Related Experiment Video
Updated: May 21, 2026

Measurement of the Potential Rates of Dissimilatory Nitrate Reduction to Ammonium Based on 14NH4+/15NH4+ Analyses via Sequential Conversion to N2O
Published on: October 7, 2020
Expression, and molecular and enzymatic characterization of Cu-containing nitrite reductase from a marine
Keitaro Kondo1, Katsuhiko Yoshimatsu, Taketomo Fujiwara
1Department of Biological Science, Graduate School of Science, Shizuoka University, 836 Oh-ya, Suruga-ku, Shizuoka 422–8529, Japan.
Abstract:
Ammonia-oxidizing bacteria (AOB) remove intracellular nitrite to prevent its toxicity by a nitrifier denitrification pathway involving two denitrifying enzymes, nitrite reductase and nitric oxide reductase. Here, a Cu-containing nitrite reductase from Nitrosococcus oceani strain NS58, a gammaproteobacterial marine AOB, was expressed in Escherichia coli and purified to homogeneity. Sequence homology analysis indicated that the nitrite reductase from N. oceani was phylogenetically closer to its counterparts from denitrifying bacteria than that of the betaproteobacterium Nitrosomonas europaea. The recombinant enzyme was a homotrimer of a 32 kDa subunit molecule. The enzyme was green in the oxidized state with absorption peaks at 455 nm and 575 nm. EPR spectroscopy indicated the presence of type 2 Cu. Molecular activities and the affinity constant for the nitrite were determined to be 1.6×10(3) s(-1) and 52 μM, respectively.
More Related Videos
08:23Chemiluminescence-based Assays for Detection of Nitric Oxide and its Derivatives from Autoxidation and Nitrosated Compounds
Published on: February 16, 2022
08:32Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
Published on: March 16, 2017
Related Concept Videos
Inorganic Nitrogen Assimilation
Metabolism of Chemolithotrophs
The Nitrogen Cycle
Deep Sea Microbial Ecology
2° Amines to N-Nitrosamines: Reaction with NaNO2
Sulfur Assimilation