Purification & characterization of benzonitrile produced by Bacillus pumillus S7

Abstract

Dihalogenate benzonitrile are active compounds in a number of herbicides and poses to have a deleterious health effect. The biocatalyst was applied to the biotransformation of benzonitrile,3-cynopyridine,(R,S)-3-hydroxy -2- ethylene butanenitrile The enzyme involved in the degradation of benzonitrile by bacteria identified as Bacillus pumilus S7a strain which utilized benzonitrile as sole source of carbon and nitrogen were partially purified by ammonium sulphate precipitation of (80%). The GC results revealed that benzonitrile in single step pathway was converted to ammonia with the formation of benzoic acid as an intermediate by enzyme benzonitrilase. In the present study nitrilase enzyme was produced and purified using Bacillus pumilusS7. The purified enzyme was showing maximum activity at a pH 6(35.32 μmole/min) and maximum benzonitrile degradation with free enzyme was observed at temperature 300c (29.27μmole/min).The metal source FeSo4 (40.55 μmole/min) and enzyme stable up to 30 min (45.14 μmole/min) The Vmax is 0.188μg/ml and km is 0.00178μm.The molecular weight of benzonitrilase were found to be 97400 Dalton.