Functional expression of human cytochrome P450 enzymes in Escherichia coli.
Knowledge regarding cytochrome P450 (P450) is crucial to the fields of drug therapy and drug development, as well as in our understanding of the mechanisms underlying the metabolic activation of potentially toxic and carcinogenic compounds. Escherichia coli is the most extensively utilized host in the production of recombinant human P450 enzymes. However, the recovery of substantial yields of functionally active P450 proteins remains problematic. Mammalian P450 protein was first expressed in 1991, via the modification of the N-terminal amino acid sequences in E. coli cells. Since that time, a variety of strategies have been established for the functional expression of recombinant P450s in E. coli, including N-terminal modification, the use of molecular chaperones, and culturing at lower temperatures. In all cases, human P450 expressed in E. coli cells has been shown to efficiently catalyze the oxidation of representative substrates at efficient rates. These recombinant P450s are applicable to studies which estimate the kinetic parameters of drug oxidation, and have also been used to determine the metabolic pathways of drugs and carcinogens exploited by human P450s. Despite the potential of P450s in various pharmaceutical and biotechnological fields, however, a host of substantial challenges must be overcome before these enzymes can be routinely utilized. Intrinsically, these enzymes are not very active, and exhibit poor stability. In this review, we have described current developments in the heterologous expression of human P450 enzymes.
Yun
Chul-Ho
CH
School of Biological Sciences and Technology & Hormone Research, Chonnam National University, Gwangju 500-757, Republic of Korea. chyun@jnu.ac.kr
Yim
Sung-Kun
SK
Kim
Dong-Hyun
DH
Ahn
Taeho
T
D018360:Crystallography, X-Ray; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System