# US 4,393,047 A · Antibiotic cytophagin and a process for producing the same

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- Number: US 4,393,047 A
- Document ID: US4393047A
- Status: Granted
- Kind: A
- Application number: 06/279,489
- Filed: 1981-07-01
- Granted: 1983-07-12
- Earliest claimed: 1980-03-18
- Assignees: Microbiochemical Research Foundation
- Inventors: Hamao Umezawa; Tomio Takeuchi; Hiroshi Nakano
- Web page: https://patentgrep.com/patent/US4393047A

## CPC

- C12P 1/04: by using bacteria
- C12N 1/205: Bacterial isolates
- C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales}
- Y10S 435/822: using bacteria or actinomycetales
- 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)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

A substance having antibiotic activity, designated as cytophagin, which is stable in the form of colorless powder and which has an elemental analysis of H: 6.94%, C: 47.22% and N: 13.21%, a molecular weight of 1,000 to 1500. This substance is believed to be composed of 11 amino acids and exhibits a strong activity against Gram-positive bacteria. Cytophagin is produced by fermentation using a microorganism belonging to the genus Cytophaga and capable of producing cytophagin. A preferred strain is Cytophaga BMF 694-N3 (FERM P-4846;NRRL B-12109). Cytophagin is of potential interest as medicament or veterinary agent because of its antibiotic activity.

## Claims

### Claim 1

A substance having antibiotic activity, designated as cytophagin, which is stable in the form of colorless powder and which possesses the following physico-chemical properties:
(1) Elemental analysis: H: 6.94%, C: 47.22%, N: 13.21%
(2) Molecular weight: 1,000 to 1,500 [determined by gel filtration]
(3) Melting point: Browning at about 210.degree. C. and decomposing with foaming at about 220.degree. C.
(4) Specific rotation: [.alpha]sub.D.sup.25 =+5.degree. (c=1 in DMSO)
(5) Ultraviolet absorption spectrum: As shown in FIG. 1 (c=25 .mu.g/ml in methanol)
(6) Infrared absorption spectrum: As shown in FIG. 2 (by KBr tablet method)
(7) Solubility invarious solvents: Readily soluble in acetic acid and dimethylsulfoxide, substantially soluble in methanol and ethanol, and hardly soluble in water, chlorofoem, acetone, ethyl acetate and ethyl ether
(8) Color reaction: Positive in ninhydrin reaction, Ehrlich reaction and isatine reaction, and negative in Sakaguchi reaction, Pauly reaction and anisidine reaction.
(9) Nature: Neutral
(10) Color and appearance: Colorless powder
(11) Amino acids constituents: Lysine, aspartic acid, threonine, serine, proline, glycine, leucine, trypsine and other 3 amino acids (each one molecule thereof)
(12) Rf value: 0.23 [by silica gel thin layer chromatography using as silica gel plate Art. 5721 (Merck AG., Germany) and a solvent system of n-butanol/acetic acid/water (12:3:5)].

### Claim 2 (depends on 1)

The substance of claim 1, in which the molecular weight is 1,380 [calculated from the OD.sub.280 value of trypsine].

### Claim 3 (depends on 1)

A process for producing the substance claimed in claim 1, which process comprises the steps of culturing a microorganism selected from Cytophaga BMF 694-N3 (FERM P-4846; NRRL B-12109) and mutants thereof capable of producing the said substance in a culture medium under aerobic conditions to accumulate the said substance in the cultured broths and recovering the said substance therefrom.

### Claim 4 (depends on 3)

The process of claim 3, in which the culturing is effected at a temperature of from 10.degree. to 35.degree. C. for 2 to 5 days.