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A Spectrophotometric Assay for Cytochrome P450 Monooxygenase Activity

Lee, Sung-Eun   (Department of Environmental Science, Policy and Management, University of CaliforniaCC0069995  ); Choi, Won-Sik   (Department of Life Science, Soonchunhyang UniversityUU0000848  ); Park, Byeoung-Soo   (Department of Life Science, Soonchunhyang UniversityUU0000848  ); Lee, Byung-Ho   (Department of Life Science, Soonchunhyang UniversityUU0000848  );
  • 초록

    An assay for cytochrome P450 monooxygenase activity by determination of the products of organophosphate oxidation via inhibition of acetylcholinesterase was described. Extracts from strains of Oryzaephilus surinamensis selected for resistance to chlorpyrifos-methyl (QVOS 102), fenitrothion (VOS F) and malathion (VOS 3), and a standard susceptible strain VOS 48, were incubated with an organophosphate in the presence of a NADPH-generating system and acetylcholinesterase. The degree of inhibition of the acetylchoinesterase activity was converted to manooxygenase activity using standard curves for the inhibition of acetylcholiesterase by chlorpyrifos-methyl-oxon, fenitrooxon and malaoxan. Activity was detectable in VOS 48 and was present at different increased levels with the different organophosphates in the three resistant strains, suggesting that different forms of P450 might be involved in organophosphate oxidation in these insects. The assays were carried out using crude insect homogenates and much smaller samples of insect material than the standard aldrin to dieldrin assay. It should be possible to use the method for determination of monooxygenase activity in single insert.


  • 주제어

    Cytochrome P450 .   monooxygenase activity .   aldrin epoxidation activity .   organophosphate insecticides.  

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  • Choi, Won-Sik (36)

  • Park, Byeoung-Soo (3)

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