• DOI:10.1007/S12033-007-0060-Y
• Corpus ID: 11595829

Secretory Expression of Nattokinase from Bacillus subtilis YF38 in Escherichia coli

@article{Liang2007SecretoryEO,
  title={Secretory Expression of Nattokinase from Bacillus subtilis YF38 in Escherichia coli},
  author={Xiaobo Liang and Shifang Jia and Yufang Sun and Meiling Chen and Xiuzhu Chen and Jin Zhong and Liandong Huan},
  journal={Molecular Biotechnology},
  year={2007},
  volume={37},
  pages={187-194}
}

• Xiaobo Liang, S. Jia, +4 authors Liandong Huan
• Published 17 July 2007
• Biology
• Molecular Biotechnology

Nattokinase producing bacterium, B. subtilis YF38, was isolated from douchi, using the fibrin plate method. The gene encoding this enzyme was cloned by polymerase chain reaction (PCR). Cytoplasmic expression of this enzyme in E. coli resulted in inactive inclusion bodies. But with the help of two different signal peptides, the native signal peptide of nattokinase and the signal peptide of PelB, active nattokinase was successfully expressed in E. coli with periplasmic secretion, and the nattokinase in culture medium displayed high fibrinolytic activity. The fibrinolytic activity of the expressed enzyme in the culture was determined to reach 260 urokinase units per micro-liter when the recombinant strain was induced by 0.7 mmol l−1 isopropyl-β-D- thiogalactopyranoside (IPTG) at 20°C for 20 h, resulting 49.3 mg active enzyme per liter culture. The characteristic of this recombinant nattokinase is comparable to the native nattokinase from B. subtilis YF38. Secretory expression of nattokinase in E. coli would facilitate the development of this enzyme into a therapeutic product for the control and prevention of thrombosis diseases.