Journal of Applied Biological Chemistry

  • 제66권
  • /
  • Pages.128-137
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  • 2023
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  • 1976-0442(pISSN)
  • /
  • 2234-7941(eISSN)
한국응용생명화학회 (The Korean Society for Applied Biological Chemistry)
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Novel sinIR promoter for Bacillus subtilis DB104 recombinant protein expression system

  • Ji-Su Jun (Department of Food Science and Biotechnology, College of Life Science and Biotechnology, Dongguk University) ;
  • Min-Joo Kim (Department of Food Science and Biotechnology, College of Life Science and Biotechnology, Dongguk University) ;
  • KwangWon Hong (Department of Food Science and Biotechnology, College of Life Science and Biotechnology, Dongguk University)
  • 투고 : 2023.02.27
  • 심사 : 2023.03.20
  • 발행 : 2023.12.31
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초록

Transcriptome analysis revealed that the sinR gene encoding a transition-state regulator of Bacillus pumilus, genetically close to B. subtilis, was expressed at high levels during growth. The sinR gene is the second gene of the sinIR operon consisting of three promoters and two structural genes in B. subtilis. This study used the sinIR promoter of B. subtilis DB104 to construct a recombinant protein expression system. First, the expression ability depending on the number of sinIR promoter was investigated using enhanced green fluorescent protein (eGFP). The expression level of eGFP was slightly higher when using two promoters (Psin2) than using original promoters. The Psin2 promoter was further engineered by modifying the repressor binding site and -35 and -10 regions. Shine-Dalgarno (SD) sequence of the sinI gene was modified to the consensus sequence. Finally, combining the engineered Psin2 promoter with the modified SD sequence increased the expression level of eGFP by about 13.4-fold over the original promoter. Our results suggest that the optimized sinIR promoter could be used as a novel tool for recombinant protein expression in B. subtilis.

키워드

과제정보

This research was supported by the Basic Science Research Programme through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2021R1I1A2046284).

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