TY - JOUR
T1 - Enhanced catalytic efficiency of nitrilase from Acidovorax facilis 72W and application in bioconversion of 3-cyanopyridine to nicotinic acid
AU - Li, He
AU - Dong, Weiliang
AU - Zhang, Yue
AU - Liu, Kuan
AU - Zhang, Wenming
AU - Zhang, Min
AU - Ma, Jiangfeng
AU - Jiang, Min
N1 - Publisher Copyright:
© 2017
PY - 2016/11/10
Y1 - 2016/11/10
N2 - In this study, the catalytic efficiency using NitA from Acidovorax facilis 72W for nicotinic acid (NA) production was investigated and further improved by site-directed mutagenesis. Results showed that the specific activity of mutated NitA-C2 (F168V-S192F) towards 3-cyanopyridine increased 5-fold to 35 U mg−1 protein. Further characterization of the biochemical properties of both nitrilases showed the optimal pH and temperature were 6.0–8.0 and 60 °C, respectively, whereas the pH and thermal stability of NitA-C2 were decreased. Finally, whole cell catalysis was adopted for NA production and a 100% conversion yield was achieved under 0.1 mol L−1 3-cyanopyridine for both strains. Besides, the conversion rate by E. coli BL21 (DE3-pET-nitA-C2) reached to 1.0 mmol min−1 g−1 wet cell weight, which was 3-fold higher than that by E. coli BL21 (DE3-pET-nitA). These results indicated that the mutated NitA-C2 was a promising candidate which holds potential application in biological NA production.
AB - In this study, the catalytic efficiency using NitA from Acidovorax facilis 72W for nicotinic acid (NA) production was investigated and further improved by site-directed mutagenesis. Results showed that the specific activity of mutated NitA-C2 (F168V-S192F) towards 3-cyanopyridine increased 5-fold to 35 U mg−1 protein. Further characterization of the biochemical properties of both nitrilases showed the optimal pH and temperature were 6.0–8.0 and 60 °C, respectively, whereas the pH and thermal stability of NitA-C2 were decreased. Finally, whole cell catalysis was adopted for NA production and a 100% conversion yield was achieved under 0.1 mol L−1 3-cyanopyridine for both strains. Besides, the conversion rate by E. coli BL21 (DE3-pET-nitA-C2) reached to 1.0 mmol min−1 g−1 wet cell weight, which was 3-fold higher than that by E. coli BL21 (DE3-pET-nitA). These results indicated that the mutated NitA-C2 was a promising candidate which holds potential application in biological NA production.
KW - Acidovorax facilis 72W
KW - Nicotinic acid
KW - Nitrilase
KW - Site-directed mutagenesis
UR - http://www.scopus.com/inward/record.url?scp=85018810697&partnerID=8YFLogxK
U2 - 10.1016/j.molcatb.2017.03.010
DO - 10.1016/j.molcatb.2017.03.010
M3 - 文章
AN - SCOPUS:85018810697
SN - 1381-1177
VL - 133
SP - S459-S467
JO - Journal of Molecular Catalysis - B Enzymatic
JF - Journal of Molecular Catalysis - B Enzymatic
ER -