Gene cloning, expression and performance validation of nitric oxide dismutase

• Published in: The Science of the total environment Vol. 935; p. 173455
• Main Authors: Bai, Mengwei, He, Jiamei, Zheng, Fengzhen, Lv, Sini, Wang, Zeyu, Hrynsphan, Dzmitry, Savitskaya, Tatsiana, Chen, Jun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 20.07.2024
• Subjects: biodegradation; Biodegradation, Environmental; carbon dioxide; Cloning, Molecular; denitrification; environment; Escherichia coli; Escherichia coli - genetics; genes; N2O; New pathway; nitric oxide; nitrogen; Nitrous Oxide; Nod gene; plasmids; pollution; Recombinant engineering bacteria; Sewage - microbiology; sludge; N2O; New pathway; Nod gene; Recombinant engineering bacteria; NO
• ISSN: 0048-9697; 1879-1026; 1879-1026
• DOI: 10.1016/j.scitotenv.2024.173455

Nitrous oxide (N2O) is a significant contributor to global warming and possesses an ozone-depleting impact nearly 298 times that of CO2. To reduce N2O emissions, the newly-discovered nod gene which can directly convert NO into N2 and O2 was successfully cloned from the anaerobic denitrification sludge. The recombinant plasmid containing the nod gene was built, and the expression of nod gene in Escherichia coli was determined, leading to the construction of recombinant engineering bacteria. Results showed that the recombinant engineering bacteria E. coli BL21 (DE3)-pET28a-nod could autonomously degrade NO, with a degradation rate of 72 % within 48 h, and could produce 2479.72 ppm of N2 and 75.12 mL of O2. The cumulative O2 production of the sludge sample and recombinant E. coli within 8 h was 1.75 mL and 8.45 mL, respectively. The cumulative O2 production of recombinant E. coli was at least 4.82 times higher than that of the sludge sample. The investigation proposed a new biodegradation pathway for nitrogen pollution. [Display omitted] •The newly-discovered nod gene which can directly convert NO into N2 and O2 was successfully cloned.•The recombinant engineering bacteria were constructed and the nod gene was expressed in Escherichia coli.•The E. coli BL21 (DE3)-pET28a-nod has the good performance for O2 production.