Published: Vol 6, Iss 10, May 20, 2016 DOI: 10.21769/BioProtoc.1818 Views: 11582
Reviewed by: Valentine V TrotterGeneviève BallAnonymous reviewer(s)
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Abstract
The second step of the dissimilatory denitrification pathway in which nitrite (NO2-) is converted to nitric oxide (NO) is catalyzed by the enzyme nitrite reductase. Two distinct enzymes are found in nature that catalyze this reaction, and they contain different metal sites, either iron (Fe), in the form of heme, or copper (Cu) (Zumft, 1997). The Pseudomonas stutzeri (P. stutzeri) RCH2 strain used in this assay contains both an Fe and a Cu form of nitrite reductase. In this assay, total nitrite reductase activity can be measured in whole cells using fumarate or some other carbon source as an electron source by measuring the disappearance of nitrite over time (Thorgersen et al., 2015).
Keywords: Nitrite reductaseMaterials and Reagents
Equipment
Procedure
Representative data
Figure 1. Standard nitrite curve from 0-100 µM nitrite mixed 1:1 with Griess reagent. A. The red-pink color develops when the Griess reagent is added to the nitrite containing sample. B. A linear standard curve is shown with values ranging from 0-100 µM nitrite.
Notes
Recipes
Acknowledgments
This material by ENIGMA (Ecosystems and Networks Integrated with Genes and Molecular Assemblies) (http://enigma.lbl.gov), a Scientific Focus Area Program at Lawrence Berkeley National Laboratory, is based upon work supported by the U. S. Department of Energy, Office of Science, Office of Biological and Environmental Research, under contract number DE-AC02-05CH11231.
References
Article Information
Copyright
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Thorgersen, M. P. and Adams, M. W. W. (2016). Nitrite Reduction Assay for Whole Pseudomonas Cells. Bio-protocol 6(10): e1818. DOI: 10.21769/BioProtoc.1818.
Category
Microbiology > Microbial biochemistry > Protein
Biochemistry > Protein > Activity
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