title: A cluster of bacterial genes for anaerobic benzene ring biodegradation.
abstract: A reductive benzoate pathway is the central conduit for the anaerobic biodegradation of aromatic pollutants and lignin monomers. Benzene ring reduction requires a large input of energy and this metabolic capability has, so far, been reported only in bacteria. To determine the molecular basis for this environmentally important process, we cloned and analyzed genes required for the anaerobic degradation of benzoate and related compounds from the phototrophic bacterium, Rhodopseudomonas palustris. A cluster of 24 genes was identified that includes twelve genes likely to be involved in anaerobic benzoate degradation and additional genes that convert the related compounds 4-hydroxybenzoate and cyclohexanecarboxylate to benzoyl-CoA. Genes encoding benzoyl-CoA reductase, a novel enzyme able to overcome the resonance stability of the aromatic ring, were identified by directed mutagenesis. The gene encoding the ring-cleavage enzyme, 2-ketocyclohexanecarboxyl-CoA hydrolase, was identified by assaying the enzymatic activity of the protein expressed in Escherichia coli. Physiological data and DNA sequence analyses indicate that the benzoate pathway consists of unusual enzymes for ring reduction and cleavage interposed among enzymes homologous to those catalyzing fatty acid degradation. The cloned genes should be useful as probes to identify benzoate degradation genes from other metabolically distinct groups of anaerobic bacteria, such as denitrifying bacteria and sulfate-reducing bacteria.
authors.affiliation: Department of Microbiology, and Center for Biocatalysis and Bioprocessing, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: P G
authors.initials: PG
authors.lastname: Egland
authors.forename: D A
authors.initials: DA
authors.lastname: Pelletier
authors.forename: M
authors.initials: M
authors.lastname: Dispensa
authors.forename: J
authors.initials: J
authors.lastname: Gibson
authors.forename: C S
authors.initials: CS
authors.lastname: Harwood
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics*; D001554:Benzene / Q000378:metabolism*; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D004926:Escherichia coli; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012241:Rhodopseudomonas / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D013869:Thiolester Hydrolases / Q000096:biosynthesis / Q000737:chemistry; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; C106551:2-ketocyclohexanecarboxyl-coenzyme A hydrolase; D013869:Thiolester Hydrolases; D001616:beta-Galactosidase; D001554:Benzene