Highly Purified Biotin Synthase Can Transform Dethiobiotin into Biotin in the Absence of Any Other Protein, in the Presence of Photoreduced Deazaflavin

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...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1995-12, Vol.217 (3), p.1231-1237
Main Authors: Mejean, A., Bui, B.T.S., Florentin, D., Ploux, O., Izumi, Y., Marquet, A.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacillus - enzymology
Biotin - analogs & derivatives
Biotin - metabolism
Cell-Free System
Cysteine - metabolism
Flavins - metabolism
Molecular Sequence Data
Spectrum Analysis
Sulfur - metabolism
Sulfurtransferases - isolation & purification
Sulfurtransferases - metabolism
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Summary: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 [35S]cysteine, no 35S was incorporated into biotin, contrary to what happens in the presence of the crude extract.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1995.2900