Nitrate reductase whole-cell assay: side effects associated with the use of benzyl viologen

Folia Microbiol (Praha). 2011 Jan;56(1):72-6. doi: 10.1007/s12223-011-0004-6. Epub 2011 Mar 17.

Abstract

A comparative examination of reduced methyl [MV·](+) and benzyl [BV·](+) viologens (as artificial electron donors for quantitative estimation of the respiratory periplasmic (Nap) and membrane-embedded (Nar) nitrate reductases) using a newly constructed nap mutant strain of Paracocccus denitrificans was done. The activity with [MV·](+) was high in whole-cell assays, confirming that this compound donates electrons to Nar. Initial rates of the more lipophilic [BV·](+) were considerably lower, which was interpreted to be caused by an inhibition of the active transport of nitrate into the cells. Anionophoric activity of [BV·](+) was detectable but too low to effectively circumvent the inhibition of nitrate transporter.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Benzyl Viologen / metabolism*
  • Electron Transport
  • Kinetics
  • Nitrate Reductase / chemistry*
  • Nitrate Reductase / genetics
  • Nitrate Reductase / metabolism
  • Nitrates / metabolism
  • Oxidation-Reduction
  • Rhodobacteraceae / chemistry
  • Rhodobacteraceae / enzymology*
  • Rhodobacteraceae / genetics
  • Single-Cell Analysis

Substances

  • Bacterial Proteins
  • Nitrates
  • Benzyl Viologen
  • Nitrate Reductase