# US 9,492,500 B2 · Microcin and uses thereof

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- Number: US 9,492,500 B2
- Document ID: US9492500B2
- Status: Granted
- Kind: B2
- Application number: 14/407,975
- Filed: 2013-06-14
- Published: 2015-06-18
- Granted: 2016-11-15
- Earliest claimed: 2012-06-15
- Assignees: Washington State University
- Applicants: Washington State University
- Inventors: Douglas R. Call; Lauren Eberhart; Kelly A. Brayton; Thomas E. Besser; Ashish Sawant; Lisa Orfe
- Web page: https://patentgrep.com/patent/US9492500B2

## CPC

- A61K 38/164: {from bacteria}
- A01N 63/50: Isolated enzymes; Isolated proteins (peptides A01N37/46)
- A61K 35/74: Bacteria (therapeutic use of a bacterial protein A61K38/00)
- A61P 31/04: Antibacterial agents
- C07K 14/245: Escherichia (G)
- C11D 3/381: {Microorganisms}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12N 2710/10021: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2710/14021: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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)
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Microcin MccPDI and bacteria harboring the mcpM gene which encodes MccPDI, limit growth of and/or kill pathogenic bacteria such as pathogenic Escherichia coli (E. coli) and/or Shigella bacteria via proximity-dependent inhibition (PDl).

## Claims

### Claim 1

A method of killing or decreasing adverse effects of pathogenic Escherichia coli (E. coli) and/or Shigella bacteria, comprising
identifying a surface or subject known to or suspected of contamination with said pathogenic E. coli and/or Shigella bacteria; and
contacting said pathogenic E. coli and/or Shigella bacteria with
microcin MccPDI having SEQ ID NO: 24 or a functional variant thereof, or
an organism that produces MccPDI SEQ ID NO: 24 or a functional variant thereof,
wherein said functional variant has a sequence at least 95% identical to SEQ ID NO: 24.

### Claim 2 (depends on 1)

The method of claim 1, wherein said pathogenic Escherichia coli bacteria are Shiga-toxin E. coli (STEC) bacteria.

### Claim 3 (depends on 1)

The method of claim 1, wherein said STEC bacteria are selected from the group consisting of: enterohaemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enteroggregative E. coli (EAEC), diffusively adherent E. coli (DAEC), uropathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC).

### Claim 4 (depends on 2)

The method of claim 2, wherein said STEC bacteria are of a serotype selected from the group consisting of O111, O145, O103, O26, O45, O121 and O157.

### Claim 5 (depends on 1)

The method of claim 1, wherein said pathogenic Shigella bacteria are Shigella flexneri, Shigella sonnei or Shigella dysenteriae.

### Claim 6 (depends on 1)

The method of claim 1, wherein said organism that produces microcin MccPDI is a naturally occurring bacteria.

### Claim 7 (depends on 6)

The method of claim 6, wherein said naturally occurring bacteria are E-25 or E-264.

### Claim 8 (depends on 1)

The method of claim 1 wherein said organism that produces microcin MccPDI is a genetically modified organism harboring a heterologous nucleic acid which is expressed to produce said microcin MccPDI having SEQ ID NO. 24 or said functional variants thereof.