Drug Metabolism: Isolation, Expression and Purification of Cytochrome P450

Cytochrome P450 is a ?superfamily? of haemoproteins that catalyze the enzymatic oxidation of both exogenous and endogenous compounds. One important function of this class of enzymes is to oxidize drugs, making them easier to purge from the body. In the process of oxidation, secondary metabolites are produced which may or may not be more toxic to the host organism. The focus of our research is to (i) isolate new P450 gene sequences from Actinoplanes sp. 53771 and (ii) successfully express cloned P450 genes in E. coli and P. putida and purify the P450 enzymes for subsequent experimentation. We were able to find the complete gene sequence of one new P450, CYPPA3, and two ferredoxins, FdxA1 and FdxA2, from Actinoplanes sp. 53771; locate parts of the gene sequence of two new P450s, CYPPA4 and CYPPA5; and successfully express the cloned P450, PA1, in E. coli and P. putida.