Abstract
Rhodobacter capsulatus E1F1 is a phototrophic bacterium that assimilates nitrate under light-anaerobic growth conditions. We have recently cloned a 18-kb nas gene region with 14 genes involved in the nitrate assimilation process, including the hcp gene encoding a hybrid cluster protein, and the putative structural genes coding for the catalytic subunits of nitrate reductase (nasA) and nitrite reductase nasB). Recombinant His6-tagged forms of R. capsulatus Hcp and NasB proteins have been overexpressed in E. coli, purified by affinity chromatography and biochemically characterized. These results indicate that NasB is an assimilatory NADH-nitrite reductase with siroheme, FAD and [4Fe-4S], and that the Hcp protein is a hydroxylamine reductase probably involved in detoxification and assimilation of hydroxy lamine.
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