Zuyu Guo

Purification of functional recombinant P450s from bacteria.

Publisher Summary This chapter discusses the methods for the purification of functional recombinant P450s from bacteria. A number of expression systems have been utilized to produce recombinant P450s, as discussed in this and an earlier volume. I Bacteria have considerable advantages because of the high levels of expression, ease of manipulation, and relatively low cost. While P450s can be studied within some of the other vector systems, purification is usually necessary for most studies with the enzymes produced in bacteria. In general, purification is relatively easy and considerably more efficient than from mammalian tissues Thes chapter focuses on recombinant human P450s expressed in Escherichia coli and, Salmonella typhimurium . The most direct means of monitoring P450 production in bacteria is to do direct difference spectroscopy.

Identification of the Highly Polymorphic S-Mephenytoin Hydroxylase in Humans

The metabolism of the anticonvulsant drug mephenytoin is highly polymorphic in Oriental populations (about 20% of slow metabolizers) in comparison with Caucasians, where 2 – 5% of slow metabolizers are found. It was reported that 4-hydroxylation of the S-enantiomer of mephenytoin is catalyzed by a cytochrome P450 enzyme from the 2C-family. A large number of efforts have been made to characterize this enzyme, but so far its real identity remained unknown.