ENHANCED INVIVO MONOOXYGENASE ACTIVITIES OF MAMMALIAN P450S IN ENGINEERED YEAST-CELLS PRODUCING HIGH-LEVELS OF NADPH-P450 REDUCTASE AND HUMAN CYTOCHROME-B5

被引:154
作者
TRUAN, G
CULLIN, C
REISDORF, P
URBAN, P
POMPON, D
机构
[1] CNRS,CTR GENET MOLEC,F-91198 GIF SUR YVETTE,FRANCE
[2] UNIV PARIS 11,INST GENET & MICROBIOL,F-91406 ORSAY,FRANCE
关键词
BIOCONVERSION; DRUG METABOLISM; HETEROLOGOUS EXPRESSION; SACCHAROMYCES-CEREVISIAE;
D O I
10.1016/0378-1119(93)90744-N
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
We have engineered yeast genomic DNA to construct a set of strains producing various relative amounts of yeast NADPH-P450 reductase (Yred) and human cytochrome b5 (Hb5). Expression of cDNAs encoding human P450 1A1, 1A2, 3A4, 19A and mouse P450 1A1 in the different oxido-reduction backgrounds thus constituted were achieved after strain transformation by plasmid-based P450-encoding expression cassettes. The results indicate that the level of Yred strongly affects all activities tested. In contrast, the amount of Hb5 affects activities in a manner that is dependent both on the P450 isoform considered and the Yred level. In a strain containing optimized amounts of Hb5 and Yred, human P450 3A4-specific testosterone-6beta-hydroxylase activity can be enhanced as much as 73-fold in comparison with the activity observed in a wild-type strain. Bioconversion of sterols or xenobiotics was easily achieved in vivo using this new co-expression system.
引用
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页码:49 / 55
页数:7
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