Purification and characterization of three parathion hydrolases from gram-negative bacterial strains.
Three unique parathion hydrolases were purified from gram-negative bacterial isolates and characterized. All three purified enzymes had roughly comparable affinities for ethyl parathion and had broad temperature optima at ca. 40 degrees C. The membrane-bound hydrolase of Flavobacterium sp. strain ATCC 27551 was composed of a single subunit of approximately 35,000 daltons (Da) and was inhibited by sulfhydryl reagents such as dithiothreitol (DTT) and by metal salts such as CuCl2. The cytosolic hydrolase of strain B-1 was composed of a single subunit of approximately 43,000 Da and was stimulated by DTT and inhibited by CuCl2. The membrane-bound hydrolase of strain SC was composed of four identical subunits of 67,000 Da and was inhibited by DTT and stimulated by CuCl2. The substrate ranges of the three enzymes also differed, as evidenced by their relative affinities for parathion and the related organophosphate insecticide O-ethyl-O-4-nitrophenyl phenylphosphonothioate (EPN). The B-1 hydrolase displayed equal affinity for both compounds, the Flavobacterium enzyme showed twofold-lower affinity for EPN than for parathion, and the SC hydrolase displayed no activity toward EPN. The range in characteristics of these three enzymes can be exploited in different waste disposal strategies.
Mulbry
W W
WW
Pesticide Degradation Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705.
Karns
J S
JS
D043303:Aryldialkylphosphatase; D002462:Cell Membrane / Q000201:enzymology; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D005417:Flavobacterium / Q000201:enzymology*; D006090:Gram-Negative Bacteria / Q000201:enzymology; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D013045:Species Specificity
D007527:Isoenzymes; D046911:Macromolecular Substances; D010744:Phosphoric Monoester Hydrolases; D043303:Aryldialkylphosphatase