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Effects of site-specific mutations on the enzymatic properties of a sialidase fromClostridium perfringens

Glycoconjugate Journal volume 9, pages 235–240 (1992)Cite this article

Abstract

Three site-specific mutations were performed in two regions of a sialidase gene fromClostridium perfringens which are known to be conserved in bacterial sialidases. The mutant enzymes were expressed inEscherichia coli and, when measured with MU-Neu5Ac as substrate, exhibited variations in enzymatic properties compared with the wild-type enzyme. The conservative substitution of Arg 37 by Lys, located in a short conserved region upstream from the four repeated sequences common in bacterial sialidase genes, was of special interest, asK M andV max, as well asK i measured with Neu5Ac2en, were dramatically changed. These data suggest that this residue may be involved in substrate binding. In addition to its low activity, this mutant enzyme has a lower temperature optimum and is active over a more limited pH range. This mutation also prevents the binding of an antibody able to inhibit the wild-type sialidase. The other mutations, located in one of the consensus sequences, were of lower influence on enzyme activity and recognition by antibodies.

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Affiliations

  1. Biochemisches Institut, Christian-Albrechts-Universität, Olshausenstr. 40, W-2300, Kiel, Germany

    Telse Roggentin, Reinhard G. Kleineidam, Roland Schauer & Peter Roggentin

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  1. Telse Roggentin
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  2. Reinhard G. Kleineidam
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  3. Roland Schauer
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  4. Peter Roggentin
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Roggentin, T., Kleineidam, R.G., Schauer, R. et al. Effects of site-specific mutations on the enzymatic properties of a sialidase fromClostridium perfringens . Glycoconjugate J 9, 235–240 (1992). https://doi.org/10.1007/BF00731135

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  • DOI: https://doi.org/10.1007/BF00731135

Keywords

  • sialidase (neuraminidase)
  • site-specific mutations
  • conserved sequences
  • enzymatic properties
  • Clostridium perfringens