Yeast 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.
Claims17
A genetically modified yeast cell comprising the following modifications:
- (a) a first nucleic acid encoding a Mus musculus fatty acid reductase (FAR)(MmFAR1) or Tyto alba FAR1 (TaFAR1) operatively linked to a promoter, and (b) a second nucleic acid encoding acetyl-CoA carboxylase (ACC1) operatively linked to a promoter and (c) (i) a reduced expression of or knocked out for a homolog of fatty acid aldehyde (HFD1), (ii) a reduced expression of or knocked out for an alcohol dehydrogenase (ADH6), (iii) a nucleic acid encoding a mutant of a bottleneck enzyme encoded by ACC1 insensitive to post-transcriptional and post-translational repression (ACC1**) operatively linked to a promoter, (iv) a reduced expression of or knocked out for a glutamate dehydrogenase (GDH1), or (v) a nucleic acid encoding a fatty acid-desaturase (OLE1) operatively linked to a promoter;
- wherein the MmFAR1 or TaFAR1, and ACC1 have an increased expression compared to expression of each corresponding MmFAR1 or TaFAR1, and ACC1 in an unmodified yeast cell, and the genetically modified yeast cell, when grown in 1% yeast extract, 2% peptone, and 2% dextrose (YPD) medium, 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:
- (a) introducing a first nucleic acid encoding Mus musculus fatty acid reductase (FAR) MmFAR1 or Tyto alba FAR1 (TaFAR1) operatively linked to a promoter capable of expressing the MmFAR1 or TaFAR1 gene product in the yeast cell;
- (b) introducing a second nucleic acid encoding 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;
- and (c)(i) reducing expression of or knocking out for a homolog of fatty acid aldehyde (HFD1), (ii) reducing expression of or knocking out for an alcohol dehydrogenase (ADH6), (iii) introducing a nucleic acid encoding a mutant of a bottleneck enzyme encoded by ACC1 insensitive to post-transcriptional and post-translational repression (ACC1**) operatively linked to a promoter, (iv) reducing expression of or knocking out for a glutamate dehydrogenase (GDH1), or (v) introducing a nucleic acid encoding a fatty acid-desaturase (OLE1) operatively linked to a promoter;
- wherein the higher transcription activity is higher than each corresponding MmFAR1 or TaFAR1, and ACC1 in an unmodified yeast cell, and the genetically modified yeast cell, when grown in 1% yeast extract, 2% peptone, and 2% dextrose (YPD) medium, is capable of producing at least 1.0 g of fatty alcohol per liter of medium.
Dependencies are inferred from the claim text.