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US 11,987,819 B2Granted
Method of increasing omega-3 polyunsaturated fatty acids production in microalgae
AssigneeDsm IP Assets B.V.
InventorsAnne-Cecile V. Bayne, Ross E. Zirkle
Life of this patent
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Abstract
The subject disclosure features, in one aspect, a method for producing lipids enriched for EPA, comprising modifying a microalga to increase expression of PFA1, and/or PFA3, and culturing the modified microalga under conditions which allow the expression of PFA1, and/or PFA3, wherein lipids enriched for EPA are produced. Also featured is a recombinant microalga in which PFA1, and/or PFA3, is overexpressed. Such recombinant microalgae have been demonstrated herein to produce very favorable fatty acid lipid profiles (e.g., increased levels of EPA, increased ratio of EPA:DHA, decreased levels of DPA n-6, etc.).
Claims13
A recombinant microalga that has been modified to express increased levels of polyunsaturated fatty acid synthase, comprising:
- (a) a first gene encoding a polyunsaturated fatty acid synthase subunit 1 (PFA1) which comprises a first amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 2, wherein the first amino acid sequence comprises a β-ketoacyl-ACP synthase (KS) domain having at least 95% sequence identity to SEQ ID NO:
- 8, a malonyl-CoA:ACP acyltransferase (MAT) domain having at least 95% sequence identity to SEQ ID NO:
- 10, an acyl carrier protein (ACP) domain having at least 95% sequence identity to SEQ ID NO:
- 12, 14, 16, 18, 20, 22, or 24, a β-ketoacyl-ACP reductase (KR) domain of SEQ ID NO:
- 26 and a FabA-like β-hydroxyacyl-ACP dehydrase (DH) domain having at least 95% sequence identity to SEQ ID NO:
- 28;
- (b) a second gene encoding a polyunsaturated fatty acid synthase subunit 2 (PFA2) which comprises a second amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 4, wherein the second amino acid sequence comprises a KS domain having at least 95% sequence identity to SEQ ID NO:
- 30, a chain length factor (CLF) domain having at least 95% sequence identity to SEQ ID NO:
- 32, an acyltransferase (AT) domain having at least 95% sequence identity to SEQ ID NO:
- 34 and an enoyl-ACP reductase (ER) domain having at least 95% sequence identity to SEQ ID NO:
- 36;
- and
- (c) a third gene encoding a polyunsaturated fatty acid synthase subunit 3 (PFA3) which comprises a third amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 6, wherein the third amino acid sequence comprises a DH domain having at least 95% sequence identity to SEQ ID NO:
- 38 or 40 and an ER domain having at least 95% sequence identity to SEQ ID NO:
- 42,
- wherein the recombinant microalga is a recombinant Schizochytrium,
- wherein the recombinant microalga comprises multiple copies of the first and/or third gene but only a single copy of the second gene, and
- wherein the recombinant microalga produces lipids with an increased level of eicosapentaenoic acid (EPA) compared to an unmodified microalga.
18 more elements
A recombinant plant that has been modified to express increased levels of polyunsaturated fatty acid synthase, comprising:
- (a) a first gene encoding a polyunsaturated fatty acid synthase subunit 1 (PFA1) which comprises a first amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 2, wherein the first amino acid sequence comprises a β-ketoacyl-ACP synthase (KS) domain having at least 95% sequence identity to SEQ ID NO:
- 8, a malonyl-CoA:ACP acyltransferase (MAT) domain having at least 95% sequence identity to SEQ ID NO:
- 10, an acyl carrier protein (ACP) domain having at least 95% sequence identity to SEQ ID NO:
- 12, 14, 16, 18, 20, 22, or 24, a β-ketoacyl-ACP reductase (KR) domain having at least 95% sequence identity to SEQ ID NO:
- 26 and a FabA-like p-hydroxyacyl-ACP dehydrase (DH) domain having at least 95% sequence identity to SEQ ID NO:
- 28;
- (b) a second gene encoding a polyunsaturated fatty acid synthase subunit 2 (PFA2) which comprises a second amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 4, wherein the second amino acid sequence comprises a KS domain having at least 95% sequence identity to SEQ ID NO:
- 30, a chain length factor (CLF) domain having at least 95% sequence identity to SEQ ID NO:
- 32, an acyltransferase (AT) domain having at least 95% sequence identity to SEQ ID NO:
- 34 and a enoyl-ACP reductase (ER) domain having at least 95% sequence identity to SEQ ID NO:
- 36;
- and
- (c) a third gene encoding a polyunsaturated fatty acid synthase subunit 3 (PFA3) which comprises a third amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 6, wherein the third amino acid sequence comprises a DH domain having at least 95% sequence identity to SEQ ID NO:
- 38 or 40 and an ER domain having at least 95% sequence identity to SEQ ID NO:
- 42,
- wherein the recombinant plant further comprises a polynucleotide encoding at least one phosphopantetheinyl transferase (PPTase),
- wherein the recombinant plant comprises multiple copies of the first and/or third transgene but only a single copy of the second transgene, and
- wherein the recombinant plant produces lipids with an increased level of eicosapentaenoic acid (EPA) compared to an unmodified plant.
18 more elements
Dependencies are inferred from the claim text.