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European Journal of Biochemistry pp 102–109Cite as

Reduced Nicotinamide Dinucleotide Phosphate Diaphorase from Bacillus subtilis

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Abstract

NADPH-diaphorase was extensively purified from extracts of Bacillus subtilis NY. FMN is most probably the prosthetic group of this enzyme. NADPH is oxidized by oxygen as the terminal acceptor only in the presence of catalytic amounts of free flavins (K m for FMN was 1.6 × 10−6 M at pH 8.0). NADPH was directly oxidized in the presence of various electron acceptors such as quinones, ferricyanide and tetrazolium salts. Cytochrome c was a very poor acceptor in this system. K m for NADPH was 2.0 × 10−4M and K i for NADP+ as a competitive inhibitor was 1.1 × 10−3 M at pH 7.2. Mn++ or Mg++ at mM concentration showed some stimulation of enzyme activity. The enzyme was partially inhibited by various chelating agents and p-hydroxymercuribenzoate but not by several typical inhibitors of the respiratory chain. It is suggested that this flavoprotein can serve as a useful tool for NADP+ regeneration in coupled enzyme reactions.

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Abbreviations

DCPIP:

2,6-dichlorophenolindophenol

INT:

2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride

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Authors and Affiliations

  1. Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israël

    G. Avigad & N. Levin

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  1. G. Avigad
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  2. N. Levin
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Editors and Affiliations

  1. Liège, Belgium

    Claude Liébecq

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Avigad, G., Levin, N. (1967). Reduced Nicotinamide Dinucleotide Phosphate Diaphorase from Bacillus subtilis . In: Liébecq, C. (eds) European Journal of Biochemistry. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-25813-2_17

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  • DOI: https://doi.org/10.1007/978-3-662-25813-2_17

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-23717-5

  • Online ISBN: 978-3-662-25813-2

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