Journal
BIO-PROTOCOL
Volume 11, Issue 14, Pages -Publisher
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.4098
Keywords
Mycolicibacterium smegmatis; Assimilatory nitrate reductase; In vitro nitrate reductase activity assay; NADPH; NADH; Methyl viologen
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Funding
- National Natural Science Foundation of China [31430004, 31830002, 81671988, 91751000, 91951000]
- Research Unit Fund of Li Ka Shing Institute of Health Sciences [7103506]
- Hong Kong Health and Medical Research Fund [12110622]
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Nitrate serves as a major inorganic nitrogen source for microorganisms, with many bacterial and archaeal lineages expressing cytoplasmic assimilatory nitrate reductase (NAS) to catalyze the reduction of nitrate to nitrite. This study presents a detailed protocol for measuring in vitro nitrate reductase (NaR) activity of NAS enzymes from Mycolicibacterium smegmatis crude extract using both physiological and non-physiological electron donors.
Nitrate is one of the major inorganic nitrogen sources for microorganisms. Many bacterial and archaeal lineages can express cytoplasmic assimilatory nitrate reductase (NAS), which catalyzes the rate-limiting reduction of nitrate to nitrite in the nitrate assimilation pathway. Here, we present a detailed protocol for measuring in vitro nitrate reductase (NaR) activity of NAS enzymes from Mycolicibacterium smegmatis crude extract using both physiological and non-physiological electron donors.
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