Coniothyrium minitans xylanase gene Cxy1

• US 6,121,034 A
• Filed: 05/13/1999
• Issued: 09/19/2000
• Est. Priority Date: 05/13/1999
• Status: Expired due to Fees

• Current Assignee: Her Majesty The Queen In Right Of Canada (Government of Canada)
• Original Assignee: Her Majesty The Queen in Right of Canada as represented by the Minister of Agriculture and Agri-Food
• Inventors: Huang, Timothy Y., Huang, Hung Chang, Laroche, Andre J., Cheng, Kuo Joan, Lu, Zhen-Xiang, Frick, Michele M.
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): HUTSON, RICHARD G
• Application Number:
  US09/311,170
• Time in Patent Office: 495 Days
• Field of Search: 435/209, 435/69.1, 435/320.1, 435/3, 435/254.11, 435/255.5, 435/252.3, 536/23.2, 536/23.1
• US Class Current: 435/209
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 9/248 Xylanases
• CPC Class Codes: C12Y 302/01008 Endo-1,4-beta-xylanase (3.2.1.8)

Abstract

The nucleotide sequence of a novel xylanase gene obtained from the soilborne fungus Coniothyrium minitans, and denoted cxy1, is provided. The amino acid sequence of the encoded xylanase enzyme, denoted Cxy1, is also provided.

Claims

1. An isolated nucleic acid molecule encoding a polypeptide having xylanolytic activity, said encoded polypeptide comprising an amino acid sequence selected from the group consisting of:

a) an amino acid sequence depicted in SEQ ID NO;

2 from amino acid 84 to amino acid 384 or a functionally equivalent amino acid sequence having at least 85% homology thereto;

b) an amino acid sequence depicted in SEQ ID NO;

2 from amino acid 22 to amino acid 384 or a functionally equivalent amino acid sequence having at least 85% homology thereto;

or,

c) an amino acid sequence depicted in SEQ ID NO;

2 from amino acid 1 to amino acid 384 or a functionally equivalent amino acid sequence having at least 85% homology thereto.

2. A nucleic acid molecule according to claim 1, wherein said encoded polypeptide comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 84 to amino acid 384 or a functionally equivalent amino acid sequence having at least 85% homology thereto.

3. A nucleic acid molecule according to claim 2, wherein said encoded polypeptide comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 84 to amino acid 384.

4. A nucleic acid molecule according to claim 3, said nucleic acid molecule comprising the nucleotide sequence depicted in SEQ ID NO:

1 from nucleotide 274 to nucleotide 1176.

5. A nucleic acid molecule according to claim 1, wherein said encoded polypeptide comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 22 to amino acid 384 or a functionally equivalent amino acid sequence having at least 85% homology thereto.

6. A nucleic acid molecule according to claim 5, wherein said encoded polypeptide comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 22 to amino acid 384.

7. A nucleic acid molecule according to claim 6, said nucleic acid molecule comprising the nucleotide sequence depicted in SEQ ID NO:

1 from nucleotide 88 to nucleotide 1176.

8. A nucleic acid molecule according to claim 1, wherein said encoded polypeptide comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 1 to amino acid 384 or a functionally equivalent amino acid sequence having at least 85% homology thereto.

9. A nucleic acid molecule according to claim 8, wherein said encoded polypeptide comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 1 to amino acid 384.

10. A nucleic acid molecule according to claim 9, said nucleic acid molecule comprising the nucleotide sequence depicted in SEQ ID NO:

1 from nucleotide 25 to nucleotide 1176.

15. A vector comprising a nucleic acid molecule according to claim 1.

16. A cell other than Coniothyrium minitans, comprising a nucleic acid molecule according to claim 1.

17. A cell according to claim 16, wherein said cell is Pichia pastoris.

18. A method for producing a polypeptide having xylanolytic activity, comprising the steps of:

a) culturing a cell comprising a nucleic acid molecule according to claim 1 under conditions conducive to an expression of a polypeptide encoded by said nucleic acid molecule; and

,

b) recovering said encoded polypeptide from the culture.

19. The method according to claim 18 wherein said cell is other than Coniothyrium minitans.

20. The method according to claim 18 wherein said cell is Pichia pastoris.

11. An isolated nucleic acid molecule encoding a fragment of a polypeptide having xylanolytic activity, said polypeptide comprising a carbohydrate-binding domain, a catalytic domain, and a linker region operably linking said carbohydrate-binding domain and said catalytic domain, said encoded fragment comprising said carbohydrate-binding domain and said linker region and comprising an amino acid sequence depicted in SEQ ID NO:

2 from amino acid 22 to amino acid 83, or a functionally equivalent amino acid sequence having at least 85% homology thereto.

12. A nucleic acid molecule according to claim 11, wherein said encoded fragment of a polypeptide having xylanolytic activity comprises the amino acid sequence depicted in SEQ ID NO:

2 from amino acid 22 to amino acid 83.

13. A nucleic acid molecule according to claim 12, said nucleic acid molecule comprising the nucleotide sequence depicted in SEQ ID NO:

1 from nucleotide 88 to nucleotide 273.

14. A nucleic acid construct comprising a nucleic acid molecule of claim 11 operably linked to a nucleic acid molecule that encodes a catalytic domain of a hemicellulose-degrading enzyme and that is heterologous to said nucleic acid molecule of claim 11.