Gescher, JohannesJohannesGescherIsmail, WaelWaelIsmailÖlgeschläger, EllenEllenÖlgeschlägerEisenreich, WolfgangWolfgangEisenreichWörth, JürgenJürgenWörthFuchs, GeorgGeorgFuchs2023-02-102023-02-102006-04-15Journal of Bacteriology 188 (8): 2919-2927 (2006-04-15)http://hdl.handle.net/11420/14790Benzoate, a strategic intermediate in aerobic aromatic metabolism, is metabolized in various bacteria via an unorthodox pathway. The intermediates of this pathway are coenzyme A (CoA) thioesters throughout, and ring cleavage is nonoxygenolytic. The fate of the ring cleavage product 3,4-deliydroadipyl-CoA semialdehyde was studied in the β-proteobacterium Azoarcws evansii. Cell extracts contained a benzoate-induced, NADP+-specific aldehyde dehydrogenase, which oxidized this intermediate. A postulated putative long-chain aldehyde dehydrogenase gene, which might encode this new enzyme, is located on a cluster of genes encoding enzymes and a transport system required for aerobic benzoate oxidation. The gene was expressed in Escherichia coli, and the maltose-binding protein-tagged enzyme was purified and studied. It is a homodimer composed of 54 kDa (without tag) subunits and was confirmed to be the desired 3,4-dehydroadipyl-CoA semialdehyde dehydrogenase. The reaction product was identified by nuclear magnetic resonance spectroscopy as the corresponding acid 3,4-dehydroadipyl-CoA. Hence, the intermediates of aerobic benzoyl-CoA catabolic pathway recognized so far are benzoyl-CoA; 2,3-dihydro-2,3- dihydroxybenzoyl-CoA; 3,4-dehydroadipyl-CoA semialdehyde plus formate; and 3,4-dehydroadipyl-CoA. The further metabolism is thought to lead to 3-oxoadipyl-CoA, the intermediate at which the conventional and the unorthodox pathways merge.en0021-91932006829192927American Society for MicrobiologyBiowissenschaften, BiologieJournal Article10.1128/JB.188.8.2919-2927.200616585753Journal Article