Process for producing cellulolytic enzymes

• US 4,762,788 A
• Filed: 11/28/1984
• Issued: 08/09/1988
• Est. Priority Date: 11/29/1983
• Status: Expired due to Term

• Current Assignee: Institut Francais Du Petrole
• Original Assignee: Institut Francais Du Petrole
• Inventors: Ferre, Veronique, Pourquie, Jacques, Warzywoda, Michel
• Primary Examiner(s): Warren, Charles F.
• Assistant Examiner(s): Hanley, Elizabeth A.
• Application Number:
  US06/675,595
• Time in Patent Office: 1,350 Days
• Field of Search: 435/209, 435/254, 435/945
• US Class Current: 435/71.1
• 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: Y10S 435/945 Trichoderma

Abstract

This invention concerns a process of producing cellulolytic enzymes by fermentation with the Trichoderma reesei fungus.

This process includes two steps:

(a) a first step of subjecting to aerobian fermentation a culture medium comprising a strain of Trichoderma reesei, nutrition compounds and a small quantity of cellulose and sugar,

(b) a second step of contining the aerobian fermentation by adding soluble sugar at a speed such that the sugar concentration in the fermentation zone remains under 0.3% by weight.

The cellulolytic enzymes obtained are used in the enzymatic hydrolysis of a lignocellulosic biomass.

Claims

1. A process for aerobic production of cellulase by cultivation of a Trichoderma reesei strain in a fermentation zone containing a nutrient medium, said process comprising feeding said fermentation zone with soluble sugar and being characterized by:

(a) a first step of subjecting to aerobic fermentation conditions a culture medium comprising a Trichoderma reesei strain, an inorganic salt and an initial carbon substrate containing at least one cellulose containing material and at least one soluble sugar, the amount by weight of said cellulose and said soluble sugar, in proportion to the culture medium, being each 0.01-2%, the first step being continued without further soluble sugar addition for a sufficient time to consume at least 10% of the sugar and to obtain a sugar concentration in the medium lower than 0.3% by weight, and

(b) a second step of continuing the aerobic fermentation by adding soluble sugar continuously so as to introduce a sugar amount totalling at least 3% by weight of the culture medium while maintaining the sugar concentration in said culture medium at a value lower than 0.3% by weight.

2. A process according to claim 1, wherein no cellulose-containing material is added during the second step.

3. A process according to claim 1, wherein the soluble sugar is lactose, glucose, xylose, arabinose, cellobiose or a mixture thereof.

4. A process according to claim 1, wherein the fermentation zone is fed with soluble sugar during the second step in a continuous manner.

5. A process according to claim 1, wherein the cellulose-containing material is paper pulp.

6. A process according to claim 1, wherein in step (a) the cellulose and sugar amounts are respectively 0.2-2% and 0.02-1% by weight and the contact is maintained until the sugar concentration becomes lower than 0.2% by weight and at least 20% of the sugar has been consumed.

7. A process according to claim 1, wherein the sugar 30 amount in step (a) is 10-50% by weight of the cellulose amount and the sugar concentration during step (b) is 0.1-0.25% by weight.

8. A process according to claim 3, wherein the soluble sugar is lactose.

9. A process according to claim 1, wherein the concentration of sugar in step (b) is maintained at a value of 0.1 to 0.3% by weight.

10. A process according to claim 1, wherein a hyperproducing Trichoderma reesei strain is used.