# US 5,703,049 A · High methionine derivatives of .alpha.-hordothionin for pathogen-control

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- Number: US 5,703,049 A
- Document ID: US5703049A
- Status: Granted
- Kind: A
- Application number: 08/608,786
- Filed: 1996-02-29
- Granted: 1997-12-30
- Earliest claimed: 1996-02-29
- Assignees: Pioneer Hi-Bred Int'l., Inc.
- Inventors: Aragula Gururaj Rao
- Web page: https://patentgrep.com/patent/US5703049A

## CPC

- C07K 14/415: from plants
- C12N 15/8282: {for fungal resistance}
- 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

Derivatives of .alpha.-hordothionin made by position-specific substitution with methionine residues provide methionine enrichment in plants while retaining the anti-pathogenic activity of the parent compound.

## Claims

### Claim 1

A method for killing and inhibiting plant pathogenic microorganisms which are susceptible to a .alpha.-Hordothionin comprising introducing into the environment of the pathogenic microorganisms an anti-microbial amount of a protein having the sequence of SEQUENCE I.D. NO. 3 wherein the amino acid residues at one or more of positions 1,5,8,11,15,17,18,19,22,23,24,30,32, 33,38, and 41 are methionine, and the remainder of the residues at those positions are the residues at the corresponding positions in SEQUENCE I.D. NO. 1.

### Claim 2 (depends on 1)

The method of claim 1 wherein one or more of the amino acid residues at positions 8,11,15,18,22,33, and 41 are methionine.

### Claim 3 (depends on 2)

The method of claim 2 wherein at least three of the amino acid residues at positions 8,11,15,18,22,33, and 41 are methionine.

### Claim 4 (depends on 3)

The method of claim 3 wherein at least five of the amino acid residues at positions 8,11,15,18,22,33, and 41 are methionine.

### Claim 5 (depends on 4)

The method of claim 4 wherein the environment of the pathogen is the tissues of a living plant.

### Claim 6

A method for killing and inhibiting plant pathogens selected from Fusarinm graminearum, Fusarinm moniliforme, Diplodia maydis, Colletototrichnm graminicola, Verticillium alboatrum, Phytophthora megaspermae f.sp. glycinea, Macrophomina phaseolina, Diaporthe phaseolorum cavlivora, Sclerotinia sclerotiornm, Sclerotinia trifoliorum, and Aspergillus flavus, comprising introducing into the environment of the pathogenic microorganisms an anti-microbial amount of a protein having the sequence of SEQUENCE I.D. NO. 3 wherein the amino acid residues at one or more of positions 1,5,8, 11,15,17,18,19,22,23,24,30,32,33,38, and 41 are methionine, and the remainder of the residues at those positions are the residues at the corresponding positions in SEQUENCE I.D. NO. 1.

### Claim 7 (depends on 6)

The method of claim 6 wherein the environment of the pathogen is the tissues of a living plant.

### Claim 8 (depends on 7)

The method of claim 7 wherein one or more of the amino acid residues at positions 8,11,15,18,22,33, and 41 are methionine.

### Claim 9 (depends on 8)

The method of claim 8 wherein at least three of the amino acid residues at positions 8,11,15,18,22,33, and 41 are methionine.

### Claim 10 (depends on 9)

The method of claim 9 wherein at least five of the amino acid residues at positions 8,11,15,18,22,33, and 41 are methionine.