Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production of Cyanocarboxylic Acid by Acidovorax facilis 72W Nitrilase Displayed on the Spore Surface of Bacillus subtilis

  • Zhong, Xia (College of Life Science and Technology, Jinan University) ;
  • Yang, Shaomin (Department of Pain Medicine, Shenzhen Municipal Sixth People's Hospital) ;
  • Su, Xinying (College of Life Science and Technology, Jinan University) ;
  • Shen, Xiaoxia (College of Life Science and Technology, Jinan University) ;
  • Zhao, Wen (College of Chemistry, Biology and Materials Engineering, Suzhou University of Science and Technology) ;
  • Chan, Zhi (College of Chemistry, Biology and Materials Engineering, Suzhou University of Science and Technology)
  • Received : 2019.01.16
  • Accepted : 2019.03.19
  • Published : 2019.05.28
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Abstract

Nitrilase is a valuable hydrolase that catalyzes nitriles into carboxylic acid and ammonia. Its applications, however, are severely restricted by the harsh conditions of industrial reaction processes. To solve this problem, a nitrilase from Acidovorax facilis 72W was inserted into an Escherichia coli-Bacillus subtilis shuttle vector for spore surface display. Western blot, enzyme activity measurements and flow cytometric analysis results all indicated a successful spore surface display of the CotB-nit fusion protein. In addition, the optimal catalytic pH value and temperature of the displayed nitrilase were determined to be 7.0 and $50^{\circ}C$, respectively. Moreover, results of reusability tests revealed that 64% of the initial activity of the displayed nitrilase was still retained at the $10^{th}$ cycle. Furthermore, hydrolysis efficiency of upscale production of cyanocarboxylic acid was significantly higher in the displayed nitrilase-treated group than in the free group expressed by E. coli (pET-28a-nit). Generally, the display of A. facilis 72W nitrilase on the spore surface of Bacillus subtilis may be a useful method for immobilization of enzyme and consequent biocatalytic stabilization.

Keywords

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