Purification and properties of a second enzyme catalyzing the splitting of carbon-mercury linkages from mercury-resistant Pseudomonas K-62.
An enzyme (splitting enzyme 2) which catalyzes the splitting of carbon-mercury linkage of arylmercury compounds was found in extracts of mercury-resistant Pseudomonas K-62. This enzyme was purified about 725-fold by treatment with streptomycin, precipitation with ammonium sulfate, and successive chromatography on Sephadex G-75 and diethylaminoethyl-cellulose. A purified preparation of the enzyme showed a single band in electrophoresis either on polyacrylamide or sodium dodecyl sulfate-containing polyacrylamide gels. The molecular weight of the enzyme was estimated to be 20,000 (determined by Sephadex G-75 gel filtration) 17,000 (determined by sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis). The enzyme showed a Km of 180 micron and a Vmax of 3.1 mumol/min per mg for p-chloromercuribenzoic acid and a Km of 250 micron and a Vmax of 20 mumol/min per mg for phenylmercuric acetate. The optimum temperature and pH for the reaction were 40 degrees C and 5.0, respectively.
D002729:Chloromercuribenzoates / Q000378:metabolism; D004352:Drug Resistance, Microbial; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000302:isolation & purification; D008628:Mercury / Q000494:pharmacology; D008970:Molecular Weight; D010088:Oxidoreductases / Q000378:metabolism; D010663:Phenylmercury Compounds / Q000378:metabolism*; D011549:Pseudomonas / Q000187:drug effects / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D002729:Chloromercuribenzoates; D010663:Phenylmercury Compounds; D013438:Sulfhydryl Compounds; D010088:Oxidoreductases; D006867:Hydrolases; D008628:Mercury