Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences, Shanghai Key Laboratory of Agricultural Genetics and Breeding, 2901, Beidi Road, Shanghai, China; Key Laboratory for Safety Assessment (Enviornment) of Agricultural Genetically Modified Organisms, Ministry of Agriculture and Rural Affairs, Shanghai, China.
Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences, Shanghai Key Laboratory of Agricultural Genetics and Breeding, 2901, Beidi Road, Shanghai, China; Key Laboratory for Safety Assessment (Enviornment) of Agricultural Genetically Modified Organisms, Ministry of Agriculture and Rural Affairs, Shanghai, China.
Ecotoxicol Environ Saf. 2022 Sep 15;243:114016. doi: 10.1016/j.ecoenv.2022.114016. Epub 2022 Aug 24.
Nitrobenzene is widely present in industrial wastewater and soil. Biodegradation has become an ideal method to remediate organic pollutants due to its low cost, high efficiency, and absence of secondary pollution. In the present study, 10 exogenous genes that can completely degrade nitrobenzene were introduced into Escherichia coli, and their successful expression in the strain was verified by fluorescence quantitative polymerase chain reaction and proteomic analysis. The results of the degradation experiment showed that the engineered strain could completely degrade 4 mM nitrobenzene within 8 h. The formation of intermediate metabolites was detected, and the final metabolites entered the E. coli tricarboxylic acid cycle smoothly. This process was discovered by isotope tracing method. Results indicated the integrality of the degradation pathway and the complete degradation of nitrobenzene. Finally, further experiments were conducted in soil to verify its degradation ability and showed that the engineered strain could also degrade 1 mM nitrobenzene within 10 h. In this study, engineered bacteria that can completely degrade nitrobenzene have been constructed successfully. The construction of remediation-engineered bacteria by synthetic biology laid the foundation for the industrial application of biological degradation of organic pollutants.
硝基苯广泛存在于工业废水中和土壤中。由于其成本低、效率高、无二次污染,生物降解已成为修复有机污染物的理想方法。本研究将 10 种能够完全降解硝基苯的外源基因导入大肠杆菌,并通过荧光定量聚合酶链式反应和蛋白质组学分析验证了它们在菌株中的成功表达。降解实验结果表明,工程菌能在 8 h 内完全降解 4 mM 硝基苯。检测到中间代谢物的形成,最终代谢物顺利进入大肠杆菌三羧酸循环。这一过程是通过同位素示踪法发现的。结果表明降解途径的完整性和硝基苯的完全降解。最后,在土壤中进行了进一步的实验来验证其降解能力,并表明工程菌也能在 10 h 内降解 1 mM 硝基苯。本研究成功构建了能够完全降解硝基苯的工程菌。通过合成生物学构建修复工程菌为有机污染物生物降解的工业应用奠定了基础。