Kasugamycin 생산을 위한 연속 청정생물공정 개발

김창준; 박선옥; 장용근; 전계택; 이종대; 김상용

Clean Technology, Vol.4, No.1, 45-59, June, 1998

Export Citation

Kasugamycin 생산을 위한 연속 청정생물공정 개발

Development of Continuous Clean Bioprocess for Kasugamycin Production

김창준, 박선옥, 장용근, 전계택, 이종대, 김상용

초록

방선균인 Streptomyces kasugaensis에 의해서 생산되는 이차대사산물인 kasugamycin 생산성 증대 및 단위 생산량 당 오염물질 배출량 저감을 위한 연속 고정화 배양을 수행하였다. 세포 고정화에 적합한 포자 수확을 위한 포자형성 촉진 배지 개발은 물론 고정화 담체로 사용한 셀라이트에 수확된 포자를 고정화시키는 방법을 확립하였다. 연속 고정화 배양 시 고정화세포의 유출을 방지함으로써 안정된 연속배양을 가능토록 하기 위한 decantor 형태의 고정화세포 분리기가 장착된 반응기를 사용하였다. 희석 속도 및 공급액 중의 기질 농도(당 농도, 인 농도)를 변화시키며 생합성된 kasugamycin 생산성 및 화학적산소요구량(COD)을 측정하였으며 이들을 현탁세포를 이용한 회분식 발효에서의 kasugamycin 생산성 및 발효 폐액 중의 COD 값과 비교하였다. 고정화 세포를 이용한 연속배양에서의 kasugamycin 생산성이 현탁세포를 이용한 회분식 발효에 비해 2.5배 높았으며, 동시에 단위 생산량 당 COD가 2.3배 저감되는 것으로 나타났다. 이것으로부터 연속 고정화 배양이 현탁 회분식 배양에 비해 kasugamycin 생산성 및 오염물질 배출 측면에서 유리한 청정 공정임을 확인할 수 있었다.

Continuous immobilized-cell culture was carried out for the production of kasugamycin, a secondary metabolite by a filamentous bacteria, Streptomyces kasugaensis, with an intention of reducing waste generation. A sporulation medium was developed for production of bulk amounts of spores, and the spores were entrapped into celite biosupports for immobilization. It was possible to effectively keep the immobilized-cells inside the reactor during the continuous culture by an efficient immobilized cell separator of decantor type on the outlet of the fermentor. Using this continuous immobilized-cell fermentor system, we investigated the effects of feed substrate and phosphate concentrations on kasugamycin production and chemical oxygen demand(COD). Comparing with the conventional suspended-cell batch culture, the kasugamycin productivity was observed to increase by 2.5 times, whereas COD per unit kasugamycin production decreased by 2.3 times in the continuous immobilized-cell culture. Based on these results, the continuous immobilized-cell system was considered to be a cleaner bioprocess than the conventional batch suspended-cell system.

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[1] Crueger W, Crueger A, "In Biotechnology, A Textbook of Industrial Microbiology," 2nd edn., Sinauer Associates, Inc., Sunderland, MA, pp. 229-273 (1989)

[2] Masurekar PS, "In Biotechnology of Filamentous Fungi," Butterworth-Heinemann, Boston, pp. 241-301 (1992)

[3] Stanbury PF, Whitakev A, Hall SJ, "Principles of Fernentation Technology," 2nd edn., Pergamon, London, pp. 13-33 (1995)

[4] Dalli M, Chau PC, Biotechnol. Lett., 10, 331 (1988)

[5] Berk D, Behie LA, Jones A, Lesser BH, Gaucher GM, Can. J. Chem. Eng., 62, 112 (1984)

[6] Jones A, Berk D, Lesser BH, Behie LA, Gaucher GM, Biotechnol. Lett., 5, 785 (1988)

[7] Oh DK, Hyun CK, Kim JH, Park YH, Biotechnol. Bioeng., 32, 569 (1988)

[8] Umezawa H, Okami Y, Hashimoto T, Suhara Y, Hamada M, Takenchi T, J. Antibiot. Ser. A, 18, 101 (1965)

[9] Fukagawa Y, Sawa T, Takeuchi T, Umezawa H, J. Antibiot., 21, 50 (1968)

[10] Ichikawa T, Date M, Ozaki A, Improvement of Kasugamycin-Producing Strain by The Agar Piece Method and the Prototroph Method, Folia Microbiologica, 16, 218 (1971)

[11] Yagi Y, Kitamura I, Okamura K, Ozaki A, J. Ferment. Technol., 49, 117 (1971)

[12] Umezawa H, Japan Patent, 58170494;07.10.83 (1984)

[13] Takeuchi T, Ishizuka M, Takayama H, Kureha K, Hamada M, Umezawa H, J. Antibiot. Ser. A, 18, 107 (1965)

[14] Hamada M, Hashimoto T, Takahashi T, Yokoyama S, Miyake M, Takeuchi T, Okami Y, Umezawa H, J. Antibiot. Ser. A., 18, 104 (1965)