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Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

Science in China Series C: Life Sciences volume 51, pages 52–59 (2008)Cite this article

Abstract

The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg−1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65°C and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65 °C.

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Affiliations

  1. College of Engineering, Shenyang Agricultural University, Shenyang, 110161, China

    Zhang Min

  2. College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China

    Zhang Min, Zhao Cong, Du LianXiang & Lu FuPing

  3. College of Life Science, Nankai University, Tianjin, 300071, China

    Gao Chen

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  1. Zhang Min
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  2. Zhao Cong
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  3. Du LianXiang
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  4. Lu FuPing
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  5. Gao Chen
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Correspondence to Lu FuPing.

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Supported by the “863” High-Tech Research Project of China (Grant No. 2007AA 02Z212)

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Zhang, M., Zhao, C., Du, L. et al. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis . Sci. China Ser. C-Life Sci. 51, 52–59 (2008). https://doi.org/10.1007/s11427-008-0009-9

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  • DOI: https://doi.org/10.1007/s11427-008-0009-9

Keywords

  • Bacillus subtilis
  • thermophilic neutral protease
  • expression
  • purification
  • characterization