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Continuous Regeneration of NAD(H) Covalently Bound to a Cysteine Genetically Engineered into Glucose Dehydrogenase

Bio/Technology volume 9, pages 280–284 (1991)Cite this article

Abstract

We introduced a cysteine residue on the surface of glucose dehydrogenase from Bacillus subtilis using site-directed muta-genesis. To this mutant, an NAD-analogue was covalently attached by a disulphide bridge so that it was active intramolecularly. The glucose dehydrogenase-cys44-NAD complex, which contained one reactive NAD molecule per subunit of glucose dehydrogenase, was operated together with lactate dehydrogenase in a coupled enzymatic regeneration of NAD(H) in a hollow fiber reactor. L-lactate and gluconic acid were continuously produced from pyruvate and D-glucose, respectively, with a turnover number of 45 cycles per minute for each NAD molecule. The total turnover per coenzyme was 135,000 for the first 2.5 days.

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Author notes

  1. Klaus Mosbach: Corresponding author.

Authors and Affiliations

  1. Pure and Applied Biochemistry, Chemical Center, University of Lund, P.O. Box 124, S-221 00, Lund, Sweden

    Mats Persson, Mats-Olle Månsson, Leif Bülow & Klaus Mosbach

Authors
  1. Mats Persson
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  2. Mats-Olle Månsson
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  3. Leif Bülow
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  4. Klaus Mosbach
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Persson, M., Månsson, MO., Bülow, L. et al. Continuous Regeneration of NAD(H) Covalently Bound to a Cysteine Genetically Engineered into Glucose Dehydrogenase. Nat Biotechnol 9, 280–284 (1991). https://doi.org/10.1038/nbt0391-280

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  • DOI: https://doi.org/10.1038/nbt0391-280

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