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Expression of cytochrome P‐450 d by Saccharomyces cerevisiae
Author(s) -
Shimizu Toru,
Sogawa Kazuhiro,
Fujii-Kuriyama Yoshiaki,
Takahashi Masae,
Ogoma Yoshiro,
Hatano Masahiro
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)81491-0
Subject(s) - shuttle vector , cytochrome , yeast , saccharomyces cerevisiae , microbiology and biotechnology , microsome , complementary dna , biochemistry , escherichia coli , biology , chemistry , enzyme , gene , recombinant dna , vector (molecular biology)
Rat liver microsomal cytochrome P‐450 d was abundantly expressed in the yeast Saccharomyces cerevisiae by using a yeast‐ Escherichia coli shuttle vector consisting of rat liver P‐450 d cDNA and yeast acid phosphatase promoter. The expressed cytochrome P‐450 d was immunologically crossed with rat liver P‐450 d . The hydroxylase activity of estra‐1,3,5(10)‐triene‐3,17β‐diol was 11 nmol/min per nmol P‐450 d , which is comparable to that reported previously for rat liver P‐450 d . The expressed P‐450 d content was nearly 1% of total yeast protein as estimated from immunoblotting, hydroxylase activity and optical absorption of the reduced CO form.