Purification and characterization of carbaryl hydrolase from Arthrobacter sp. RC100.
A carbaryl hydrolase was purified to homogeneity from Arthrobacter sp. strain RC100 by protamine sulfate treatment, ammonium sulfate precipitation, and hydrophobic, anion-exchange, and gel filtration chromatographies. The native enzyme had a molecular mass of 100 kDa and was composed of two identical subunits with molecular masses of 51 kDa. The hydrolase activity was strongly inhibited by DIFP, PMSF, Hg(2+) and paraoxon but not by EDTA. The optimum pH and temperature for the enzyme activity were 9.0 and 50 degrees C, respectively. The enzyme hydrolyzed four N-methylcarbamate insecticides (carbaryl, xylylcarb, metolcarb and XMC), but was not able to hydrolyze fenobucarb, propoxur, and isoprocarb.
Hayatsu
M
M
Faculty of Agriculture, Shizuoka University, Japan. ahmhaya@agr.shizuoka.ac.jp
Mizutani
A
A
Hashimoto
M
M
Sato
K
K
Hayano
K
K
D001173:Arthrobacter / Q000201:enzymology*; D012721:Carbaryl / Q000378:metabolism*; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007306:Insecticides / Q000378:metabolism*; D007700:Kinetics; D008628:Mercury / Q000494:pharmacology; D008970:Molecular Weight; D010261:Paraoxon / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D021122:Protein Subunits; D013379:Substrate Specificity; D013696:Temperature
D004791:Enzyme Inhibitors; D007306:Insecticides; D011480:Protease Inhibitors; D021122:Protein Subunits; D004492:Edetic Acid; D002265:Carboxylic Ester Hydrolases; C077242:carbamate hydrolase; D008628:Mercury; D010261:Paraoxon; D012721:Carbaryl