Purification and characterization of a novel carbaryl hydrolase from Aspergillus niger PY168.
A fungus capable of using carbaryl as the sole source of carbon and energy was isolated from a soil enrichment, and characterized as Aspergillus niger and designated strain PY168. A novel carbaryl hydrolase from cell extract was purified 262-fold to apparent homogeneity with 13.6% overall recovery. It had a monomeric structure with a molecular mass of 50,000 Da and a pI of 4.6, and the enzyme activity was optimal at 45 degrees C and pH 7.5, The activities were strongly inhibited by Hg(2+), Ag+, rho-chloromercuribenzoate, iodoacetic acid, diisofluorophosphate and phenylmethylsulfonyl fluoride but not EDTA and phenanthroline. The purified enzyme hydrolyzed various N-methylcarbamate insecticides. Carbaryl is the preferred substrate.
Zhang
Qing
Q
The Key Laboratory of Gene Engineering of Ministry of Education, Sun Yet-Sen University, Guangzhou 510275, PR China.
Liu
Yang
Y
Liu
Yu-Huan
YH
D001234:Aspergillus niger / Q000201:enzymology*; D012721:Carbaryl / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D007306:Insecticides / Q000378:metabolism*; D019807:Iodoacetic Acid / Q000494:pharmacology; D008670:Metals / Q000494:pharmacology; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013696:Temperature; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
D004791:Enzyme Inhibitors; D007306:Insecticides; D008670:Metals; D013439:Sulfhydryl Reagents; D020245:p-Chloromercuribenzoic Acid; D006867:Hydrolases; D012721:Carbaryl; D019807:Iodoacetic Acid