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Characterization and Engineering of a Novel Pyrroloquinoline Quinone Dependent Glucose Dehydrogenase from Sorangium cellulosum So ce56

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Abstract

A novel pyrroloquinoline quinone dependent glucose dehydrogenase like enzyme (PQQ GDH) was isolated from Sorangium cellulosum So ce56. The putative coding region was cloned, over expressed in E. coli and the resulting enzyme was characterized. The recombinant protein has a relative molecular mass of 63 kDa and shows 43% homology to PQQ GDH-B from Acinetobacter calcoaceticus. In the presence of PQQ and CaCl2 the enzyme has dehydrogenase activity with the substrate glucose as well as with other mono- and disaccharides. The thermal stability and its pH activity profile mark the enzyme as a potential glucose biosensor enzyme. In order to decrease the activity on maltose, which is unwanted for a potential application in biosensors, the protein was rationally modified at three specified positions. The best variant showed a 59% reduction in activity on maltose compared to the wild type enzyme. The catalytic efficiency (k cat/K M) was reduced fivefold but the specific activity still amounted to 63% of the wild type activity.

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Acknowledgments

This work was supported by the BMBF Innovationsinitiative Neue Länder (Unternehmen Region, InnoProfile). We would like to thank Bernd Gründig of Senslab GmbH for the helpful discussions, Christian Seidl for his help with the computer model, and Rolf Müller for his kind gift of S.cellulosum So ce56 genomic DNA.

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Authors and Affiliations

  1. Institute of Biochemistry, University of Leipzig, Deutscher Platz 5b, 04103, Leipzig, Germany

    Michael Hofer, Kathrin Bönsch, Thomas Greiner-Stöffele & Meike Ballschmiter

  2. c-LEcta GmbH, Deutscher Platz 5b, 04103, Leipzig, Germany

    Thomas Greiner-Stöffele

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  1. Michael Hofer
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  2. Kathrin Bönsch
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  3. Thomas Greiner-Stöffele
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  4. Meike Ballschmiter
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Correspondence to Meike Ballschmiter.

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Hofer, M., Bönsch, K., Greiner-Stöffele, T. et al. Characterization and Engineering of a Novel Pyrroloquinoline Quinone Dependent Glucose Dehydrogenase from Sorangium cellulosum So ce56. Mol Biotechnol 47, 253–261 (2011). https://doi.org/10.1007/s12033-010-9339-5

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