A novel pathway of aerobic benzoate catabolism in the bacteria Azoarcus evansii and Bacillus stearothermophilus

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-(13)C(6)]benzoyl-CoA as substrate, cis-3,4-2,3,4,5,6-(13)C(5)]dehydroadipyl-CoA, trans-2,3-2,3,4,5,6-(13)C(5)]dehydroadipyl-CoA, the 3,6-lactone of 3-2,3,4,5,6-(13)C(5)]hydroxyadipyl-CoA, and 3-2,3,4,5,6-(13)C(5)]hydroxyadipyl-CoA were identified as products by NMR spectroscopy. A protein mixture of A. evansii transformed ring-(13)C(6)]benzoyl-CoA in an NADPH- and oxygen-dependent reaction into 6-2,3,4,5,6-(13)C(5)]hydroxy-3-hexenoyl-CoA. The data suggest a novel aerobic pathway of benzoate catabolism via CoA intermediates leading to beta-ketoadipyl-CoA, an intermediate of the known beta-ketoadipate pathway.