Abstract
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.
MeSH terms
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Aryl Hydrocarbon Hydroxylases / biosynthesis*
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Aryl Hydrocarbon Hydroxylases / chemistry
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Aryl Hydrocarbon Hydroxylases / genetics
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Aryl Hydrocarbon Hydroxylases / isolation & purification
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Bacterial Proteins / biosynthesis*
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Bacterial Proteins / genetics
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Chaperonin 10 / biosynthesis*
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Chaperonin 10 / genetics
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Chaperonins
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Coumarins / chemistry
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Cytochrome P-450 CYP2B6
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Escherichia coli Proteins
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Escherichia coli*
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Gene Expression*
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Heat-Shock Proteins / biosynthesis*
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Heat-Shock Proteins / genetics
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Humans
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Oxidoreductases, N-Demethylating / biosynthesis*
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Oxidoreductases, N-Demethylating / chemistry
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Oxidoreductases, N-Demethylating / genetics
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Oxidoreductases, N-Demethylating / isolation & purification
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Recombinant Proteins / biosynthesis*
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
Substances
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Bacterial Proteins
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Chaperonin 10
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Coumarins
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Escherichia coli Proteins
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GroE protein, E coli
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Heat-Shock Proteins
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Recombinant Proteins
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7-ethoxy-4-trifluoromethylcoumarin
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Aryl Hydrocarbon Hydroxylases
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CYP2B6 protein, human
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Cytochrome P-450 CYP2B6
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Oxidoreductases, N-Demethylating
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Chaperonins