Production of cyclosporin A and/or C with a strain of NECTRIA

Details Production of cyclosporin A and/or C with a strain of NECTRIA Production of cyclosporin A and/or C with a strain of NECTRIA

1993-11-10

1995-09-05

Marx, Irene

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Using a micro-organism to make a protein or polypeptide

435 711, 4352541, C12P 2104, C12N 114

Patent

active

054478545

DESCRIPTION:

BRIEF SUMMARY
DESCRIPTION

1. Technical Field
This invention relates to a method of producing cyclosporin A and/or C each having immunosuppressant, antiinflammatory and other activities. The invention finds application in the manufacture of drugs, among other things.
2. Background Art
The structures of cyclosporin A and C are disclosed in Helvetica Chimica Acta 59, 1075-1092 (1976), ditto 60, 1247-1255 (1977) and ditto 70., 13-36 (1987) and the technology for producing cyclosporin compounds, inclusive of cyclosporin A and C, are described in JP Kokai S-50-89598, JP Kokai S-52-59180, JP Kokai S-55-55150 and JP Kokai S-57-63093.


DISCLOSURE OF INVENTION

The inventors of this invention explored for a new technology for producing cyclosporin compounds and discovered that a new strain of microorganism of the genus Nectria(Fr.)Fr., which differentiates itself from any of the organisms employed in the above-mentioned prior art technology, is able to produce cyclosporin A and/or C. This invention is, therefore, directed to a method which comprises culturing a cyclosporin A and/or C-producing strain of microorganism of the genus Nectria and recovering cyclosporin A and/or C from the resulting cultured broth.
Among the cyclosporin A and/or C-producing strains belonging to the genus Nectria which can be employed in this invention, the strain (designated F-4908) which the inventors of this invention isolated from a soil sample collected in NaZe-shi, Amami Ohshima, Kagoshima-ken, Japan has the following mycological characteristics.
Cultural characteristics on various media
Table 1 shows cultural characteristics of F-4908 on potato glucose agar, malt extract agar and corn meal agar after 2 weeks incubation at 25.degree. C. The color descriptions used in this specification are based on the Color Standard issued by Japan Color Research Institute.


TABLE 1 ______________________________________ Cultural characteristics of F-4908 on various media Potato glucose Malt extract Corn meal agar agar agar ______________________________________ Colony morphology irregular irregular to Circular circular Colony diameter 1.5 cm 1.5-2.0 cm 2.5-3.0 cm Colony surface Wrinkled and Wrinkled in Plane and raised, felt- center, plane, thin, aerial like and felt-like and mycelium covered with covered with not rising. short flocci. short flocci. Surface color Light olive - Pale olive - White. olive or light olive. Yellow in grayish olive. center Orange-colored only. exudations pro- duced at center. Reverse color Dark yellowish Yellowish White. brown-dark brown. A Yellow in brown. A yellow solu- center brown soluble ble pigment only. pigment diffus- diffusing ing into med- into medium. ium. Other No spore No spore Conidia are characteristics formation in formation in formed on teleomorphic teleomorphic 2-week and anamorphic and anamor- incubation states. phic states. at 25.degree. C.. Perithecia are formed on 4-week or longer incubation at 25.degree. C.. ______________________________________
Physiological characteristics
The growth temperature range of F-4908 is 4-35.degree. C. and the optimum temperature range for growth is 18-28.degree. C. The growth pH range of the strain is pH 3-10 and the optimum pH range is pH 6-7.
The teleomorph(ascomata) of F-4908 is observed on corn meal agar at 25.degree. C. after not less than 4 weeks, with the production of brown ascospores in the perithecia. The hyphal conidiomata is observed on various culture media. The conidiogenesis is enteroblastic (phialidic).
When the stain is inoculated on a plant leaf whose surface has been sterilized, the perithecia are superficial and globose, subglobose or ampule-shaped, with one papillate ostiole. There is no lateral hair, the color ranges from light brown to orange, and the diameter is 150-250 .mu.m. The peridia are composed of 3 to 4 layers of thin-walled cells. The papillae are 70 to 80 .mu.m in diameter and 50-60 .mu.m high, and there a

REFERENCES:
patent: 4108985 (1978-08-01), Ruegger et al.
patent: 4330560 (1982-05-01), Staron et al.
patent: 5051402 (1991-09-01), Kurihara et al.
patent: 5156960 (1992-10-01), Jekkel
patent: 5214130 (1993-05-01), Patchett et al.
patent: 5256547 (1993-10-01), Rudat et al.
ATCC Catalogue of Funji/Yeohi, 17th Ed, 1987, pp. 227-229.

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