Development of a Culture Medium for Growth and Sporulation of Bacillus polyfermenticus SCD

프로바이오틱 비스루트균의 아포생산을 위한 최적배지 개발

  • 이광호 (경남대학교 생명과학부) ;
  • 박규용 (경남대학교 생명과학부) ;
  • 김성미 (경남대학교 생명과학부) ;
  • 김원석 ((주)바이넥스 기술연구소) ;
  • 백현동 (경남대학교 생명과학부)
  • Published : 2002.04.01

Abstract

Bacillus polyfermenticus SCD, which is commonly called a 'Bisroot' strain, has been appropriately used for the treatment of long-term intestinal disorders, since the live strains, in the form of active endospores, can successfully reach the target intestine. Goal of this study was to develop an industrial medium for growth and sporulation of B. polyfermenticus SCD. From the results of effect of mixed carbon sources on growth and sporulation of B. polyfermenticus SCD, glucose 2% and starch 2% was particularly found to be the most effective for the maximum number of spore production, resulting in spore cells of $4.3{\times}10^9\;spores/mL$ with a sporulation yield of 91%. For the effect of nitrogen sources, the maximum spore cells of $5.7{\times}10^9\;spores/mL$ of B. polyfermenticus SCD with a sporulation yield of 97% was obtained when B. polyfermenticus SCD was cultivated in an optimum nitrogen source medium containing 5% soybean flour. A medium involving proper phosphate salt yielded the maximum number of a spore cells of $6.0{\times}10^9\;spores/mL$ with a sporulation yield of 95%. Finally, the efficacy of an industrial medium (KH5 medium) on growth and sporulation of B. polyfermenticus SCD was investigated in jar fermenter. The higher number of viable cells $(3.3{\times}10^{10}\;cells/mL)$ and spore cells $(3.0{\times}10^{10}\;spores/mL)$ were obtained in 5 L fermenter when compared with a 500 mL baffle flask cultivation. Thus, KH5 medium developed in this study shows promise as an industrial medium because of higher cells and sporulation yield.

본 연구에서는 프로바이오틱 생균인 비스루트균의 최적 배지 탐색을 위해 탄소원, 질소원 그리고 아포형성의 극대화를 위한 인산염의 최적 농도를 검토하였다. 500 mL baffle flask 에서 배양을 실시한 결과, 탄소원 영향 실험에서는 포도당 2%(w/v)와 전분 2%(w/v)가 첨가되었을 때 최대 총균수$(4.7{\times}10^9\;CFU/mL)$와 활성아포수$(4.3{\times}10^9\;CFU/mL)$를 보였으며 아포형성율은 91%이었다. 질소원 영향 실험에서는 대두분이 5%(w/v) 첨가되었을 때 최대 총균수$(5.9{\times}10^9\;CFU/mL)$와 활성아포수$(5.7{\times}10^9\;CFU/mL)$를 나타내었다. 그리고, 인산염 실험에서는 최대 총균수$(6.3{\times}10^9\;CFU/mL)$와 활성아포수$(6.0{\times}10^9\;CFU/mL)$$KH_2PO_4$를 1%(w/v) 첨가되었을 때 얻을 수 있었으며 아포형성율은 95%를 보였다. 5L 발효조에서 최적 배지인 KH5 배지를 이용하여 $32^{\circ}C$의 온도에서 통기량 1 vvm, 교반속도 450 rpm, pH $7.0{\pm}0.1$로 맞추어 회분식 배양을 실시한 결과 최대 총균수는 $3.3{\times}10^{10}\;CFU/mL$을 보였으며 최대 활성아포수는 $3.0{\times}10^{10}\;CFU/mL$을 나타내었고, 아포형성율은 94%를 나타내었다. 이때 아포 생산성은 $5{\times}10^8\;spores/mL/h$이었다.

Keywords

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