Diacylgylcerol acyl transferase (DGA1) polynucleotides, and methods of increasing yeast cell lipid production by overexpression of heterologous DGA1
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Abstract
DGA1 catalyzes the final enzymatic step for converting acyl-CoA and 1,2-diacylglycerol to triacylglycerols (TAG) and CoA in yeast. Disclosed are methods for expression in an oleaginous yeast host of polynucleotide sequences encoding DGA1 from Rhodosporidium toruloides, Lipomyces starkeyi, Aurantiochytrium limacinum, Aspergillus terreus, or Claviceps purpurea. Also described herein are engineered recombinant host cells of Yarrowia lipolytica comprising heterologous DGA1 polynucleotides encoding DGA1 proteins, or functionally active portions thereof, having the capability of producing increased lipid production and possessing the characteristic of enhanced glucose consumption efficiency.
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Citations
25 Claims
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1-6. -6. (canceled)
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7. An isolated host cell comprising a heterologous polynucleotide from the group consisting of:
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a) a nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO;
4, SEQ ID NO;
6, SEQ ID NO;
8, SEQ ID NO;
10, SEQ ID NO;
12, SEQ ID NO;
14, SEQ ID NO;
16, or SEQ ID NO;
18, or a complement thereof; andb) a nucleic acid molecule comprising a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence set forth in SEQ ID NO;
4, SEQ ID NO;
6, SEQ ID NO;
8, SEQ ID NO;
10, SEQ ID NO;
12, SEQ ID NO;
14, SEQ ID NO;
16, or SEQ ID NO;
18, or a complement thereof. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A method of increasing lipid content in a transformed host cell comprising:
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a) providing a transformed host cell comprising; i. a heterologous polynucleotide selected from the group consisting of; 1. a nucleotide sequence set forth in SEQ ID NO;
4, SEQ ID NO;
6, SEQ ID NO;
8, SEQ ID NO;
10, SEQ ID NO;
12, SEQ ID NO;
14, SEQ ID NO;
16, or SEQ ID NO;
18, or a complement thereof; and2. a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence set forth in SEQ ID NO;
4, SEQ ID NO;
6, SEQ ID NO;
8, SEQ ID NO;
10, SEQ ID NO;
12, SEQ ID NO;
14, SEQ ID NO;
16, or SEQ ID NO;
18, or a complement thereof;said polynucleotide encoding a DGA1 polypeptide selected from the group consisting of;
i) amino acid sequence set forth in SEQ ID NO;
3, SEQ ID NO;
5, SEQ ID NO;
7, SEQ ID NO;
9, SEQ ID NO;
1 1, SEQ ID NO;
13, SEQ ID NO;
15, or SEQ ID NO;
17, or a biologically-active portion thereof; and
ii) a polypeptide having at least 80% sequence identity to the amino acid sequence set forth in SEQ ID NO;
3, SEQ ID NO;
5, SEQ ID NO;
7, SEQ ID NO;
9, SEQ ID NO;
1 1, SEQ ID NO;
13, SEQ ID NO;
15, or SEQ ID NO;
17, or a biologically-active portion thereof;b) growing the cell of step (a) under conditions whereby the nucleic acid molecule encoding DGA1 polypeptide is expressed, resulting in the production of lipids; and c) recovering the lipids of step (b). - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25)
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Specification