Overexpression of Xenobiotic Metabolizing Enzymes
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AbstractThe plan is to develop and optimize a method for high level production of functional, catalyticallycytochromes P450 using baculovirus expression in insect cells. Cytochromes P450 are the principal enmetabolism of many drugs, protoxins, and environmental pollutants. This proposed method includes enhcytochrome P450 catalytic activity through the co-expression of cytochrome P450:NADPH oxidoreductasecytochrome b5. Human cytochromes P4502A6, P4502E1, and P4503A4 will be used as prototype enzymes fordevelopment. Catalytic constants, apparent Km, and turnover number will be measured for the optimizeexpressed enzymes and compared to the same enzyme isolated from human liver or produced using otherexpression systems. The eventual goal is to develop a complete panel of baculovirus expressed humanP450 isoforms. The commercial applications of this system would be for the study of human xenobiotic
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