Production of Cyanocarboxylic Acid by Acidovorax facilis 72W Nitrilase Displayed on the Spore Surface of Bacillus subtilis.
J Microbiol Biotechnol
; 29(5): 749-757, 2019 May 28.
Article
em En
| MEDLINE
| ID: mdl-30955259
ABSTRACT
Nitrilase is a valuable type of 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°C, respectively. Moreover, results of reusability tests revealed that 64% of the initial activity of the displayed nitrilase was still retained at the 10th 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.
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01-internacional
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MEDLINE
Assunto principal:
Esporos Bacterianos
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Bacillus subtilis
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Comamonadaceae
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Aminoidrolases
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article