Fungal cellulase composition containing alkaline CMC-endoglucanase and essentially no cellobiohydrolase and method of making thereof

• US 5,776,757 A
• Filed: 05/30/1995
• Issued: 07/07/1998
• Est. Priority Date: 03/24/1988
• Status: Expired due to Term

• Current Assignee: Novozymes A/S (Novozymes)
• Original Assignee: Novo Nordisk A/S
• Inventors: Levring, Kirsten B.o slashed.egh, Schulein, Martin
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Slobodyansky, Elizabeth
• Application Number:
  US08/452,943
• Time in Patent Office: 1,134 Days
• Field of Search: 435/209, 435/320.1, 435/252.3, 435/254.11, 435/254.3, 435/252.33, 435/252.35, 435/252.31, 435/254.7, 435/911, 435/929, 435/69.1, 536/23.2, 536/23.74
• US Class Current: 435/209
• CPC Class Codes: C11D 3/38645 containing cellulase
• 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: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: D06M 16/003 with enzymes or microorganisms
• CPC Class Codes: D06M 2101/06 cellulosic
• CPC Class Codes: D06M 2200/50 Modified hand or grip properties; Softening compositions
• CPC Class Codes: Y10S 435/911 using fungi
• CPC Class Codes: Y10S 435/929 Fusarium

Abstract

A cellulase preparation useful for reducing the harshness of cotton-containing fabrics and for reducing the rate at which such fabrics become harsh comprises about 40% or more (based on the total protein content) of an endoglucanase component with a pH optimum of about 7.5-10.0 with a high CMC-endoase activity and affinity towards cellulose and essentially no cellobiohydrolase activity. The cellulase composition is endogenous to a strain of Humicola, Myceliophthora, or Fusarium. A method for the recombinant production of said cellulase preparation is provided.

Claims

1. A method for producing a cellulase preparation which comprises a substantially homogeneous endoglucanase, comprising:

(a) introducing a recombinant DNA vector carrying a first DNA sequence encoding the endoglucanase operably linked to control sequences permitting the expression of said first DNA sequence into a host cell;

(b) cultivating the host cell in a culture medium under conditions permitting the expression of the endoglucanase; and

(c) recovering the endoglucanase from the culture medium;

wherein the endoglucanase is endogenous to a strain of Myceliophthora, Fusarium or Humicola and

(i) has a pH optimum of about 7.5-10.0;

(ii) exhibits a carboxymethylcellulose (CMC) endoase activity of at least about 10 CMC-endoase units per mg of total protein;

(iii) exhibits a cellulose affinity viscosity unit (CAVU) activity of at least about 50% at a pH of 7.0-12.0; and

(iv) has essentially no cellobiohydrolase activity.

2. The method according to claim 1, wherein the host cell is an Aspergillus cell.

3. The method according to claim 2, wherein the host cell is an Aspergillus oryzae or Aspergillus niger cell.

4. The method according to claim 1, wherein the host cell is a bacterial cell.

5. The method according to claim 4, wherein the host cell is a Bacillus, Streptomyces or E. coli cell.

6. The method according to claim 1, wherein the endoglucanase exhibits a CMC-endoase activity of at least about 15 CMC-endoase units per mg of total protein.

7. The method according to claim 6, wherein the endoglucanase exhibits a CMC-endoase activity of at least about 50 CMC-endoase units per mg of total protein.

8. The method according to claim 1, wherein the endoglucanase exhibits a CAVU activity in excess of about 65%.

9. The method according to claim 8, wherein the endoglucanase exhibits a CAVU activity in excess of about 75%.

10. The method according to claim 1, wherein the fungus is Humicola.

11. The method according to claim 10, wherein the fungus is Humicola insolens.

12. The method according to claim 1, wherein the fungus is Fusarium.

13. The method according to claim 12, wherein the fungus is Fusarium oxysporum.

14. The method according to claim 1, wherein the fungus is Myceliophthora.

15. The method according to claim 1, wherein the endoglucanase has a molecular weight of about 65 kilodaltons as determined by denaturing gel electrophoresis.

16. The method according to claim 15, wherein the fungus is Humicola.

17. The method according to claim 16, wherein the fungus is Humicola insolens.

18. The method according to claim 1, wherein the endoglucanase has

(e) an isoelectric point of about 8.5-9.5;

and

(f) an immunological reaction with a polyspecific antibody raised against cellulase derived from Humicola insolens DSM 1800.

19. The method according to claim 1, wherein the endoglucanase has a molecular weight of about 80 kilodaltons as determined by denaturing gel electrophoresis.

20. The method according to claim 19, wherein the fungus is Fusarium.

21. The method according to claim 20, wherein the fungus is Fusarium oxysporum.