Bacillus 029cel cellulase

• US 7,604,974 B2
• Filed: 04/28/2004
• Issued: 10/20/2009
• Est. Priority Date: 04/29/2003
• Status: Expired due to Fees

• Current Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Grant, William D., Jones, Brian E., Heaphy, Shaun, Grant, Susan
• Primary Examiner(s): Nashed; Nashaat T
• Assistant Examiner(s): CHOWDHURY, IQBAL HOSSAIN
• Application Number:
  US10/549,943
• Publication Number:
  US 20070172916A1
• Time in Patent Office: 2,001 Days
• Field of Search: 435/69.1, 435/105, 435/161, 435/209, 435/252.31, 536/23.2, 510/320
• US Class Current: 435/209
• CPC Class Codes: C12N 9/2437 Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
• CPC Class Codes: C12P 19/14 produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase , e.g. by cellulase, hemicellulase
• CPC Class Codes: C12P 7/06 Ethanol, i.e. non-beverage
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The present invention provides a novel cellulase nucleic acid sequence, designated 029cel, and the corresponding 029cel amino acid sequence. The invention also provides expression vectors and host cells comprising a nucleic acid sequence encoding 029cel, recombinant 029cel proteins and methods for producing the same.

MNFSLPIRFF GIGPTAGFFL LLMGSSALVA 50 STHTDKIRID QFGYPADATK VAVIADPQIG WNSAESYSPG ATLEVRRVND 100 GVVVFSGSPV PWNGGAIHIQ SGDRVWWFDF TVVAEPGHYR IHDPANNTHS 150 DSFAIGADVY DVVLREAVRM FFYQRSGFAK EVPYAHANWA DAASHPQDVA 200 SRPIWDMGNA SLERDLSGGW FDAGDFNKYS EWTGRVILEL LLAYQGRPDV 250 FTDDFGIPES GNGVPDLLDE VKWGMDWLLR MQEPSGAILG KVSVTGHQSA 300 SPPSTDTHPR YYGPVSTEAT AMAAAAFALG ATVFESVGMS DYAVTLESAA 350 IAAWNWTMVH PHVPFDNTGF ASVSPSRNAH DTLANRVMAA AMLFERTGGA 400 VYRDFFDVRY LDMEPVQWWY FFPFQGELQK ALAHYTTLPG ATPSVSADIR 450 NRMAASINGG EFLGAWNNQT DAYRAYLKDQ DYTWGSNKTK SQAGFFFEGV 500 RRLGLNPADA AAHRDAAMGY LHYLHGVNPM GMVYLSNMYA SGADRAANEI 550 YHHWFRDGRT GTMPSLHSTV PLLVFFRAGP NAQIQRKHSG DPRPTRAKSL 581 P

Claims

1. An isolated polynucleotide selected from the group consisting of:

(a) a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO;

2; and

(b) a nucleic acid sequence that encodes a polypeptide having at least 90% sequence identity to the amino acid sequence presented in FIG. 3 (SEQ ID NO;

3);

wherein said isolated polynucleotide encodes a polypeptide having cellulase activity.

2. The isolated polynucleotide of claim 1 wherein the polynucleotide is selected from the group mRNA, DNA, cDNA, and genomic DNA.

3. The isolated polynucleotide of claim 2, wherein said polynucleotide is an RNA molecule.

4. The isolated polynucleotide of claim 1 encoding an enzyme having cellulase activity, wherein the enzyme is isolated from a Bacillus source.

5. The nucleic acid sequence of claim 1, wherein the nucleic acid sequence encodes a polypeptide having the amino acid sequence presented in FIG. 3 (SEQ ID NO:

3).

6. An expression construct comprising the polynucleotide of claim 1.

7. An expression vector comprising polynucleotide of claim 1 operably linked to control sequences recognized by a host cell transformed with the vector.

8. An expression vector according to claim 7 further comprising a regulatory polynucleotide sequence including a promoter sequence derived from a glucose isomerase gene of Actinoplanes and a signal sequence derived from a Streptomyces cellulase gene.

9. A vector comprising the expression construct of claim 6.

10. An isolated host cell transformed with the vector of claim 7.

11. The host cell of claim 10, which is a prokaryotic cell.

12. The host cell of claim 10, which is a eukaryotic cell.

16. A method of producing a cellulase comprising the steps of:

(a) culturing the host cell according to claim 10 in a suitable culture medium under suitable conditions to produce the cellulase;

(b) obtaining said produced cellulase.

17. A purified enzyme having cellulase activity prepared by the method of claim 16.

18. The method of claim 16 wherein the host cell is a filamentous fungi or yeast cell.

19. The method of claim 16 wherein the host cell is a bacterium.

20. The method of claim 19 wherein the bacterium is a Streptomyces.

13. An isolated polypeptide with cellulase activity, comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence presented in FIG. 3 (SEQ ID NO:

3).

14. The substantially purified polypeptide according to claim 13 which is obtainable from a Bacillus.

15. The polypeptide of claim 13, wherein the polypeptide has the amino acid sequence presented in FIG. 3 (SEQ ID NO:

3).

21. A detergent composition comprising a polypeptide of claim 13.

22. A detergent composition comprising a surfactant and a cellulase according to claim 13.

23. The detergent according to claim 21, wherein said detergent is a laundry detergent.

24. The detergent according to claim 21, wherein said detergent is a household detergent.

25. A method of treating wood pulp comprising contacting said wood pulp with a cellulase according to claim 13.