recombinant plant cell of claim 13, wherein the PKS system comprises: a) a protein comprising the amino acid sequence of SEQ ID NO:39; b) a protein comprising the amino acid sequence of SEQ ID NO:52; and c) a protein comprising the amino acid sequence of SEQ ID NO:62. ### Claim 18 (depends on 13) The recombinant plant cell of claim 13, wherein the protein of (a is encoded by SEQ ID NO:38. ### Claim 19 (depends on 13) The recombinant plant cell of claim 13, wherein the protein of (b) is encodec by SEQ ID NO:51. ### Claim 20 (depends on 13) The recombinant plant cell of claim 13, wherein the protein of (c) is encodec by SEQ ID NO:61. ### Claim 21 A method to produce a genetically modified plant that has a polyunsaturated fatty acid (PUFA) profile that differs from the naturally occurring plant and that produces at least one long chain polyunsaturated fatty acid (PUFA), comprising transfecting plant cells with at least one recombinant nucleic acid molecule encoding a polyketide synthase (PKS) system for the production of polyunsaturated fatty acids (PUFAs) and producing the plant using the plant cells, wherein the PKS system comprises the following proteins: a) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO39 and has 13-keto acyl-ACP synthase (KS) activity, malonyl-CoA:ACP acyltransferase (MAT) activity, acyl carrier protein (ACP) activity and ketoreductase (KR) activity; b) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:52 and has 13-keto acyl-ACP synthase (KS) activity, chain length factor (CLF) activity, acyl transferase (AT) activity, and enoyl ACP-reductase (ER) activity; and c) a protein comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:62 and has FabA-like β-hydroxy acyl-ACP dehydrase (DH) activity and enoyl ACP-reductase (ER) activity.