Abstract
Dissimilatory nitrite reductase, a key enzyme in the denitrification pathway, catalyzes the reduction of nitrite to NO. Bioinformatic analysis showed that the genome of a novel nitrite-degrading haloarchaeon Halorussus sp. YCN54 possessed a gene encoding the Cu-containing dissimilatory nitrite reductase (NirKHrs). NirKHrs was heterologously expressed and purified. Protein sequencing indicated that two isoforms of NirKHrs monomer were produced intracellularly. UV–vis spectrum of the purified NirKHrs showed that it belonged to the blue NirK group. NirKHrs showed optimum activity at 4.5 M NaCl, 55 ℃ and pH 7.0, representing a halophilic, slightly thermophilic and neutral enzyme. It exhibited high stability at 30–50 ℃. NirKHrs activity was strongly inhibited by the copper chelating agent due to removal of copper. NirKHrs activity was activated by Mn2+ and Sr2+. It displayed good tolerance to some high polarity organic solvents and nonionic surfactants, such as glycerol, DMSO, DMF and tween-20. Na2S2O4 was an effective electron donor to NirKHrs. The Km and Vmax values of purified NirKHrs for nitrite were 3.2 mM and 477.2 U/mg, respectively, indicating its high activity. These results indicated that NirKHrs may have potential applications for nitrite degradation in high-salt industries, such as salted food and saline wastewater treatment.
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Abbreviations
- MV:
-
Methyl viologen
- NirK:
-
Cu-containing nitrite reductases
- NirS:
-
Cytochrome-c-dependent nitrite reductases
- DMSO:
-
Dimethyl sulfoxide
- DMF:
-
N,N-dimethylformamide
- DDC:
-
Diethyldithiocarbamate
- Tat:
-
Twin-arginine translocation
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Acknowledgements
This work was financially supported by grants from National Natural Science Foundation of China (No. 31600002 and No. 31770005) and the Advanced Talent Foundation of Jiangsu University (No. 15JDG062).
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Hou, J., Yang, XY., Xu, Q. et al. Characterization of a novel Cu-containing dissimilatory nitrite reductase from the haloarchaeon Halorussus sp. YCN54. Extremophiles 24, 403–411 (2020). https://doi.org/10.1007/s00792-020-01164-2
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DOI: https://doi.org/10.1007/s00792-020-01164-2