Overexpression of the coenzyme-F-420-dependent N-5,N-10-methylenetetrahydromethanopterin dehydrogenase gene from the hyperthermophilic Methanopyrus kandleri

被引:19
作者
Klein, AR
Thauer, RK
机构
[1] MAX PLANCK INST TERR MIKROBIOL,D-35043 MARBURG,GERMANY
[2] UNIV MARBURG,FACHBEREICH BIOL,MIKROBIOL LAB,D-3550 MARBURG,GERMANY
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1997年 / 245卷 / 02期
关键词
hyperthermophilic enzyme; thermostability; lyotropic salts; heterologous expression; methanogenic Archaea;
D O I
10.1111/j.1432-1033.1997.t01-1-00386.x
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 mtd gene encoding coenzyme-F-420-dependent N-5,N-10-methylenetetrahydromethanopterin dehydrogenase (Mtd) in the hyperthermophilic Methanopyrus kandleri has been cloned, sequenced and functionally overexpressed in Escherichia cell. The overproduced enzyme was purified in a 90% yield to apparent homogeneity by means of only one chromatographic step. Its thermostability properties and most of its catalytic properties were the. same as those of the native enzyme purified directly from M. kandleri. Only the dependence of the activity on the concentration of lyotropic salts differed slightly. Northern blot analysis revealed that in M kandleri the mtd gene is monocistronically transcribed.
引用
收藏
页码:386 / 391
页数:6
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