Yeast host cells and methods for producing fatty alcohols
AssigneeThe Regents of the University of California
InventorsLeopold D'Espaux, Jay D. Keasling
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Abstract
The present invention provides for a genetically modified yeast cell comprising at least six or more of the following modifications: increased expression of Mus musculus fatty acid reductase, acetyl-CoA carboxylase, fatty acid synthase 1, fatty acid synthase 2, a mutant of the bottleneck enzyme encoded by ACC1 insensitive to post-transcriptional and post-translational repression, and/or a desaturase encoded by OLE1, and reduced expression of DGA1, HFD1, ADH6, and/or GDH1. The present invention provides a method for constructing the genetically modified yeast cell, and a method for producing a fatty alcohol from the genetically modified yeast cell.
Claims10
A genetically modified yeast cell comprising the following modifications:
- (a) an increased expression of Mus musculus fatty acid reductase (FAR) (MmFAR1);
- (b) an increased expression of acetyl-CoA carboxylase (ACC1);
- (c) an increased expression of fatty acid synthase 1 (FAS1);(d) an increased expression of fatty acid synthase 2 (FAS2);
- (e) a reduced expression of or deleted for diacylglycerol O-acyltransferase (DGA1);
- (f) a reduced expression of or deleted for homolog of fatty acid aldehyde (HFD1);
- (g) a reduced expression of or deleted for alcohol dehydrogenase (ADH6);
- (h) an increased expression of a Saccharomyces cerevisiae ACC1 comprising the mutations S659A and S1157A;
- (i) a reduced expression of or deleted for glutamate dehydrogenase (GDH1);
- and (j) an increased expression of a delta(9) fatty acid desaturase encoded by OLE1;
- wherein the genetically modified yeast cell is capable of producing at least 1.0 g of fatty alcohol per liter of medium.
A method of constructing a genetically modified yeast cell comprising the following steps:
- (a) introducing a first nucleic acid encoding Mus musculus fatty acid reductase (FAR) (MmFAR1), operatively linked to a promoter capable of expressing the MmFAR1 gene product in the yeast cell;
- (b) introducing a second nucleic acid encoding an acetyl-CoA carboxylase (ACC1), operatively linked to a promoter capable of expressing the ACC1 gene product in the yeast cell, or replacing the native promoter of ACC1 with a promoter with a higher transcription activity;
- (c) introducing a third nucleic acid encoding FAS1, operatively linked to a promoter capable of expressing the FAS1 gene product in the yeast cell, or replacing the native promoter of FAS1 with a promoter with a higher transcription activity;
- (d) introducing a fourth nucleic acid encoding FAS2 operatively linked to a promoter capable of expressing the FAS2 gene product in the yeast cell, or replacing the native promoter of FAS2 with a promoter with a higher transcription activity;
- (e) removing all or a portion of a diacylglycerol O-acyltransferase (DGA1) gene, or introducing a nucleic acid into the yeast cell targeting the DGA1 gene or its gene product, wherein the yeast cell has a reduced amount of activity of the DGA1 gene product;
- (f) removing all or a portion of a homolog of fatty acid aldehyde (HFD1) gene, or introducing a nucleic acid into the yeast cell targeting the HFD1 gene or its gene product, wherein the yeast cell has a reduced amount of activity of the HFD1 gene product;
- (g) removing all or a portion of an alcohol dehydrogenase (ADH6) gene, or introducing a nucleic acid into the yeast cell targeting the ADH6 gene or its gene product, wherein the yeast cell has a reduced amount of activity of the ADH6 gene product;
- (h) introducing a fifth nucleic acid encoding a Saccharomyces cerevisiae ACC1 comprising the mutations S659A and S1157A, operatively linked to a promoter capable of expressing the mutant ACC1 gene product in the yeast cell;
- (i) removing all or a portion of a glutamate dehydrogenase (GDH1) gene, or introducing a nucleic acid into the yeast cell targeting the GDH1 gene or its gene product, wherein the yeast cell has a reduced amount of activity of the GDH1 gene product;
- and (j) introducing a sixth nucleic acid encoding a delta(9) fatty acid saturase (OLE1), operatively linked to a promoter capable of expressing the OLE1 gene product in the yeast cell;
- wherein the genetically modified yeast cell is capable of producing at least 1.0 g of fatty alcohol per liter of medium.
Dependencies are inferred from the claim text.