Department of Chemistry and Biochemistry, Indiana University, South Bend, IN 46615, USA.
Protein Eng Des Sel. 2020 Sep 14;33. doi: 10.1093/protein/gzaa021.
Glyphosate, the active ingredient in RoundUp, is the most widely used herbicide on the globe, and has recently been linked to an increased risk in non-Hodgkin's lymphoma in exposed individuals. Therefore, detection and monitoring of glyphosate levels in water and soil is important for public safety. Here, we describe a biosensor for glyphosate based on an engineered Escherichia coli phosphonate-binding protein (PhnD). Mutations in the binding pocket were introduced to convert PhnD into a glyphosate-binding protein. A fluorescence group attached near the hinge of the protein was added to monitor binding of glyphosate and to determine its concentration in unknown samples. The resulting engineered biosensor can detect glyphosate in tap water and in soil samples treated with the herbicide at submicromolar concentrations, well below the limit for drinking water in the USA. Incorporating this biosensor in a device would allow rapid and continuous monitoring of glyphosate in water and soil samples.
草甘膦是农达除草剂的有效成分,是全球使用最广泛的除草剂,最近有研究表明,接触草甘膦会增加非霍奇金淋巴瘤的风险。因此,检测和监测水和土壤中的草甘膦水平对于公共安全非常重要。在这里,我们描述了一种基于工程化大肠杆菌膦酸盐结合蛋白(PhnD)的草甘膦生物传感器。在结合口袋中引入突变,将 PhnD 转化为草甘膦结合蛋白。在蛋白质铰链附近添加一个荧光基团,用于监测草甘膦的结合,并确定未知样品中的浓度。由此产生的工程生物传感器可以检测自来水中和经除草剂处理的土壤样品中的草甘膦,浓度低至亚微摩尔,远低于美国饮用水的限量。将该生物传感器整合到设备中,可以实现对水和土壤样品中草甘膦的快速连续监测。