PLANT SEED OILS CONTAINING POLYUNSATURATED FATTY ACIDS
Disclosed are plants that have been genetically modified to express a PKS-like system for the production of PUFAs (a PUFA PKS system), wherein oils produced by the plant contain at least one PUFA produced by the PUFA PKS system and are free of the mixed shorter-chain and less unsaturated PUFAs that are fatty acid products produced by the modification of products of the FAS system in standard fatty acid pathways. Also disclosed are the oil seeds, oils, and products comprising such oils produced by this system, as well as methods for producing such plants.
1 . A plant or a part of the plant, wherein the total fatty acid profile in the plant or part of the plant comprises at least about 0.5% by weight of at least one polyunsaturated fatty acid (PUFA) having at least twenty carbons and four or more carbon-carbon double bonds, and wherein the total fatty acid profile in the plant or part of the plant contains less than 5% in total of all of the following PUFAs: gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds.
2 . A plant or a part of the plant, wherein the total fatty acid profile in the plant or part of the plant comprises at least about 0.5% by weight of at least one polyunsaturated fatty acid (PUFA) having at least twenty carbons and four or more carbon-carbon double bonds, and wherein the total fatty acid profile in the plant or part of the plant contains less than 1% of each of the following PUFAs: gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds.
3 . A plant or a part of the plant, wherein the total fatty acid profile in the plant or part of the plant comprises at least about 0.5% by weight of at least one polyunsaturated fatty acid (PUFA) having at least twenty carbons and four or more carbon-carbon double bonds, and wherein the total fatty acid profile in the plant or part of the plant contains less than 2% of gamma-linolenic acid (GLA; 18:3, n-6) and dihomo-gamma-linolenic acid (DGLA or HGLA; 20:3, n-6).
4 . The plant or part of the plant of claim 4 , wherein the total fatty acid profile in the plant or part of the plant contains less than 1% by weight of gamma-linolenic acid (GLA; 18:3, n-6) and dihomo-gamma-linolenic acid (DGLA or HGLA; 20:3, n-6).
5 . A plant or a part of the plant, wherein the total fatty acid profile in the plant or part of the plant comprises at least about 0.5% by weight of at least one polyunsaturated fatty acid (PUFA) having at least twenty carbons and four or more carbon-carbon double bonds, and wherein the total fatty acid profile in the plant or part of the plant contains less than 1% of gamma-linolenic acid (GLA; 18:3, n-6).
6 . The plant or part of the plant of claim 5 , wherein the total fatty acid profile in the plant or part of the plant contains less than 0.5% by weight of gamma-linolenic acid (GLA; 18:3, n-6).
7 . A plant or part of a plant, wherein the plant has been genetically modified to express enzymes that produce at least one polyunsaturated fatty acid (PUFA) having at least twenty carbons and four or more carbon-carbon double bonds, wherein the total fatty acid profile in the plant or part of the plant comprises at least about 0.5% by weight of said at least one PUFA, and wherein the total fatty acids produced by said enzymes, other than said at least one PUFA, comprise less than about 10% of the total fatty acids produced by said plant.
8 . The plant or part of the plant of claim 7 , wherein the total fatty acids produced by said enzymes, other than said at least one PUFA, comprise less than 5% by weight of the total fatty acids produced by said plant.
9 . The plant or part of the plant of claim 7 , wherein the fatty acids consisting of gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds, comprise less than 5% by weight of the total fatty acids produced by said plant
10 . The plant or part of the plant of claim 7 , wherein gamma-linolenic acid (GLA; 18:3, n-6) comprises less than 1% by weight of the total fatty acids produced by said plant.
11 . The plant or part of a plant of claim 1 , wherein the plant has not been genetically modified to express a desaturase or an elongase enzyme.
12 . A plant or part of a plant, wherein the plant has been genetically modified with a PUFA PKS system from a eukaryote that produces at least one polyunsaturated fatty acid (PUFA), and wherein the total fatty acid profile in the plant or part of the plant comprises a detectable amount of said at least one PUFA.
13 . The plant or part of a plant of claim 12 , wherein the total fatty acid profile in the plant or part of the plant comprises at least 0.5% by weight of said at least one PUFA.
14 . The plant or part of a plant of claim 12 , wherein the total fatty acids produced by said PUFA PKS system, other than said at least one PUFA, comprises less than about 10% by weight of the total fatty acids produced by said plant.
15 . The plant or part of a plant of claim 12 , wherein the total fatty acids produced by said enzymes, other than said at least one PUFA, comprises less than about 5% by weight of the total fatty acids produced by said plant.
16 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system comprises:
a) at least one enoyl-ACP reductase (ER) domain;
b) at least four acyl carrier protein (ACP) domains;
c) at least two β-ketoacyl-ACP synthase (KS) domains;
d) at least one acyltransferase (AT) domain;
e) at least one β-ketoacyl-ACP reductase (KR) domain;
f) at least two FabA-like β-hydroxyacyl-ACP dehydrase (DH) domains; and
g) at least one chain length factor (CLF) domain;
h) at least one malonyl-CoA:ACP acyltransferase (MAT) domain.
