Abstract
Mutant-protein production of two variants of Cytochrome P450 were studied. Preliminary rotation-rate electrochemical studies of both wild-type and mutant [1-12G] Cytochrome P450 BM3 from Bacillus magaterium, conducted revealed the variations of the catalytic properties exhibited by this protein. It was concluded that the WT Cytochrome P450 BM3 conducts primarily two-electron transfer of dioxygen [O2] to peroxide, whereas 1-12G catalyses four-electron transfer of dioxygen to water. Thus in orders to further study such variations in catalysis, mutations in other forms of Cytochrome P450 were prepared. Thus, systematic mutagenesis studies of Cytochrome P450 CAM replacing tyrosine 29 and 96 with phenylalanines as well as Cytochrome P450 BM3, in which aspartic acid194 was converted to a cysteine and cysteine397 to asparagine were systematically introduced. Further growth of the before-mentioned variants and future catalytic-rate studies are to be conducted in order to elucidate the catalytic properties of the family Cytochrome P450 in order to ultimately produce a procedure for using such proteins as vehicles for alkane hydroxylation?a key reaction used in various scientific and industrial arenas.
