Genetically modified strain of S. cerevisiae engineered to ferment xylose and arabinose

Patent metadata

Field	Value
Publication number	US-9598689-B2
Application number	US-201214356510-A
Country	US
Kind code	B2
Filing date	Nov 9, 2012
Priority date	Nov 10, 2011
Publication date	Mar 21, 2017
Grant date	Mar 21, 2017

Abstract

The present invention provides a microorganism capable of fermenting arabinose to a desired product such as ethanol. In some embodiments, the organism is also capable of fermenting xylose. In some embodiments, the organism is capable of fermenting arabinose and xylose, and expresses one or more cellulases.

First claim

What is claimed is:

1. A recombinant eukaryotic host cell comprising a heterologous polynucleotide encoding an arabinose isomerase (AI), a heterologous polynucleotide encoding a ribulokinase (RK) and a heterologous polynucleotide encoding a ribulose 5-phosphate epimerase (R5PE), wherein one or more of the AI, RK and R5PE comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO:8.

2. The recombinant eukaryoiic host cell of either claim 1 , wherein
a) the AI comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 6;
b) the RK comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 7; and
c) the R5PE comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 8.

3. The recombinant eukaryotic host cell of either claim 1 , wherein expression of the heterologous polynucleotide confers an ability to ferment arabinose to the recombinant host cell.

4. The recombinant eukaryotic host cell of either claim 1 , further comprising a heterologous polynucleotide encoding a xylose isomerase (XI).

5. The recombinant eukaryotic host ceil of claim 4 , wherein the XI is derived from an XI of B. thetaiotamicron.

6. The recombinant eukaryotic host cell of either of claim 4 , wherein the XI comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 23 or SEQ ID NO: 25.

7. The recombinant eukaryotic host cell of either claim 1 , wherein the host cell is a yeast cell.

8. The recombinant eukaryotic host cell of claim 7 , wherein the yeast cell is selected from the group consisting of Saccharomyces cerevisiae, Schizzosaccharomyces pombe, Candida albicans, Pichia pastoris, Pichia stipitis, Yarrowia lipolytica, Hansenula polymorpha, Phaffia rhodozyma, Candida utilis, Arxula adeninivorans, Debaryomyces hansenii, Debaryomyces polymorphous, Schizosaccharomyces pombe and Schwanniomyces occidentalis.

9. The recombinant eukaryotic host cell of claim 7 , wherein the yeast cell comprises a heterologous sequence encoding a xylulokinase, ribulose 5-phosphate isomerase, ribulose 5-phophate epimerase, transketolase and transaldolase, and wherein the yeast cell does not express an aldose reductase that is capable of catalyzing the conversion of xylose to xylitol.

10. The recombinant eukaryotic host cell of any one of either claim 1 , wherein the host cell is capable of fermenting xylose, arabinose, or a combination thereof, and producing a fermentation product.

11. The recombinant eukaryotic host cell of claim 10 , wherein the host cell is capable of fermenting arabinose from a cellulosic substrate.

12. The recombinant eukaryotic host cell of claim 10 , wherein the fermentation product is selected from the group consisting of ethanol, lactic acid, hydrogen, butyric acid, acetone, and butanol.

13. The recombinant eukaryotic host cell of either claim 1 , wherein the host cell is an industrial strain exhibiting high ethanol tolerance.

14. The recombinant eukaryotic host cell of claim 13 , wherein the host cell further exhibits high temperature tolerance.

15. The recombinant eukaryotic host cell of either claim 1 , wherein the host cell produces an ethanol yield of at least about 10 g/l ethanol after 24 hours of fermentation from a medium containing 20 g/l xylose and 21 g/l arabinose.

16. The recombinant eukaryotic host cell of either claim 1 , wherein the host cell produces an ethanol yield of at least about 13 g/l ethanol after 24 hours of fermentation from a medium containing 20 g/l glucose and 21 g/l arabinose.

17. The recombinant eukaryotic host cell of either claim 1 , wherein the host cell produces an ethanol yield of at least about 15 g/l ethanol after 24 hours of fermentation from a medium containing 10 g/l glucose, 10 g/l xylose and 21 g/l arabinose.

18. The recombinant eukaryotic host cell of either claim 1 , wherein the host cell further comprises one or more heterologous polynucleotides encoding a cellulase.

19. The recombinant eukaryotic host cell of claim 18 , wherein the one or more cellulases is selected from the group consisting of endoglucanases, exoglucanases, and .beta.-glucosidases.

20. The recombinant eukaryotic host cell of claim 19 , wherein the host cell comprises: (a) a first heterologous polynucleotide that encodes an endoglucanase; (b) a second heterologous polynucleotide that encodes a .beta.-glucosidase; (c) a third heterologous polynucleotide that encodes a first cellobiohydrolase; and, (d) a fourth heterologous polynucleotide that encodes a second cellobiohydrolase.

21. The recombinant eukaryotic host cell of claim 20 , wherein (a) the first heterologous polynucleotide that encodes an endoglucanase is derived from A. fumigatus ; (b) the second heterologous polynucleotide that encodes a .beta.-glucosidase is derived from S. fibuligera ; (c) the third heterologous polynucleotide that encodes a first cellobiohydrolase is derived from T. emersonii ; and, (d) the fourth heterologous polynucleotide that encodes a second cellobiohydrolase is derived from C. lucknowense.

22. The recombinant host cell of any one of claims 1 , wherein at least one of the heterologous polynucleotides is integrated into the genome of the host cell.

23. A composition comprising a carbon source and the recombinant eukaryotic host cell of any one of claims 1 , wherein the carbon source is a cellulosic substrate that contains at least about 1% arabinose.

Assignees

• Lallemand Hungary Liquidity Man Llc

Inventors

• Argyros D Aaron
• Caiazza Nicky
• Barrett Trisha F
• Warner Anne K

Classifications

• C12N9/90 Primary

  Isomerases (5.) · CPC title

• C07K14/38

  from Aspergillus · CPC title

• C07K14/39

  from yeasts · CPC title

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• C12N9/1205

  Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases · CPC title