| Autor: |
Halliwell T; Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK., Fisher K; Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK., Payne KAP; Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK.; Future Biomanufacturing Research Hub (FutureBRH), Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK., Rigby SEJ; Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK., Leys D; Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK. |
| Abstrakt: |
Reductive dehalogenases are responsible for the reductive cleavage of carbon-halogen bonds during organohalide respiration. A variety of mechanisms have been proposed for these cobalamin and [4Fe-4S] containing enzymes, including organocobalt, radical, or cobalt-halide adduct based catalysis. The latter was proposed for the oxygen-tolerant Nitratireductor pacificus pht-3B cataboli c reductive dehalogenase (NpRdhA). Here, we present the first substrate bound NpRdhA crystal structures, confirming a direct cobalt-halogen interaction is established and providing a rationale for substrate preference. Product formation is observed in crystallo due to X-ray photoreduction. Protein engineering enables rational alteration of substrate preference, providing a future blue print for the application of this and related enzymes in bioremediation. |
