1887

Abstract

The gene cluster that encodes the periplasmic nitrate reductase from USDA110 has been isolated and characterized. encodes the catalytic subunit, and the and gene products are predicted to be a soluble dihaem and a membrane-anchored tetrahaem -type cytochrome, respectively. encodes a transmembrane protein of unknown function, and the gene product is a soluble protein which is assumed to play a role in the maturation of NapA. Western blots of the periplasmic fraction from wild-type cells grown anaerobically with nitrate revealed the presence of a protein band with a molecular size of about 90 kDa corresponding to NapA. A mutant carrying an insertion in the gene was unable to grow under nitrate-respiring conditions, lacked nitrate reductase activity, and did not show the 90 kDa protein band. Complementation of the mutant with a plasmid bearing the genes restored both nitrate-dependent anaerobic growth of the cells and nitrate reductase activity. A membrane-bound and a periplasmic -type cytochrome, with molecular masses of 25 kDa and 15 kDa, respectively, were not detected in the mutant strain incubated anaerobically with nitrate, which identifies those proteins as the NapC and the NapB components of the periplasmic nitrate reductase enzyme. These results suggest that the periplasmic nitrate reductase is the enzyme responsible for anaerobic growth of under nitrate-respiring conditions. The promoter region of the genes has been characterized by primer extension. A major transcript initiates 66·5 bp downstream of the centre of a putative FNR-like binding site.

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2003-12-01
2019-11-19
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