Immobilized biocatalysts

• US 5,137,818 A
• Filed: 07/02/1990
• Issued: 08/11/1992
• Est. Priority Date: 11/15/1985
• Status: Expired due to Term

• Current Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Gist-Brocades N.V. (DSM-Firmenich AG)
• Inventors: Harder, Abraham, Cummings, Marsha, DeHaan, Ben R., Van Der Plaat, Johannes B.
• Primary Examiner(s): Lilling, Herbert J.
• Application Number:
  US07/547,868
• Time in Patent Office: 771 Days
• Field of Search: 435/174, 435/177, 435/179, 435/180, 435/182, 435/240.22
• US Class Current: 435/177
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 11/18 Multi-enzyme systems
• CPC Class Codes: C12P 7/06 Ethanol, i.e. non-beverage
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y10S 435/942 Saccharomyces cerevisiae

Abstract

Immobilized water-insoluble biocatalysts in particulate form comprise living cells, particularly yeast, dispersed in a cross-linked gelling agent. An enzyme, particularly amyloglucosidase, may be co-immobilized in the particles. These particles are prepared by suspending the living cells in an aqueous solution of a gelling agent, dispersing this suspension in a water immiscible organic liquid to form a suspension in the liquid of aqueous particles comprising the living cells and gelling agent, gelling the gel and cross-linking the gelling agent. It is found that when living cells such as microbial cells and especially yeast are immobilized in this way, that surprisingly, not only is their viability retained, but the ability of yeast cells to produce ethanol under continuous fermentation conditions is significantly improved. Specific strains of Saccharomyces cerevisiae, suitable for immobilization in this way, are described.

Claims

1. An immobilized water-insoluble biocatalyst in particulate form consisting essentially of living cells in a gelling agent comprising gelatin which is cross-linked with a cross-linking agent, the biocatalyst being further characterized in that at least part of the water is removed from the cross-linked particles to give a dry matter content of at least 15%.

2. A biocatalyst according to claim 1 wherein the living cells are selected from the group consisting of yeasts, bacteria and plant cells, or mixtures thereof.

3. A biocatalyst according to claim 1 wherein the living cells are dispersed in conjunction with one or more enzymes which are compatible with the living cells and the gelling agent.

4. A biocatalyst according to claim 3 wherein the enzymes are selected from the group consisting of amyloglucosidase, lactase, maltase, amylase, glucose isomerase, pullulanase, invertase, lipase, esterase, glucose oxidase, glucose dehydrogenase, cellulase, hemicellulase and xylanase.

5. A biocatalyst according to claim 1 wherein the living cells are an ethanol sensitive strain of S. cerevisiae having a D-value (the time required to kill 90% of the living cells at 58°

C.) of more than 25 seconds.

6. A biocatalyst according to claim 1 which comprises viable S. cerevisiae cells optionally in conjunction with amyloglucosidase dispersed in cross-linked gelatin.

7. A biocatalyst according to claim 1 wherein the living cells are a thermophilic strain of S. cerevisiae capable of sustained growth in yeast extract/peptone/glucose at a temperature in excess of 40°

C.

8. A biocatalyst according to claim 7 wherein the thermotolerant strain is S. cerevisiae CBS 252.86.

9. A biocatalyst according to claim 6 wherein the viable cells are of the strain S. cerevisiae CBS 253.86.