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Tolypocladium inflatum을 이용한 Cyclosporin A 발효에서 아미노산과 유기질소원의 영향

Effect of Amino Acids and Organic Nitrogen Sources on Cyclosporin A Fermentation by Tolypocladium inflatum

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초록

Cyclosporin, an immunosuppressant, is a representative group of biologically active secondary metabolites produced by the fungus Tolypocladium inflatum. The amount and ratio of cyclosporin derivatives in the culture broth are an important factors for the production of cyclosporin A and the purification in the industrial process. Therefore, we studied the effect of amino acids and complex organic nitrogen sources using Tolypocladium inflatum mutants on the productivity of cyclosporin A and the ratio of cyclosporin derivatives. Overproducing mutant YHC-004 having seven times higher productivity than mother strain's could be obtained through the artificial mutation by UV irradiation. The concentration and kind of organic nitrogens and amino acids shows the profound effect on the productivity of cyclosporin A and ratio of cyclosporin derivatives. As a result, it was possible to raise the productivity and the ratio of cyclosporin A up to 3,430 mg/L and 93% respectively, but on the other hand the other cyclosporin derivatives decreased less than 2% in the culture broth.

Abstract

Cyclosporin, an immunosuppressant, is a representative group of biologically active secondary metabolites produced by the fungus Tolypocladium inflatum. The amount and ratio of cyclosporin derivatives in the culture broth are an important factors for the production of cyclosporin A and the purification in the industrial process. Therefore, we studied the effect of amino acids and complex organic nitrogen sources using Tolypocladium inflatum mutants on the productivity of cyclosporin A and the ratio of cyclosporin derivatives. Overproducing mutant YHC-004 having seven times higher productivity than mother strain's could be obtained through the artificial mutation by UV irradiation. The concentration and kind of organic nitrogens and amino acids shows the profound effect on the productivity of cyclosporin A and ratio of cyclosporin derivatives. As a result, it was possible to raise the productivity and the ratio of cyclosporin A up to 3,430 mg/L and 93% respectively, but on the other hand the other cyclosporin derivatives decreased less than 2% in the culture broth.

참고문헌 (20)

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  2. Agathos, S. N, "Physiological and genetic factors for process development of cyclosporine fermentations", J. Ind. Microbiol, 1, pp.39~47 (1986)
  3. Agathos, S. N, "The fungal production of cyclosporin", Ann. N.Y. Acad. Sci, 506, pp.657~662 (1987)
  4. Billich, A, "Enzymatic synthesis of cyclosporin A", J. Biol. Chem, 262, pp.17258~17259 (1987)
  5. Borel, J. F, "Cyclosporin and its future. in: Cyclosporins, Progress in allergy, vol. 38" (1986), Karger
  6. Chun, G-T, "Comparative studies of physiological and environmental effects on the production of cyclosporin A in suspended and immobilized cells of Tolypocladium inflatum", Biotechnol. Bioeng, 37, pp.256~265 (1991)
  7. Dreyfuss, M, "Cyclosporin A and C - New metabolites from Trichoderma polysporum", Eur. J. Appl. Microbiol, 3, pp.125~133 (1976)
  8. El Enshasy, H, "Kinetics of Cell Growth and Cyclosporin A Production by Tolypocladiuminflatum when Scaling Up from Shake Flask to Bioreactor", Journal of Microbiology and Biotechnology, 18(1), pp.128~134 (2008), 한국미생물·생명공학회
  9. Fang, J. R, "The effect on the biosynthesis of cyclosporin A by the addition of amino acids", Clin. J. Antibiot, 15, pp.140~141 (1990)
  10. Issac, C. E, "The production of cyclosporins by Tolypocladium niveum strains. Antimicrob", Agents Chemother, 34, pp.121~127 (1990)
  11. Kobel, H, "Directed biosynthesis of cyclosporins", Eur. J. Appl. Microbiol. Biotechnol., 14, pp.237~240 (1982)
  12. Kreuzig, F, "High speed liquid chromatography with conventional instruments for the determination of cyclosporin A, B, C, and D in fermentation broth", J. Chromatogra, 290, pp.181~186 (1984)
  13. Lee, J, "Effect of amino acids on the production of cyclosporin A by Tolypocladium inflatum", Biotechnol. Lett, 11, pp.77~82 (1989)
  14. Lee, J, "Dynamics of L-valine in relation to the production of cyclosporin A by Tolypocladium inflatum", Appl. Microbiol. Biotechnol., 34, pp.513~517 (1991)
  15. Lee, Mi-Jin, "Spore Inoculum Optimization to Maximize Cyclosporin A Production inTolypocladium niveum", Journal of Microbiology and Biotechnology, 18(5), pp.913~917 (2008), 한국미생물·생명공학회
  16. Margaritis, A, "Development of a fructose based medium for biosynthesis of cyclosporin A by Beauveria nivea", Biotechnol. Lett, 11, pp.765~768 (1989)
  17. Sekar, C, "Optimization studies on the production of ctcosporin A by solid state fermentation", Bioprocess Eng, 18, pp.293~296 (1998)
  18. Traber, R, "Cyclosporins - new analogs by precursor directed biosynthesis", J. Antibiot, 42, pp.591~596 (1988)
  19. Wenger, R. M, "Synthesis of cyclosporin", Helv. Chim. Acta, 67, pp.502~506 (1984)
  20. Zocher, R, "Biosynthesis of cyclosporin A", Phytochemistry, 23, pp.549~551 (1984)

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