Abstract
An enzyme which catalyzed the hydrolysis of crotoxyphos ((E)-1-phenylethyl 3-[(dimethoxyphosphinyl)oxy]-2-butenoate) was isolated from nonsterile and radiation-sterilized Chehalis clay loam with 1.5M Tris (2-(hydroxymethyl)-2-nitro-1,3-propanediol) and partially purified with lead acetate treatment. Two soil-g equivalents of lead acetate purified enzyme in pH 8 buffer hydrolyzed 0.13 μmol of substrate to dimethyl phosphate and α-methylbenzyl 3-hydroxycrotonate in 16 hr at 37°C. The enzyme exhibited maximal activity around pH 8.0 and was irreversibly inactivated below pH 5.0 or above pH 10.0. The Km value for crotoxyphos was calculated to be 4.63×10−3 M. The enzyme was stable at 60°C for 10 min, retained activity indefinitely at −10°C, and was completely inactivated within a week at room temperature. When applied to autoclaved Chehalis clay loam, purified enzyme lost 75% of its activity after one week and the remainder within two weeks.
Similar content being viewed by others
References
Chamberlain, W. F.: The metabolism of P32-labeled Shell SD-4294 in a lactating ewe. J. Econ. Entomol.57, 119 (1964a).
—:The metabolism of P32-labeled Ciodrin in a lactating goat. J Econ. Entomol.57, 329 (1964b).
Getzin, L. W., and I. Rosefield: Organophosphorus insecticide degradation by heat-labile substances in soil. J. Agric. Food Chem.16, 598 (1968).
—:Partial purification and properties of a soil enzyme that degrades the insecticide malathion. Biochim. Biophys. Acta235, 442 (1971).
Gibson, W. P., and R. G. Burns: The breakdown of malathion in soil and soil components. Microbial Ecol.3, 219 (1977).
Hanes, C. S., and F. A. Isherwood: Separation of the phosphoric esters on the filter paper chromatogram. Nature164, 1107 (1949).
Iwata, Y., M. Ittig, and F. A. Gunther: Degradation of O,O-dimethyl S-[α-(carboethoxy benzyl] phosphorodithioate (phenthoate) in soil. Arch. Environ. Contam. Toxicol.6, 1 (1977).
Konrad, J. G., and G. Chesters: Degradation in soils of Ciodrin, an organophosphorus insecticide. J. Agric. Food Chem.17, 226 (1969).
Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall: Protein measurement with the Folin phenol reagent. J. Biol. Chem.193, 265 (1951).
Mingelgrin, U., S. Saltzman, and B. Yaron: A possible model for the surface-induced hydrolysis of organophosphorus pesticides on kaolinite clays. Soil Sci. Soc. Am. J.41, 519 (1977).
Mitchell, L. C.: Separation and identification of eleven organo-phosphate pesticides by paper chromatography: Delnav, diazinon, EPN, guthion, malathion, methyl parathion, parathion, phosdrin, ronnel, systox and trithion. J. Assoc. Offic. Anal. Chem.43, 810 (1960).
Satyanarayana, T., and L. W. Getzin: Properties of a stable cell-free esterase from soil. Biochem.12, 1566 (1973).
Skujins, J.: Extracellular enzymes in soil. In A. I. Laskin and H. Lechevalier (ed.): CRC Critical Reviews in Microbiology. Vol. 4, p. 383, Cleveland, OH: CRC Press (1976).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Getzin, L.W., Satyanarayana, T. Isolation of an enzyme from soil that degrades the organophosphorus insecticide, crotoxyphos. Arch. Environ. Contam. Toxicol. 8, 661–672 (1979). https://doi.org/10.1007/BF01054868
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01054868