Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa (Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Department of Microbiology, Kyungpook National University) ;
  • Song, Jae-Jun (Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Choi, Yoon-Ho (BioLeaders Corp.) ;
  • Hong, Seung-Pyo (BioLeaders Corp.) ;
  • Rha, Eu-Gene (Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Hyung-Kwoun (Division of Biosciences and Technology, Catholic University) ;
  • Lee, Seung-Goo (Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Poo, Har-Young (Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Sang-Chul (Laboratory of Microbial Function, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Seu, Young-Bae (Department of Microbiology, Kyungpook National University) ;
  • Sung, Moon-Hee (BioLeaders Corp., Department of Bio & Nonochemistry, Kookmin University)
  • Published : 2003.12.01
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Abstract

High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

Keywords

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