ADENOSYLCOBALAMIN-DEPENDENT METHYLMALONYL-COA MUTASE FROM PROPIONIBACTERIUM-SHERMANII - ACTIVE HOLOENZYME PRODUCED FROM ESCHERICHIA-COLI

被引：47

作者：

MCKIE, N ^{[1
]}

KEEP, NH ^{[1
]}

PATCHETT, ML ^{[1
]}

LEADLAY, PF ^{[1
]}

机构：

[1] UNIV CAMBRIDGE,DEPT BIOCHEM,TENNIS COURT RD,CAMBRIDGE CB2 1QW,ENGLAND

来源：

BIOCHEMICAL JOURNAL | 1990年 / 269卷 / 02期

基金：

英国惠康基金;

关键词：

D O I：

10.1042/bj2690293

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：

The linked structural genes coding for both subunits of adenosylcobalamin-dependent methylmalonyl-CoA mutase from the Gram-positive bacterium Propionibacterium shermanii have been altered by site-directed mutagenesis and placed under the control of an inducible phage-T7-specific plasmid promoter in Escherichia coli. Conditions have been found under which both α- and β-subunits are produced in soluble form, in near 1:1 ratio, and assemble to form apo-mutase totalling about 5% of the total cellular protein. Methylmalonyl-CoA mutase purified from these cells could be readily converted into the holoenzyme by addition of adenosylcobalamin. The active holoenzyme apparently crystallizes in the same space group as an inactive corrinoid-containing form of the enzyme obtained previously.

引用

页码：293 / 298

页数：6