Expression and characterization of the assimilatory NADH-nitrite reductase from the phototrophic bacterium Rhodobacter capsulatus E1F1

Arch Microbiol. 2006 Oct;186(4):339-44. doi: 10.1007/s00203-006-0149-x. Epub 2006 Aug 3.

Abstract

A nas gene region from Rhodobacter capsulatus E1F1 containing the putative nasB gene for nitrite reductase was previously cloned. The recombinant His(6)-NasB protein overproduced in E. coli showed nitrite reductase activity in vitro with both reduced methyl viologen and NADH as electron donors. The apparent K ( m ) values for nitrite and NADH were 0.5 mM and 20 microM, respectively, at the pH and temperature optima (pH 9 and 30 degrees C). The optical spectrum showed features that indicate the presence of FAD, iron-sulfur cluster and siroheme as prosthetic groups, and nitrite reductase activity was inhibited by sulfide and iron reagents. These results indicate that the phototrophic bacterium R. capsulatus E1F1 possesses an assimilatory NADH-nitrite reductase similar to that described in non-phototrophic organisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Flavin-Adenine Dinucleotide / chemistry
  • Heme / analogs & derivatives
  • Heme / chemistry
  • NAD / metabolism*
  • Nitrates / metabolism
  • Nitrite Reductases / chemistry
  • Nitrite Reductases / genetics
  • Nitrite Reductases / metabolism*
  • Paraquat / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Rhodobacter capsulatus / enzymology*
  • Rhodobacter capsulatus / genetics
  • Rhodobacter capsulatus / growth & development

Substances

  • Nitrates
  • Recombinant Proteins
  • NAD
  • Flavin-Adenine Dinucleotide
  • Heme
  • siroheme
  • Nitrite Reductases
  • Paraquat