Korean Chemical Engineering Research

The Korean Institute of Chemical Engineers (한국화학공학회)
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Optimization of Medium for Protease Production by Enterobacteriaceae sp. PAMC 25617 by Response Surface Methodology

반응표면분석법을 통한 Enterobacteriaceae sp. PAMC 25617의 protease 생산배지 최적화

  • Kim, Hyun-do (Department of Biotechnology and Bioengineering, Chonnam National University) ;
  • Yun, Chul-Won (Department of Biotechnology and Bioengineering, Chonnam National University) ;
  • Choi, Jong-il (Department of Biotechnology and Bioengineering, Chonnam National University) ;
  • Han, Se Jong (Division of Life Sciences, Korea Polar Research Institute)
  • 김현도 (전남대학교 생물공학과) ;
  • 윤철원 (전남대학교 생물공학과) ;
  • 최종일 (전남대학교 생물공학과) ;
  • 한세종 (극지연구소 극지생명과학연구부)
  • Received : 2014.09.29
  • Accepted : 2014.11.24
  • Published : 2015.08.01
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Abstract

This study was conducted to optimize the medium composition for cold-adaptive protease production of Enterobacteriaceae sp. by response surface methodology (RSM). Yeast extract, and TritonX-100 were identified as the significant factors affecting protease from one-factor-at-a-time method. RSM studies for optimizing protease production of Enterobacteriaceae sp. have been carried out for three parameters including yeast extract concentration, TritonX-100 concentration, and culture pH. These significant factors were optimized as 6.690 g/L yeast extract, 0.018 g/L Triton$^{TM}$ X-10, and pH 6.677. The experimentally obtained protease activity was 8.03 U /L, and it became 1.5-fold increase before optimization.

본 논문에서는 저온활성 protease의 생산을 최적화하기 위하여 극지 미생물인 Enterobacteriaceae sp. PAMC 25617의 반응표면분석법을 이용한 배지의 최적화를 수행하였다. One-factor-at-a-time 방법을 이용하여 yeast extract, TritonX-100이 protease의 생산에 영향을 미치는 주요인자인 것을 확인하였다. 물리적인 환경 요인으로 pH를 추가하여 반응표면분석 방법을 이용한 최대 protease 생산 농도를 갖는 각 인자들의 농도를 확인한 결과 5 g/L peptone, 3 g/L malt extract, 10 g/L $C_6H_{12}O_6$, 6.690 g/L yeast extract, 0.018 g/L TritonX-100의 농도에 pH 6.777의 조건에서 미생물을 배양하였을 경우, 최대 10.049 U/L의 protease가 생산될 수 있는 것으로 예측되었다. 실제 배양 결과 8.03 U/L의 protease가 얻어졌으며, 최적화 이전의 생산농도와 비교하여 150% 이상의 증가를 이루었다. 결과적으로 배지최적화를 통한 protease 생산량의 증가에 반응표면분석법의 적용이 유용하다는 것을 확인할 수 있는다. 이러한 결과로부터, 배지 최적화를 이용한 극지 미생물 유래 cold-adapted protease 생산량의 증가가 여러 산업 분야에서 유용하게 이용될 수 있을 것으로 생각된다.

Keywords

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