Genes that improve tolerance to lignocellulosic toxins when overexpressed in yeast
The present invention provides isolated gene sequences useful in increasing lignocellulosic toxin tolerance in yeast. Such engineered yeast are useful in methods of biofuel production, particularly ethanol production. Methods of bioengineering recombinant yeast with increased lignocellulosic toxin tolerance are also provided.
1. A recombinant vector comprising: (a) the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3; and (b) a promoter operably-linked to the nucleotide sequence of (a); wherein overexpression in yeast of said nucleotide sequence provides increased tolerance to lignocellulosic toxins relative to a control yeast lacking overexpression of the nucleotide sequence.
2. The recombinant vector of claim 1 , wherein said vector includes heterologous nucleotide sequences that stably maintain the vector at a high copy number when transformed into yeast.
3. The recombinant vector of claim 1 , wherein the promoter is a heterologous promoter.
4. A recombinant yeast comprising the recombinant vector of claim 1 .
5. The recombinant yeast of claim 4 , wherein the recombinant yeast is of the genus Saccharomyces.
6. The recombinant yeast of claim 5 , wherein the recombinant yeast is of the species Saccharomyces cerevisiae.
7. The recombinant yeast of claim 4 , wherein the recombinant vector is an extrachromosomal vector stably maintained in the recombinant yeast.
8. The recombinant yeast of claim 4 , wherein the recombinant vector is stably maintained at a high copy number in the recombinant yeast.
9. The recombinant yeast of claim 4 , wherein the recombinant vector is integrated into a chromosome of the recombinant yeast.
10. A recombinant Saccharomyces cerevisiae strain, comprising: an isolated nucleotide sequence encoding and overexpressing an adenylylsulfate kinase (MET14), protein folding protein folding co-chaperone (MDJ1), or C3 sterol dehydrogenase (ERG26); wherein the isolated nucleotide sequence is contained in an extrachromosomal vector maintained at a high copy number in the strain, and said strain exhibits increased tolerance to lignocellulosic toxins relative to a control strain lacking the isolated nucleotide sequence.