# US 4,397,947 A · Microbial process for 9.alpha.-hydroxylation of steroids

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 4,397,947 A
- Document ID: US4397947A
- Status: Granted
- Kind: A
- Application number: 06/273,145
- Filed: 1981-06-12
- Granted: 1983-08-09
- Earliest claimed: 1981-06-12
- Assignees: G. D. Searle & Co.
- Inventors: William J. Marsheck; James Jiu; Ping T. Wang
- Web page: https://patentgrep.com/patent/US4397947A

## CPC

- C12P 33/06: Hydroxylating
- Y10S 435/872: Nocardia
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Valuable 9.alpha.-hydroxy steroids are prepared via microbial enzymatic oxidation (MEO) by conventional fermentation means, without the need for a .DELTA..sup.1 dehydrogenation inhibitor, utilizing novel microorganism Nocardia canicruria ATCC 31548. Also disclosed is a novel method using any .DELTA..sup.1 dehydrogenase producing organism and a novel bio-reactor technique means for preparing said steroids.

## Claims

### Claim 1

A process for 9.alpha.-hydroxylating steroids which comprises subjecting a 9-unsubstituted steroid to the microbial enzymatic oxidation action of Nocardia canicruria ATCC 31548, by fermentation means.

### Claim 2 (depends on 1)

A process according to claim 1 wherein said fermentation means are the conditions necessary for the growth of Nocardia canicruria ATCC 31548.

### Claim 3 (depends on 2)

A process according to claim 2 wherein said 9-unsubstituted steroid is from the gonane series.

### Claim 4 (depends on 2)

A process according to claim 2 wherein said 9-unsubstituted steroid is from the estrane series.

### Claim 5 (depends on 2)

A process according to claim 2 wherein said 9-unsubstituted steroid is from the androstane series.

### Claim 6 (depends on 2)

A process according to claim 2 wherein said 9-unsubstituted steroid is from the pregnane series.

### Claim 7 (depends on 2)

A process according to claim 2 wherein said 9-unsubstituted steroid is from the cholestane series.

### Claim 8

A bio-reactor process for 9.alpha.-hydroxylating steroids which comprises subjecting a 9-unsubstituted steroid which is dissolved in a chlorinated hydrocarbon to the enzymatic action of a biomass of Nocardia canicruria ATCC 31548 in the presence of a source of air and at elevated temperature.

### Claim 9 (depends on 8)

A process according to claim 8 wherein said chlorinated hydrocarbon is carbon tetrachloride.

### Claim 10 (depends on 9)

A process according to claim 9 wherein said 9-unsubstituted steroid is ethisterone acetate.

### Claim 11 (depends on 9)

A process according to claim 9 wherein said 9-unsubstituted steroid is methyl 3-oxopregna-4,17(20)-dione-20-carboxylate.

### Claim 12

A biologically pure culture of the microorganism Nocardia canicruria ATCC 31548, wherein said culture is capable of producing 9.alpha.-hydroxyl steroids from 9-unsubstituted steroids by fermentation means in recoverable amounts.