Highly purified biotin synthase can transform dethiobiotin into biotin in the absence of any other protein, in the presence of photoreduced deazaflavin

被引:49
作者
Mejean, A
Bui, BTS
Florentin, D
Ploux, O
Izumi, Y
Marquet, A
机构
[1] UNIV PARIS 06,LAB CHIM ORGAN BIOL,URA CNRS 493,F-75252 PARIS 05,FRANCE
[2] TOTTORI UNIV,FAC ENGN,DEPT BIOTECHNOL,TOTTORI 680,JAPAN
关键词
D O I
10.1006/bbrc.1995.2900
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
Biotin synthase from Bacillus Sphaericus has been purified to homogeneity from a recombinant strain. The UV-visible spectrum of the pure protein reveals the presence of a [2Fe-2S] cluster. The enzyme is active in the conversion of dethiobiotin to biotin in vitro, in the presence of NADPH, AdoMet and additional unidentified components from the I crude extract of B. sphaericus wild type. We have also found that photoreduced deazaflavin can substitute for the crude extract and NADPH. In this system, biotin synthase is capable of transforming dethiobiotin into biotin in the absence of any other protein but at a substoichiometric level. When this assay was conducted in the presence of [S-35]cysteine, no S-35 was incorporated into biotin, contrary to what happens in the presence of the crude extract. (C) 1995 Academic Press. Inc.
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页码:1231 / 1237
页数:7
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