CLONING SEQUENCING AND EXPRESSION OF THE GENE FOR CYTOCHROME-P450MEG, THE STEROID-15-BETA-MONOOXYGENASE FROM BACILLUS-MEGATERIUM ATCC-13368

被引：35

作者：

RAUSCHENBACH, R ^{[1
]}

ISERNHAGEN, M ^{[1
]}

NOESKEJUNGBLUT, C ^{[1
]}

BOIDOL, W ^{[1
]}

SIEWERT, G ^{[1
]}

机构：

[1] SCHERING AG,RES LABS,MULLERSTR 178,D-13342 BERLIN,GERMANY

来源：

MOLECULAR & GENERAL GENETICS | 1993年 / 241卷 / 1-2期

关键词：

CYTOCHROME-P450; STEROID HYDROXYLATION; BACILLUS-MEGATERIUM; BACILLUS-SUBTILIS;

D O I：

10.1007/BF00280214

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A 4.3 kb EcoRI fragment carrying the gene for cytochrome P450meg, the steroid-15beta-monooxygenase from Bacillus megaterium ATCC 13368, was cloned and completely sequenced. The gene codes for a protein of 410 amino acids and was expressed in Escherichia coli and B. subtilis. Protein extracts from the recombinant E. coli strains were able to hydroxylate corticosteroids in the 15beta position when supplemented with an extract from a P450- mutant of B. megaterium ATCC 13368 as a source of megaredoxin and megaredoxin reductase. In contrast, 15beta-hydroxylation was obtained in vitro and in vivo without the addition of external electron transfer proteins, when cytochrome P450meg was produced in B. subtilis 168. Protein extracts from nonrecombinant B. subtilis 168 could also support the in vitro hydroxylation by cytochrome P450meg produced in E. coli.

引用

页码：170 / 176

页数：7