School of Biology and Biological Engineering, South China University of Technology, Higher Education Mega Center, Guangzhou, 510006, China.
Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, 510006, China.
Appl Microbiol Biotechnol. 2022 Sep;106(18):6317-6333. doi: 10.1007/s00253-022-12136-1. Epub 2022 Aug 27.
Recombinant luminescent Escherichia coli strains could be used to detect the toxicity of pure or mixed contaminants as a light-off sensor. In this work, the lux operon of Photobacterium phosphoreum T3 was identified for the first time. Recombinant luminescent E. coli strains were constructed via expressing the lux operons of P. phosphoreum T3 and Vibrio qinghaiensis Q67 in E. coli MG1655, and the optimal protectant containing 10% (w/v) trehalose and 4% sucrose was used to prepare the freeze-dried recombinant luminescent E. coli cells. Then, these freeze-dried E. coli cells were subjected to acute toxicity detection. The results showed that luminescent E. coli strains displayed sensitive toxic responses to BPA, nFeO, Cd, Pb, As, and Hg, for example, the EC values of BPA and nFeO to luminescent E. coli strains ranged from 1.54 to 50.19 mg/l and 17.50 to 21.52 mg/l, respectively. Indeed, luminescent E. coli strains exhibited more sensitive responses to Cd, Pb, and Hg than the natural strain Q67. The results suggested that recombinant luminescent E. coli strains could be used for the detection of acute toxicity. Furthermore, the combined toxicities of BPA and nFeO, Hg, and Pb were measured, and the joint effects of these mixtures were evaluated with luminescent E. coli. The results indicated that the joint effects of BPA and nFeO suggested to be synergistic or additive to luminescent E. coli, while the joint effects of heavy metals and nFeO exhibited additivities. The cellular endocytosis for FeO nanoparticles was not observed, which could explain the additive instead of synergistic effects between heavy metals and nFeO. KEY POINTS: • Sequence of the lux operon from P. phosphoreum T3 was reported for the first time. • Recombinant luminescent E. coli was more sensitive to Cd, Pb, and Hg than Q67. • Joint effects of BPA and nFeO were synergistic or additive to luminescent E. coli.
重组发光大肠杆菌菌株可作为光启动传感器用于检测纯污染物或混合污染物的毒性。在这项工作中,首次鉴定出了发光异养菌 T3 的 lux 操纵子。通过在大肠杆菌 MG1655 中表达发光异养菌 T3 和青海弧菌 Q67 的 lux 操纵子,构建了重组发光大肠杆菌菌株,并使用含有 10%(w/v)海藻糖和 4%蔗糖的最佳保护剂来制备冻干的重组发光大肠杆菌细胞。然后,对这些冻干的大肠杆菌细胞进行急性毒性检测。结果表明,发光大肠杆菌菌株对 BPA、nFeO、Cd、Pb、As 和 Hg 等物质表现出敏感的毒性反应,例如 BPA 和 nFeO 对发光大肠杆菌菌株的 EC 值范围分别为 1.54 至 50.19mg/L 和 17.50 至 21.52mg/L。实际上,发光大肠杆菌菌株对 Cd、Pb 和 Hg 的反应比自然菌株 Q67 更为敏感。结果表明,重组发光大肠杆菌菌株可用于急性毒性检测。此外,还测量了 BPA 和 nFeO、Hg 和 Pb 的联合毒性,并使用发光大肠杆菌评估了这些混合物的联合作用。结果表明,BPA 和 nFeO 的联合作用对发光大肠杆菌表现出协同或相加作用,而重金属和 nFeO 的联合作用则表现出相加作用。未观察到 FeO 纳米颗粒的细胞内内吞作用,这可以解释重金属和 nFeO 之间的相加作用而不是协同作用。关键点:
首次报道了发光异养菌 T3 的 lux 操纵子序列。
与 Q67 相比,重组发光大肠杆菌对 Cd、Pb 和 Hg 更为敏感。
BPA 和 nFeO 的联合作用对发光大肠杆菌表现出协同或相加作用。