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Isolation, expression, and biochemical characterization: nitrite reductase from Bacillus cereus LJ01
Yan-Yan Huang1, Ming-Hua Liang1, Shan Zhao1
1School of Food Science and Engineering, South China University of Technology 381 Wushan Road Guangzhou Guangdong 510640 People's Republic of China liudm@scut.edu.cn.
RSC Advances
|May 6, 2022
Summary
This study identifies a novel nitrite reductase (LJ01-NiR) from Bacillus cereus LJ01, demonstrating its high efficiency in degrading nitrite pollution. The enzyme
Area of Science:
- Environmental microbiology
- Biotechnology
- Enzyme kinetics
Background:
- Nitrite is a common environmental pollutant requiring effective bioremediation strategies.
- Understanding the enzymes involved in nitrite degradation is crucial for developing these strategies.
Purpose of the Study:
- To isolate and characterize a nitrite-degrading enzyme from Bacillus cereus LJ01.
- To evaluate the potential of this enzyme for environmental bioremediation.
Main Methods:
- Isolation and purification of nitrite-degrading active substance (ASDN) from B. cereus LJ01.
- Cloning and overexpression of the nitrite reductase gene (LJ01-NiR) in E. coli.
- Enzyme activity assays, kinetic analysis, and structural analysis of LJ01-NiR.
Main Results:
- Bacillus cereus LJ01 effectively degraded nitrite, with the isolated nitrite reductase (LJ01-NiR) showing high specific activity (4004.89 U mg-1).
- Recombinant LJ01-NiR exhibited broad activity under various temperature, pH, and metal ion conditions.
- Kinetic parameters (Km = 1.38 mM, Vmax = 2.00 μmol g-1 min-1) and structural analysis indicated it is a ferredoxin-dependent nitrite reductase.
Conclusions:
- LJ01-NiR possesses high nitrite-degrading capability, making it a promising candidate for environmental pollution treatment.
- The enzyme's stability and activity profile support its potential application in bioremediation technologies.
