High eicosapentaenoic acid producing strains of Yarrowia lipolytica
First Claim
1. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
6 desaturase;
b) at least one gene encoding C18/20 elongase;
c) at least one gene encoding Δ
5 desaturase; and
, d) at least one gene encoding Δ
17 desaturase.
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Accused Products
Abstract
Engineered strains of the oleaginous yeast Yarrowia lipolytica capable of producing greater than 25% eicosapentaenoic acid (EPA, an ω-3 polyunsaturated fatty acid) in the total oil fraction are described. These strains comprise various chimeric genes expressing heterologous desaturases, elongases and acyltransferases and optionally comprise various native desaturase and acyltransferase knockouts to enable synthesis and high accumulation of EPA. Production host cells are claimed, as are methods for producing EPA within said host cells.
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Citations
53 Claims
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1. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
6 desaturase;
b) at least one gene encoding C18/20 elongase;
c) at least one gene encoding Δ
5 desaturase; and
,d) at least one gene encoding Δ
17 desaturase. - View Dependent Claims (5, 6, 7, 8, 9, 11, 12, 13, 15, 19, 20, 21, 22, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52)
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2. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
15 desaturase;
b) at least one gene encoding Δ
6 desaturase;
c) at least one gene encoding C18/20 elongase; and
,d) at least one gene encoding Δ
5 desaturase. - View Dependent Claims (16, 17, 18)
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3. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
9 elongase;
b) at least one gene encoding Δ
8 desaturase;
c) at least one gene encoding Δ
5 desaturase; and
,d) at least one gene encoding Δ
17 desaturase. - View Dependent Claims (10, 14)
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4. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
15 desaturase;
b) at least one gene encoding Δ
9 elongase;
c) at least one gene encoding Δ
8 desaturase; and
,d) at least one gene encoding Δ
5 desaturase.
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23. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
6 desaturase;
b) at least one gene encoding C18/20 elongase;
c) at least one gene encoding Δ
5 desaturase;
d) at least one gene encoding Δ
17 desaturase;
e) at least one gene encoding C16/18 elongase; and
,f) at least one gene encoding Δ
12 desaturase; and
wherein the background Yarrowia sp. is devoid of any native gene encoding an enzyme selected from the group consisting of;
peroxisomal acyl-CoA oxidase ACO3 (Pox3-), lipase 1 (Lip1-) and Δ
12 desaurase.
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24. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
9 elongase;
b) at least one gene encoding Δ
8 desaturase;
c) at least one gene encoding Δ
5 desaturase;
d) at least one gene encoding Δ
17 desaturase;
e) at least one gene encoding C16/18 elongase; and
,f) at least one gene encoding Δ
12 desaturase; and
wherein the background Yarrowia sp. is devoid of any native gene encoding an enzyme selected from the group consisting of;
acyl-CoA oxidase 2 (Pox2-), and saccharopine dehydrogenase (Lys5-).
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25. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one gene encoding Δ
15 desaturase;
b) at least one gene encoding Δ
9 elongase;
c) at least one gene encoding Δ
8 desaturase;
d) at least one gene encoding Δ
5 desaturase; and
,e) at least one gene encoding Δ
12 desaturase.
- -3/ω
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26. A recombinant production host cell for the production of eicosapentaenoic acid comprising a background Yarrowia sp. comprising a gene pool comprising the following genes of the ω
- -3/ω
-6 fatty acid biosynthetic pathway;
a) at least one set of genes selected from the group consisting of;
(1) at least one gene encoding Δ
6 desaturase;
at least one gene encoding C18/20 elongase;
at least one gene encoding Δ
5 desaturase; and
, at least one gene encoding Δ
17 desaturase;
(2) at least one gene encoding Δ
9 elongase;
at least one gene encoding Δ
8 desaturase;
at least one gene encoding Δ
5 desaturase; and
, at least one gene encoding Δ
17 desaturase;
(3) at least one gene encoding Δ
15 desaturase;
at least one gene encoding Δ
6 desaturase;
at least one gene encoding C18/20 elongase; and
, at least one gene encoding Δ
5 desaturase;
(4) at least one gene encoding Δ
15 desaturase;
at least one gene encoding Δ
9 elongase;
at least one gene encoding Δ
8 desaturase;
at least one gene encoding Δ
5 desaturase; and
,b) at least one gene encoding an enzyme selected from the group consisting of;
(1) Δ
12 desaturase;
(2) Δ
9 desaturase;
(3) C14/16 elongase;
(4) C16/18 elongase; and
,c) at least one gene encoding an enzyme selected from the group consisting of;
(i) diacylglycerol acyltransferase selected from the group consisting of DGAT1, DGAT2 and PDAT;
(ii) acyl-CoA;
1-acyl lysophosphatidylcholine acyltransferase (LPCAT);
(iii) glycerol-3-phosphate acyltransferase (G PAT);
(iv) lysophosphatidic acid acyltransferase (LPAAT);
(v) phospholipase C; and
,(vi) phospholipase A2;
wherein;
(1) the background Yarrowia sp. is devoid of any native gene encoding a polypeptide having Δ
12 desaturase activity; and
,(2) the background Yarrowia sp. is devoid of any native gene encoding an enzyme selected from the group consisting of lipase 1 (Lip1-), peroxisomal acyl CoA oxidase ACO3 (Pox3-), acyl-CoA oxidase 2 (Pox2-), orotidine-5′
-phosphate decarboxylase (Ura3-), saccharopine dehydrogenase (Lys5-), lipase 2 (Lip2-) and isopropyl malate dehydrogenase (Leu2-).
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53. A recombinant production host useful for the production of eicosapentaenoic acid selected from the group consisting of;
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Yarrowia lipolytica Y2047 having the ATCC designation ATCC ______;
Yarrowia lipolytica Y2201 having the ATCC designation ATCC ______; and
Yarrowia lipolytica Y2096 having the ATCC designation ATCC ______.
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Yarrowia lipolytica Y2047 having the ATCC designation ATCC ______;
Specification