Mora-Garduño Josué Daniel, Tamayo-Nuñez Jessica, Padilla-Vaca Felipe, Ramírez-Montiel Fátima Berenice, Rangel-Serrano Ángeles, Santos-Escobar Fernando, Gutiérrez-Corona Félix, Páramo-Pérez Itzel, Anaya-Velázquez Fernando, García-Contreras Rodolfo, Vargas-Maya Naurú Idalia, Franco Bernardo
Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta S/N, 36050, Guanajuato, Gto, Mexico.
Departamento de Microbiología y Parasitología, Facultad de Medicina, UNAM, Mexico City, Mexico.
AMB Express. 2022 Jan 6;12(1):2. doi: 10.1186/s13568-021-01342-1.
The presence of pollutants in soil and water has given rise to diverse analytical and biological approaches to detect and measure contaminants in the environment. Using bacterial cells as reporter strains represents an advantage for detecting pollutants present in soil or water samples. Here, an Escherichia coli reporter strain expressing a chromoprotein capable of interacting with soil or water samples and responding to DNA damaging compounds is validated. The reporter strain generates a qualitative signal and is based on the expression of the coral chromoprotein AmilCP under the control of the recA promoter. This strain can be used simply by applying soil or water samples directly and rendering activation upon DNA damage. This reporter strain responds to agents that damage DNA (with an apparent detection limit of 1 µg of mitomycin C) without observable response to membrane integrity damage, protein folding or oxidative stress generating agents, in the latter case, DNA damage was observed. The developed reporter strain reported here is effective for the detection of DNA damaging agents present in soils samples. In a proof-of-concept analysis using soil containing chromium, showing activation at 15.56 mg/L of Cr(VI) present in soil and leached samples and is consistent with Cr(III) toxicity at high concentrations (130 µg). Our findings suggest that chromogenic reporter strains can be applied for simple screening, thus reducing the number of samples requiring analytical techniques.
土壤和水中污染物的存在催生了多种用于检测和测量环境中污染物的分析和生物学方法。使用细菌细胞作为报告菌株在检测土壤或水样中的污染物方面具有优势。在此,一种表达能够与土壤或水样相互作用并对DNA损伤化合物作出反应的显色蛋白的大肠杆菌报告菌株得到了验证。该报告菌株产生定性信号,基于recA启动子控制下珊瑚显色蛋白AmilCP的表达。通过直接应用土壤或水样并在DNA损伤时实现激活,即可简单地使用该菌株。该报告菌株对损伤DNA的试剂有反应(丝裂霉素C的表观检测限为1微克),而对膜完整性损伤、蛋白质折叠或氧化应激产生试剂无明显反应,在后一种情况下,观察到了DNA损伤。本文报道的开发的报告菌株可有效检测土壤样品中存在的DNA损伤剂。在一项使用含铬土壤的概念验证分析中,该菌株在土壤和浸出样品中15.56毫克/升的六价铬存在时显示出激活,并且与高浓度(130微克)三价铬的毒性一致。我们的研究结果表明,显色报告菌株可用于简单筛选,从而减少需要分析技术的样品数量。