Study On Drug Intermediates Hydroxylated By Cytochrome P450 BM-3 Multienzyme Combination

Cytochrome P450 BM-3 has broad research prospects in biopharmaceuticals and biosynthesis due to its ability to hydroxylate inert C-H bonds as well as its wide range of catalytic substrates and well stereoselectivity.P450 BM-3 requires NADPH to supply electrons during catalysis while its own electron transport structure is less efficient,limiting its catalytic efficiency.In this paper,the more efficient flavodoxin NADP+oxidoreductase(FNR)was used for electron transport work,and a new catalytic system was constructed with the heme domain of P450 BM-3(P450 BMP).NADPH was regenerated by glucose dehydrogenase(GDH)to form a P450 BMP-FNR-GDH(BFG)multienzyme combined catalytic system.In this paper,the multienzyme catalytic system was immobilized by biomineralization technology to obtain the immobilized enzyme(BFG@Cu)with nanoflower structure.In this paper,the catalytic activity of 1,4-benzodioxane and α-tetralone was studied by using BFG and BFG@Cu.The effects of different biomineralization systems,temperature,and pH on the catalytic reaction were also studied.The results showed that the free BFG had higher catalytic efficiency than before.The immobilized BFG@Cu has higher catalytic efficiency than the BFG,and the stability is also greatly improved at different temperatures and pH.