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Purification and properties of D-gluconate dehydratase fromAchromobacter

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Abstract

d-Gluconate dehydratase fromAchromobacter, grown ond-gluconate, was purified 100-fold by a procedure involving ammonium sulfate fractionation and preparative acrylamide gel electrophoresis. The purified enzyme appeared to be homogeneous by disc gel electrophoresis. It is an inducible enzyme with an optimal activity in the pH region 8.4–8.8, a Km value of 2.08 × 10−2 m ford-gluconate and a molecular weight of 270,000 ± 25,000. Only C5 and C6 aldonic acids possessing al-threo configuration at C2 and C3 are dehydrated. The dehydration products ofd-gluconate,d-xylonate,d-galactonate,d-fuconate andl-arabonate were identified as 2-keto-3-deoxy compounds by specific colour reactions and thin layer chromatography. Onemm Mg+ + is a powerful activator, 0.1 mm Mn+ + activates poorly and EDTA inhibits. Glutatione, dithiothreitol and mercaptoethanol had no effect, althoughp-chloromercuribenzoate (0.01 mm) decreased enzyme activity.

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Author notes

  1. Junko Khan-Matsubara

    Present address: School of Health Sciences, Faculty of Medicine, University of Tokyo, Tokyo, Japan

Authors and Affiliations

  1. Laboratory for Microbiology and Microbial Genetics, Faculty of Sciences, State University, Gent, Belgium

    K. Kersters, Junko Khan-Matsubara, L. Nelen & J. De Ley

Authors
  1. K. Kersters
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  2. Junko Khan-Matsubara
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  3. L. Nelen
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  4. J. De Ley
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Additional information

We wish to thank Mr D. Dewettinck for skilful technical assistance. The senior author (J.D.L.) is indebted to the Fonds voor Kollektief en Fundamenteel Onderzoek (Belgium) for research and personnel grants. J.K.-M. is indebted to the Belgian government for a travel and study grant.

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Kersters, K., Khan-Matsubara, J., Nelen, L. et al. Purification and properties of D-gluconate dehydratase fromAchromobacter . Antonie van Leeuwenhoek 37, 233–246 (1971). https://doi.org/10.1007/BF02218486

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