METHOD FOR CELLULASE PRODUCTION

• US 20090061486A1
• Filed: 08/28/2008
• Published: 03/05/2009
• Est. Priority Date: 08/30/2007
• Status: Active Grant

• Current Assignee: Iogen Energy Corporation (Iogen Corporation)
• Original Assignee: Iogen Energy Corporation (Iogen Corporation)
• Inventors: Munkvold, Glenn D., Edwards, Jason B., Foody, Brian E.
• Granted Patent: US 8,093,019 B2
• Time in Patent Office: Days
• US Class Current: 435/72
• 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 19/02 Monosaccharides 2-ketogulonic acid C12P7/60
• CPC Class Codes: C12P 19/04 Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
• 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 21/00 Preparation of peptides or proteins single cell protein C12N1/00
• CPC Class Codes: C12P 2203/00 Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase

Abstract

A fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures with a high proportion of cellulases relative to hemicellulases is provided. The cellulases produced by the process of the invention are further characterized by high specific productivity. The resulting cellulase mixtures comprise at least two times more cellulase than hemicellulase and are useful for the hydrolysis of cellulosic substrated, particularly, pretreated lignocellulosic substrate.

Claims

1. A fermentation process for the production of a cellulase mixture comprising providing a cellulase-secreting host cell of the genus Trichoderma or Hypocrea with a carbon source for cellulase enzyme production, the carbon source comprising a hemicellulose-derived carbohydrate from about 40 weight percent to about 97 weight percent of the total carbon present in the carbon source, and a cellulase-inducing carbohydrate from about 3 weight percent to about 20 weight percent of the total carbon present in the carbon source, for from about 3 to about 30 days, at a temperature of about 20°

C. to about 35°

C., at a pH of from about 3.0 to about 6.5, and obtaining the cellulose mixture.

2. The fermentation process of claim 1, wherein from about 80 weight percent to about 97 weight percent of the total carbon present in the carbon source is the hemicellulose-derived carbohydrate, and wherein from about 8 weight percent to about 15 weight percent of the total carbon present in the carbon source is the cellulase-inducing carbohydrate.

3. The fermentation process of claim 1, wherein the cellulase mixture comprises cellulases and hemicellulases in a ratio of at least 2:

1.

4. The fermentation process of claim 1, wherein the cellulase mixture comprises cellulases and hemicellulases in a ratio of at least 3:

1.

5. The fermentation process of claim 1, wherein the carbon source comprises one or more additional carbon sources.

6. The fermentation process of claim 5, wherein the one or more additional carbon source is glycerol or an organic acid.

7. The fermentation process of claim 1, wherein the host cell is a strain of Trichoderma reesei.

8. The fermentation process of claim 1, wherein the process produces at least 2-fold more secreted cellulase when compared to the amount of secreted cellulose produced in a process in which the carbon source consists of hemicellulose-derived carbohydrates.

9. The fermentation process of claim 1, wherein the process is characterized by having at least a 3-fold increase in specific productivity (q_p) when compared to q_p in a process in which the carbon source consists of hemicellulose-derived carbohydrates.

10. The fermentation process of claim 1, wherein the process is characterized by having at least a 5-fold increase in specific productivity (q_p) when compared to q_p in a process in which the carbon source consists of hemicellulose-derived carbohydrates.

11. The fermentation process of claim 1, wherein the process is fed-batch.

12. The fermentation process of claim 1, wherein the process is continuous.

13. The fermentation process of claim 1, wherein the process is conducted aerobically.

14. A use of the cellulase mixture produced by the fermentation process of claim 1 for the hydrolysis of a cellulosic substrate.

15. A use of the cellulase mixture produced by the fermentation process of claim 1 for the hydrolysis of a pretreated lignocellulosic substrate.

16. A method of hydrolyzing a cellulosic substrate comprising,

adding the cellulase mixture produced by the fermentation process of claim 1 to the cellulosic substrate to produce a reaction mixture, wherein the concentration of cellulose in the reaction mixture is from about 1 g/L to about 200 g/L and the cellulase mixtures is added at a concentration from about 0.1 mg protein/g cellulose to about 100 mg protein/g cellulose;

incubating the reaction mixture for a period of time from about 4 to about 120 hours, at a temperature from about 30°

to about 60°

C., and at a pH from about 3.5 to about 7.0 to hydrolyze the cellulosic substrate and produce reaction products.

17. The method of claim 16, wherein the reaction products include hemicellulase-derived carbohydrate, glucose, oligosaccarides, cellulase-inducing carbohydrates, and a combination thereof.

18. The method of claim 16, wherein the cellulosic substrate is a pretreated lignocellulosic substrate.