SYNTHESIS OF LONG-CHAIN POLYUNSATURATED FATTY ACIDS BY RECOMBINANT CELL
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Abstract
The present invention relates to methods of synthesizing long-chain polyunsaturated fatty acids, especially eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid, in recombinant cells such as yeast or plant cells. Also provided are recombinant cells or plants which produce long-chain polyunsaturated fatty acids. Furthermore, the present invention relates to a group of new enzymes which possess desatorase or elongase activity that can be used in methods of synthesizing long-chain polyunsaturated fatty acids.
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Citations
235 Claims
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1-221. -221. (canceled)
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222. A process for producing fish comprising:
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a) obtaining feedstuff which comprises oil extracted from transgenic Brassica seeds which comprise eicosapentaenoic acid (EPA) and heterologous polynucleotides which encode enzymes that comprise; i) a Δ
6 desaturase, a Δ
6 elongase and a Δ
5 desaturase;
orii) a Δ
5/Δ
6 bifunctional desaturase and a Δ
5/Δ
6 bifunctional elongase;wherein the heterologous polynucleotides are operably linked to one or more promoters that are capable of directing expression of said polynucleotides in Brassica seed cells, wherein the enzymes encoded by said polynucleotides comprise at least one desaturase which is able to act on an acyl-CoA substrate; and b) administering the feedstuff to fish, thereby producing the fish. - View Dependent Claims (223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233)
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234. A process for producing fish comprising:
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a) obtaining feedstuff which comprises oil extracted from transgenic Brassica seeds which comprise docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) in an esterified form as part of triacylglycerols in the seeds, and which comprise i) a polynucleotide which encodes a Δ
5 elongase operably linked to a promoter which directs expression of the polynucleotide in Brassica seed cells;ii) a polynucleotide which encodes a Δ
4 desaturase whose amino acid sequence is at least 99% identical to the sequence set forth as SEQ ID NO;
33, operably linked to a promoter which directs expression of the polynucleotide in the Brassica seed cells; andiii) an exogenous desaturase which desaturates an acyl-CoA substrate; and b) administering the feedstuff to fish, thereby producing the fish.
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235. A process for producing fish comprising:
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a) obtaining feedstuff which comprises oil extracted from transgenic Brassica seeds which comprise docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) in an esterified form as part of triacylglycerols in the seeds, and heterologous polynucleotides which encode enzymes that comprise; i) a Δ
5 elongase;ii) a Δ
6 elongase which has the amino acid sequence set forth as SEQ ID NO;
31;iii) a Δ
4 desaturase whose amino acid sequence is at least 99% identical to the sequence set forth as SEQ ID NO;
33;iv) a Δ
5 desaturase which has the amino acid sequence set forth as SEQ ID NO;
18; andv) a Δ
6 desaturase;wherein the heterologous polynucleotides are operably linked to one or more promoters that are capable of directing expression of said polynucleotides in Brassica seed cells, wherein the Brassica seeds comprise an exogenous desaturase which desaturates an acyl-CoA substrate; and b) administering the feedstuff to fish, thereby producing the fish.
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Specification