Improvement in the expression of CYP2B6 by co-expression with molecular chaperones GroES/EL in Escherichia coli.
Improvement of CYP2B6 expression was examined by co-expression with molecular chaperones GroES/EL. Although a CO-reduced difference spectrum was not detected in Escherichia coli transformed only by the CYP2B6-expressing vector, co-expression of GroES/EL resulted in high-level expression which reached over 2000 nmol P450/L. CYP2B6 was purified from the E. coli membrane with a high yield. Purified CYP2B6 showed 7-ethoxy-4-trifluoromethylcoumarin O-deethylase activity in a reconstitution system. This expression system would be useful for the production of large amounts of active CYP2B6 and for the detailed analysis of the enzyme.
Mitsuda
Maori
M
Department of Biology, Graduate School of Science, Osaka University, 2-17-85, Jusohonmachi, Osaka 532-8686, Japan.
Iwasaki
Masahiko
M
D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D018835:Chaperonin 10 / Q000096:biosynthesis* / Q000235:genetics; D018833:Chaperonins; D003374:Coumarins / Q000737:chemistry; D065702:Cytochrome P-450 CYP2B6; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D018835:Chaperonin 10; D003374:Coumarins; D029968:Escherichia coli Proteins; C028715:GroE protein, E coli; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; C056033:7-ethoxy-4-trifluoromethylcoumarin; D001189:Aryl Hydrocarbon Hydroxylases; C585599:CYP2B6 protein, human; D065702:Cytochrome P-450 CYP2B6; D010089:Oxidoreductases, N-Demethylating; D018833:Chaperonins