17 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system comprises:
a) two enoyl ACP-reductase (ER) domains;
b) eight or nine acyl carrier protein (ACP) domains;
c) two β-keto acyl-ACP synthase (KS) domains;
d) one acyltransferase (AT) domain;
e) one ketoreductase (KR) domain;
f) two FabA-like β-hydroxy acyl-ACP dehydrase (DH) domains;
g) one chain length factor (CLF) domain; and
h) one malonyl-CoA:ACP acyltransferase (MAT) domain.
18 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system is from a Thraustochytriales microorganism.
19 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system is from Schizochytrium.
20 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system is from Thraustochytrium.
21 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system is from a microorganism selected from the group consisting of: Schizochytrium sp. American Type Culture Collection (ATCC) No. 20888 ; Thraustochytrium 23B ATCC No. 20892, and a mutant of any of said microorganisms.
22 . The plant or part of a plant of claim 12 , wherein the nucleic acid sequences encoding the PUFA PKS system hybridize under stringent hybridization conditions to the genes encoding the PUFA PKS system from a microorganism selected from the group consisting of: Schizochytrium sp. American Type Culture Collection (ATCC) No. 20888 ; Thraustochytrium 23B ATCC No. 20892; and a mutant of any of said microorganisms.
23 . The plant or part of a plant of claim 12 , wherein the nucleic acid sequences encoding the PUFA PKS system hybridize under stringent hybridization conditions to the genes encoding the PUFA PKS system from Schizochytrium sp. American Type Culture Collection (ATCC) No. 20888 or a mutant thereof.
24 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system comprises at least one domain from a PUFA PKS system from a Thraustochytriales microorganism.
25 . A plant or part of a plant, wherein the plant has been genetically modified with a PUFA PKS system that produces at least one polyunsaturated fatty acid (PUFA), and wherein the total fatty acid profile in the plant or part of the plant comprises a detectable amount of said at least one PUFA, wherein the PUFA PKS system is a bacterial PUFA PKS system that produces PUFAs at temperatures of at least about 25° C., and wherein the bacterial PUFA PKS system comprises:
a) at least one enoyl ACP-reductase (ER) domain;
b) at least six acyl carrier protein (ACP) domains;
c) at least two β-keto acyl-ACP synthase (KS) domains;
d) at least one acyltransferase (AT) domain;
e) at least one ketoreductase (KR) domain;
f) at least two FabA-like β-hydroxy acyl-ACP dehydrase (DH) domains;
g) at least one chain length factor (CLF) domain;
h) at least one malonyl-CoA:ACP acyltransferase (MAT) domain; and
i) at least one 4′-phosphopantetheinyl transferase (PPTase) domain.
26 . The plant or part of a plant of claim 25 , wherein the PUFA PKS system is from a microorganism selected from the group consisting of: Shewanella olleyana Australian Collection of Antarctic Microorganisms (ACAM) strain number 644 ; Shewanella japonica ATCC strain number BAA-316, and a mutant of any of said microorganisms.
27 . The plant or part of a plant of claim 25 , wherein the nucleic acid sequences encoding the PUFA PKS system hybridize under stringent hybridization conditions to the genes encoding the PUFA PKS system from a microorganism selected from the group consisting of: Shewanella olleyana Australian Collection of Antarctic Microorganisms (ACAM) strain number 644; or Shewanella japonica ATCC strain number BAA-316, or a mutant of any of said microorganisms.
28 . The plant or part of a plant of claim 12 , wherein the PUFA PKS system further comprises a phosphopantetheinyl transferase (PPTase).
29 . An oilseed plant, or part of the oilseed plant, that produces mature seeds in which the total seed fatty acid profile comprises at least 1.0% by weight of at least one polyunsaturated fatty acid having at least twenty carbon atoms and at least four carbon-carbon double bonds, and wherein the total fatty acid profile in the plant or part of the plant contains less than 5% in total of all of the following PUFAs: gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds.
30 . An oilseed plant, or part of the oilseed plant, that produces mature seeds in which the total seed fatty acid profile comprises at least 1.0% by weight of at least one polyunsaturated fatty acid having at least twenty carbon atoms and at least four carbon-carbon double bonds, and wherein the total fatty acid profile in the plant or part of the plant contains less than 1% of gamma-linolenic acid (GLA; 18:3, n-6).
31 . The plant or part of a plant of claim 1 , wherein the at least one PUFA has at least twenty carbons and five or more carbon-carbon double bonds.
32 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is selected from the group consisting of: DHA (docosahexaenoic acid (C22:6, n-3)), ARA (eicosatetraenoic acid or arachidonic acid (C20:4, n-6)), DPA (docosapentaenoic acid (C22:5, n-6 or n-3)), and EPA (eicosapentaenoic acid (C20:5, n-3).
33 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is selected from the group consisting of: DHA (docosahexaenoic acid (C22:6, n-3)), DPA (docosapentaenoic acid (C22:5, n-6 or n-3)), and EPA (eicosapentaenoic acid (C20:5, n-3).
