Purification and characterization of carbaryl hydrolase from Blastobacter sp. strain M501

A bacterium capable of hydrolyzing carbaryl (1-naphthyl-N-methylcarbamate) was isolated from a soil enrichment. This bacterium was characterized taxonomically as a Blastobacter sp. and designated strain M501. A carbaryl hydrolase present in this strain was purified to homogeneity by protamine sulfate treatment, ammonium sulfate precipitation, and hydrophobic, anion-exchange, gel filtration, and hydroxylapatite chromatographies. The native enzyme had a molecular mass of 166,000 Da and was composed of two subunits with molecular masses of 84,000 Da. The optimum pH and temperature of the enzyme activity were 9.0 and 45 degrees C, respectively. The enzyme was not stable at temperatures above 40 degree C. The purified enzyme hydrolyzed seven N-methylcarbamate insecticides and also exhibited activity against 1-naphthyl acetate and 4-nitrophenyl acetate