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Degradation of benzoate via benzoyl-coenzyme A and gentisate by Bacillus stearothermophilus PK1, and purification of gentisate 1,2-dioxygenase

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Abstract

The thermophilic Bacillus stearothermophilus PK1 utilized benzoate, 3-hydroxybenzoate, and gentisate as sole source of carbon and energy. 2- and 4-Hydroxybenzoate, 2,3- and 3,4-dihydroxybenzoate, and catechol did not support growth. Degradation of benzoate proceeded via benzoyl-coenzyme A (benzoyl-CoA) and gentisate. The inducible benzoyl-CoA ligase converted benzoate but not 3-hydroxybenzoate to its coenzyme A thioester. Gentisate 1,2-dioxygenase from B. stearothermophilus PK1 was purified to homogeneity. The enzyme is presumed to be a homohexamer with a subunit molecular mass of 40 kDa. It showed maximal activity at 65–70°C. After incubation for 80 min at 65°C, 50% of the original activity was lost. Gentisate 1,2-dioxygenase activity from strain PK1 was strictly dependent on exogenously added Fe2+, and it was inhibited by metal-chelating agents, indicating an essential role of Fe2+ in catalysis.

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Authors and Affiliations

  1. Institut für Mikrobiologie (250), Universität Hohenheim, D-70593, Stuttgart, Germany

    P. Kiemer, B. Tshisuaka, S. Fetzner & F. Lingens

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  1. P. Kiemer
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  2. B. Tshisuaka
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  3. S. Fetzner
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  4. F. Lingens
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Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday

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Kiemer, P., Tshisuaka, B., Fetzner, S. et al. Degradation of benzoate via benzoyl-coenzyme A and gentisate by Bacillus stearothermophilus PK1, and purification of gentisate 1,2-dioxygenase. Biol Fertil Soils 23, 307–313 (1996). https://doi.org/10.1007/BF00335959

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  • DOI: https://doi.org/10.1007/BF00335959

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