# US 4,399,222 A · Carbohydrases from acidophilic streptomyces

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- Number: US 4,399,222 A
- Document ID: US4399222A
- Status: Granted
- Kind: A
- Application number: 06/064,321
- Filed: 1979-08-06
- Granted: 1983-08-16
- Earliest claimed: 1979-08-06
- Assignees: A. E. Staley Manufacturing Company
- Inventors: Song H. Bok; LeRoy E. Jackson; Cynthia J. Schroedel; Martin Seidman
- Web page: https://patentgrep.com/patent/US4399222A

## CPC

- C12N 9/92: Glucose isomerase {(5.3.1.5; 5.3.1.9; 5.3.1.18)}
- Y10S 435/886: Streptomyces
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Acidophilic and acidoduric streptomycetes strains have been found to produce carbohydrases. These Streptomyces effectively elaborate glucose isomerase under acid conditions typically unfavorable for growth of conventional glucose isomerase producing Streptomyces. Sterilization of the culture and production media may be avoided by selectively propagating newly discovered Streptomyces acidodurans under acidic conditions which will effectively eliminate contaminating microorganisms. The Streptomyces acidodurans herein also have the ability to undergo cultivation and elaborate glucose isomerase over a relatively broad pH range. Constitutive streptomycetes strains have also been isolated. Glucose isomerases derived from these Streptomyces strains are particularly effective for isomerizing glucose syrups to fructose-containing syrups.

## Claims

### Claim 1

A method for producing glucose isomerase which comprises:
(a) inoculating and cultivating a culture medium containing assimilable carbon and nitrogen sources with microorganisms of the genus Streptomyces which will undergo cultivation at a pH less than 5.5, and produce glucose isomerase;
(b) continuing the cultivation for a period of time and under conditions sufficient to permit the microorganisms to produce glucose isomerase; and
(c) harvesting the glucose isomerase produced by said microorganisms from said culture medium.

### Claim 2 (depends on 1)

The method according to claim 1 wherein the Streptomyces is characterized as yielding at least one gram of Streptomyces dry cell per liter of culture medium and at least 10 glucose isomerase units/gm dry cells when cultivated at a pH 5.2.

### Claim 3 (depends on 1)

The method according to claim 1 wherein the microorganism consists generally of Streptomyces acidodurans or mutants thereof.

### Claim 4 (depends on 3)

The method according to claim 3 wherein the S. acidodurans is characterized as yielding at least 3 grams of dry cells/liter of culture medium and at least 150 G.I.U./gram of dry cells.

### Claim 5 (depends on 4)

The method according to claim 4 wherein the S. acidodurans has the ability to undergo cultivation and produce glucose isomerase at a pH 5.2 or less.

### Claim 6 (depends on 5)

The method according to claim 5 wherein the culture medium contains corn steep liquor substantially free from base-insoluble constituants in an amount sufficient to increase the glucose isomerase yield of said S. acidodurans.

### Claim 7 (depends on 4)

The method according to claim 4 wherein the S. acidodurans comprises a constitutive strain.

### Claim 8 (depends on 4)

The method according to claim 4 wherein the S. acidodurans is cultivated at a pH between about 3.5 to 5.0.

### Claim 9 (depends on 1)

The method according to claim 1 wherein the microorganism is at least one strain selected from the group consisting of S. acidodurans NRRL 11489, NRRL 11490, NRRL 11491, NRRL 11492, NRRL 11493, NRRL 11494, NRRL 11495, NRRL 11496, NRRL 11497, NRRL 11498 and mutants thereof.

### Claim 10 (depends on 1)

The method according to claim 1 wherein the recovered glucose isomerase is immobilized with a suitable carrier therefore.

### Claim 11 (depends on 5)

The method according to claim 5 wherein the glucose isomerase is immobilized with a suitable carrier therefore.