Rapamycin producer

Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor

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Details

435119, 435 721, 435117, 435111, C12N 120

Patent

active

056747322

DESCRIPTION:

BRIEF SUMMARY
THIS APPLICATION WAS FILED UNDER 35 USC 371 AS THE NATIONAL PHASE OF
PCT/US93/01534.


TECHNICAL FIELD

This invention relates to a new culture capable of producing rapamycin. Rapamycin of the present invention is produced by fermentation of a new microorganism designated as Actinoplanes sp. N902-109 (FERM BP-3832) in a nutritious medium.


BACKGROUND ART

Rapamycin (U.S. Pat. No. 3929992 and No. 3993749) was reported as an antifungal antibiotic which was produced by Streptomyces hygroscopicus AY B-994 (ATCC 29253) (C. Vezina, A. Kudelski and S. N. Sehgal, J. Antibiotics 28,721-726, 1975). In recent years, it has been demonstrated that rapamycin shows potent immunosuppressive activity. Hitherto, it is known that S. hygroscopicus ATCC 29253 is used for the production of rapamycin, which was reported together with physicochemical properties (U.S. Pat. No. 3929992). An object of the present invention is to provide a new culture which belongs to the different genus from Streptomyces. In addition, an object of the present invention is to provide a culture capable of producing rapamycin more efficiently than the cultures which have been reported in the prior art.


BRIEF DISCLOSURE OF THE INVENTION

The present invention is directed to a new culture capable of producing rapamycin. Rapamycin is produced by fermentation of Actinoplanes sp. N902-109 (FERM BP-3832).
The present invention also is directed to a culture capable of producing rapamycin more efficiently than Streptomyces hygroscopicus which has been reported in the prior art.
Furthermore, the present invention is directed to a biologically-pure culture having the characteristics of Actinoplanes sp. N902-109 (FERM BP-3832), as well as mutants and transformants of any of the foregoing, capable of producing rapamycin, including any such culture in freeze-dried form. Such a culture is capable of producing rapamycin in a recoverable quantity upon fermentation in an aqueous nutrient medium comprising assimilable sources of carbon and nitrogen.


DETAILED DESCRIPTION OF THE INVENTION

A culture N902-109 was isolated from a soil sample collected in Shizuoka Prefecture, Japan. The culture has narrow hyphae of the Actinomycetales and motile spores produced in sporangia-like clusters on the substrate mycelium--a feature characteristic of members of the genus Actinoplanes. The generic identity was further supported by the results of whole-cell analysis.
The taxonomical properties of N902-109 are as follows. The culture N902-109 was planted from a slant into ATCC #172 broth and grown for 4 days at 28.degree. C. on a shaker. It was then centrifuged for 20 minutes, washed 3 times with sterile water and planted on media commonly used for identification of members of the Actinomycetales. The culture was incubated at 28.degree. C. and the results were read at varying times but most commonly were taken at 14 days. The colors were described in common terminology, but exact colors were determined by comparisons with color chips from Color Harmony Manual, 4th Ed. The methods of whole-cell amino acid and sugar analyses are those described in Becker, B. et al., Appl. Microbiol., 12, 421-423, 1964, and in Lechevalier, M. P., J. Lab. Clin. Med., 71, 934-944, 1968.
Identification media used for the characterization of the culture and references for their composition are as follows: no. 1, p. 328, 1961. Bacteriol., 69, 147-150, 1955. medium no. 1, p. 328, 1961, with 3 g dextrose substituted for 30 g sucrose and agar omitted. 934-944, 1968, but use only 30 g potatoes, 2.5 g carrots and 20 g agar. Classification of Antagonistic Actinomycetes, English Ed., p.13, 1957. 231-240, 1977. 231-248, 1930. b) M. Levine and H. W. Schoenlein, A Compilation of Culture Media, medium no. 2511, 1930. Brodsky, Antimicrob. Agents and Chemother., 1964, 47-52, 1965.
Yeast Extract-Malt Extract Agar--Growth good, orange (4 la, 4 lc); raised, wrinkled, no aerial mycelium; reverse orange (4 la, 4 na); soluble pigment yellowish brown (3 lc).
Oatmeal Agar--Growth moderate, pale orange yellow (3 ea, 4 ea); sli

REFERENCES:
Rose, A. H. "Industrial/Microbiology", Butter-Worths, London, 1961, pp. 191-193.

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