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J. Biol. Chem., Vol. 276, Issue 27, 24997-25004, July 6, 2001

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A Novel Pathway of Aerobic Benzoate Catabolism in the Bacteria Azoarcus evansii and Bacillus stearothermophilus^*

Annette Zaar, Wolfgang Eisenreich^§¶, Adelbert Bacher^§, and Georg Fuchs

From the  Institut für Biologie II, Mikrobiologie, Universität Freiburg, Schänzlestraße 1, D-79104 Freiburg, Germany and the ^§ Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstraße 4, D-85747 Garching, Germany

The aerobic catabolism of benzoate was studied in the Gram-negative proteobacterium Azoarcus evansii and in the Gram-positive bacterium Bacillus stearothermophilus. In contrast to earlier proposals, benzoate was not converted into hydroxybenzoate or gentisate. Rather, benzoyl-CoA was a product of benzoate catabolism in both microbial species under aerobic conditions in vivo. Benzoyl-CoA was converted into various CoA thioesters by cell extracts of both species in oxygen- and NADPH-dependent reactions. Using [ring-¹³C₆]benzoyl-CoA as substrate, cis-3,4-[2,3,4,5,6-¹³C₅]dehydroadipyl-CoA, trans-2,3-[2,3,4,5,6-¹³C₅]dehydroadipyl-CoA, the 3,6-lactone of 3-[2,3,4,5,6-¹³C₅]hydroxyadipyl-CoA, and 3-[2,3,4,5,6-¹³C₅]hydroxyadipyl-CoA were identified as products by NMR spectroscopy. A protein mixture of A. evansii transformed [ring-¹³C₆]benzoyl-CoA in an NADPH- and oxygen-dependent reaction into 6-[2,3,4,5,6-¹³C₅]hydroxy-3-hexenoyl-CoA. The data suggest a novel aerobic pathway of benzoate catabolism via CoA intermediates leading to -ketoadipyl-CoA, an intermediate of the known -ketoadipate pathway.

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