# US 5,683,913 A · Regulatory gene for the expression of nitrile hydratase gene

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- Number: US 5,683,913 A
- Document ID: US5683913A
- Status: Granted
- Kind: A
- Application number: 08/537,434
- Filed: 1995-10-02
- Granted: 1997-11-04
- Earliest claimed: 1994-10-03
- Assignees: Sakayu Shimizu; Nitto Chemical Industry Co., Ltd.
- Inventors: Sakayu Shimizu; Michihiko Kobayashi
- Web page: https://patentgrep.com/patent/US5683913A

## CPC

- C12N 9/78: acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C07K 14/195: from bacteria
- C12N 15/74: Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The invention relates to a regulatory gene coding for a polypeptide having the ability to activate a promoter for a nitrile hydratase gene, a recombinant DNA containing said regulatory gene, and a transformant transformed with said recombinant DNA. The introduction of the regulatory gene of the invention along with a nitrile hydratase gene and its promoter region permits bacteria of the genus Rhodococcus to produce a higher level of nitrile hydratase. Other extraneous gene can also be introduced into a region downstream of the promoter to produce other proteins in high yield.

## Claims

### Claim 1

An isolated regulatory gene coding for a polypeptide having the ability to activate a promoter for a nitrile hydratase gene and having the amino acid sequence defined in the Sequence Listing by SEQ ID No: 1.

### Claim 2 (depends on 1)

The gene according to claim 1 wherein the regulatory gene coding for a polypeptide having the ability to activate a promoter for a nitrile hydratase gene possesses the nucleotide sequence defined in the Sequence Listing by SEQ ID No: 2.

### Claim 3 (depends on 1, 2)

A recombinant DNA wherein the regulatory gene of claim 1 or 2 and a nitrile hydratase gene containing a promoter region have been inserted into a vector.

### Claim 4 (depends on 3)

A transformed host cell containing the recombinant DNA of claim 3.