# US 5,344,765 A · DNA encoding polypeptides with activity against gram-positive and gram-negative bacteria

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- Number: US 5,344,765 A
- Document ID: US5344765A
- Status: Granted
- Kind: A
- Application number: 08/010,676
- Filed: 1993-01-28
- Granted: 1994-09-06
- Earliest claimed: 1985-01-28
- Assignees: Xoma Corporation
- Inventors: Jiunu S. Lai; Jar-How Lee; James E. Callaway
- Web page: https://patentgrep.com/patent/US5344765A

## CPC

- C07K 14/52: Cytokines; Lymphokines; Interferons
- C07K 14/43563: {from insects}
- C07K 14/585: Calcitonins
- C12N 15/62: DNA sequences coding for fusion proteins
- C12N 15/70: Vectors or expression systems specially adapted for E. coli
- C07K 2319/00: Fusion polypeptide
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention relates to cecropin-like polypeptides with a variety of C-terminus modifications. Conversion of the terminal carboxy group to a homoserine extended neutral, negatively charged or positively charged form results in the polypeptides of the invention that have significant bactericidal effects against both Gram-positive and Gram-negative bacteria.

## Claims

### Claim 1

A recombinant DNA molecule comprising a genetic sequence coding for a cecropin polypeptide which is extended at the C-terminus thereof by Met-X wherein X is a group which is clearable from methionine by CNBr.

### Claim 2 (depends on 1)

The recombinant DNA molecule of claim 1, wherein X is an amine acid or peptide residue.

### Claim 3 (depends on 1)

The recombinant DNA molecule of claim 1, wherein said cecropin polypeptide is homologous to cecropin A.

### Claim 4 (depends on 2)

The recombinant DNA molecule of claim 2, wherein said cecropin polypeptide is homologous to cecropin A.

### Claim 5 (depends on 1, 2, 3, 4)

An expression vehicle replicable in a given host comprising the recombinant DNA molecule of any of claims 1-4.

### Claim 6 (depends on 5)

The expression vehicle of claim 5, which is a plasmid.

### Claim 7 (depends on 5)

The expression vehicle of claim 5, wherein said host is a bacterium.

### Claim 8 (depends on 5)

A method of producing a cecropin polypeptide which is extended by a homoserine residue at the C-terminus thereof which comprises:
transforming a suitable host with the expression vehicle of claim 5;
culturing said host;
recovering said cecropin polypeptide, having a Met-X at the C-terminus thereof; and
cleaving said Met-X to thereby produce a C-terminal homoserine.

### Claim 9

A pharmaceutical composition with antimicrobial activity, comprising an effective amount of a cecropin polypeptide which is extended by a homoserine residue at the C-terminus thereof.