34 . The plant or part of a plant of claim 12 , wherein the at least one PUFA is selected from the group consisting of: DHA (docosahexaenoic acid (C22:6, n-3)), ARA (eicosatetraenoic acid or arachidonic acid (C20:4, n-6)), DPA (docosapentaenoic acid (C22:5, n-6 or n-3)), EPA (eicosapentaenoic acid (C20:5, n-3), gamma-linolenic acid (GLA; 18:3, n-6); stearidonic acid (STA or SDA; 18:4, n-3); and dihomo-gamma-linolenic acid (DGLA or HGLA; 20:3, n-6).
35 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is DHA.
36 . The plant or part of a plant of claim 35 , wherein the ratio of EPA:DHA produced by the plant is less than 1:1.
37 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is EPA.
38 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is DHA and DPAn-6.
39 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is EPA and DHA.
40 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is ARA and DHA.
41 . The plant or part of a plant of claim 1 , wherein the at least one PUFA is ARA and EPA.
42 . The plant or part of a plant of claim 1 , wherein the plant is an oilseed plant and wherein the part of the plant is a mature oilseed.
43 . The plant or part of a plant of claim 1 , wherein the plant is a crop plant.
44 . The plant or part of a plant of claim 1 , wherein the plant is a dicotyledonous plant.
45 . The plant or part of a plant of claim 1 , wherein the plant is a monocotyledonous plant.
46 . The plant or part of a plant of claim 1 , wherein the plant is selected from the group consisting of: canola, soybean, rapeseed, linseed, corn, safflower, sunflower and tobacco.
47 . A plant or a part of the plant, wherein the total fatty acid profile in the plant or part of the plant comprises detectable amounts of DHA (docosahexaenoic acid (C22:6, n-3)), and DPA (docosapentaenoic acid (C22:5, n-6), wherein the ratio of DPAn-6 to DHA is 1:1 or greater than 1:1.
48 . The plant or a part of the plant of claim 47 , wherein the total fatty acid profile in the plant or part of the plant contains less than 5% by weight in total of all of the following PUFAs: gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds.
49 . A plant or part of a plant, wherein the plant has been genetically modified with a PUFA PKS system that produces at least one polyunsaturated fatty acid (PUFA), and wherein the total fatty acid profile in the plant or part of the plant comprises a detectable amount of said at least one PUFA, wherein the PUFA PKS system comprises:
a) two enoyl ACP-reductase (ER) domains;
b) eight or nine acyl carrier protein (ACP) domains;
c) two β-keto acyl-ACP synthase (KS) domains;
d) one acyltransferase (AT) domain;
e) one ketoreductase (KR) domain;
f) two FabA-like β-hydroxy acyl-ACP dehydrase (DH) domains;
g) one chain length factor (CLF) domain;
h) one malonyl-CoA:ACP acyltransferase (MAT) domain; and
i) one phosphopantetheinyl transferase (PPTase).
50 . Seeds obtained from the plant or part of plant of claim 1 .
51 . A food product comprising the seeds of claim 50 .
52 . An oil obtained from seeds of the plant of claim 1 .
53 . An oil comprising the fatty acid profile shown in FIG. 2 or FIG. 3 .
54 . An oil blend comprising the oil of claim 52 and another oil.
55 . The oil blend of claim 54 , wherein the another oil is a microbial oil.
56 . The oil blend of claim 54 , wherein the another oil is a fish oil.
57 . An oil comprising the following fatty acids: DHA (C22:6n-3), DPAn-6 (C22:5n-6), oleic acid (C18:1), linolenic acid (C18:3), linoleic acid (C18:2), C16:0, C18.0, C20:0, C20:1n-9, C20:2n-6, C22:1n-9; wherein the oil comprises less than 0.5% of any of the following fatty acids: gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds.
58 . A plant oil comprising at least about 0.5% by weight of at least one polyunsaturated fatty acid (PUFA) having at least twenty carbons and four or more carbon-carbon double bonds, and wherein the total fatty acid profile oil contains less than 5% in total of all of the following PUFAs: gamma-linolenic acid (GLA; 18:3, n-6), PUFAs having 18 carbons and four carbon-carbon double bonds, PUFAs having 20 carbons and three carbon-carbon double bonds, and PUFAs having 22 carbons and two or three carbon-carbon double bonds.
59 . A plant oil comprising detectable amounts of DHA (docosahexaenoic acid (C22:6, n-3)), and DPA (docosapentaenoic acid (C22:5, n-6), wherein the ratio of DPAn-6 to DHA is 1:1 or greater than 1:1.
60 . A food product that contains an oil of claim 52 .
61 . The food product of claim 60 , further comprising the seeds of claim 50 .
62 . A pharmaceutical product that contains an oil of claim 52 .
63 . A method to produce an oil comprising at least one PUFA, comprising recovering an oil from the seeds of claim 50 .
64 . A method to produce an oil comprising at least one PUFA, comprising recovering an oil from the plant or part of plant of claim 1 .
65 . A method to provide a supplement or therapeutic product comprising at least one PUFA to an individual, comprising providing to the individual a plant or part of plant of claim 1.