Reductive dehalogenase structure suggests a mechanism for B12-dependent dehalogenation
Open Access
- 19 October 2014
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature
- Vol. 517 (7535), 513-516
- https://doi.org/10.1038/nature13901
Abstract
X-ray crystallography and EPR spectroscopy are used to characterize a soluble, oxygen-tolerant reductive dehalogenase from Nitratireductor pacificus pht-3B; the data suggest that the cobalt in the cobalamin cofactor ligates the halogen atom of the substrate, directly abstracting the halogen atom via an oxidative addition. Reductive dehalogenases are cobalamin-dependent enzymes that catalyse the removal of a halogen atom from organohalides, organic molecules that contain one or more halogen atoms. A large proportion of environmental pollutants are organohalides and reductive dehalogenases are responsible for the biological dehalogenation of these compounds, including polychlorinated biphenyls. In this manuscript, the authors used X-ray crystallography and EPR spectroscopy to characterize a soluble, oxygen-tolerant reductive dehalogenase, pht-3B, from Nitratireductor pacificus. Their data suggest that the cobalt in the cobalamin cofactor forms a covalent bond with the halogen atom of the substrate, directly abstracting the halogen atom via an oxidative addition. This mechanism is fundamentally different from the mechanisms of other cobalamin-containing enzymes. These findings will be of relevance to future use of reductive dehalogenases in bioremediation and biocatalysis. Organohalide chemistry underpins many industrial and agricultural processes, and a large proportion of environmental pollutants are organohalides1. Nevertheless, organohalide chemistry is not exclusively of anthropogenic origin, with natural abiotic and biological processes contributing to the global halide cycle2,3. Reductive dehalogenases are responsible for biological dehalogenation in organohalide respiring bacteria4,5, with substrates including polychlorinated biphenyls or dioxins6,7. Reductive dehalogenases form a distinct subfamily of cobalamin (B12)-dependent enzymes that are usually membrane associated and oxygen sensitive, hindering detailed studies8,9,10,11,12. Here we report the characterization of a soluble, oxygen-tolerant reductive dehalogenase and, by combining structure determination with EPR (electron paramagnetic resonance) spectroscopy and simulation, show that a direct interaction between the cobalamin cobalt and the substrate halogen underpins catalysis. In contrast to the carbon–cobalt bond chemistry catalysed by the other cobalamin-dependent subfamilies13, we propose that reductive dehalogenases achieve reduction of the organohalide substrate via halogen–cobalt bond formation. This presents a new model in both organohalide and cobalamin (bio)chemistry that will guide future exploitation of these enzymes in bioremediation or biocatalysis.Keywords
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