Rapid field testing of mercury pollution by designed fluorescent biosensor and its cells-alginate hydrogel-based paper assay

With increasing industrial activities,mercury has been largely discharged into environment and caused serious environmental problems.The growing level of mercury pollution has become a huge threat to human health due to its significant biotoxicity.Therefore,the simple and fast means for on-site monitoring discharged mercury pollution are highly necessary to protect human beings from its pernicious effects in time.Herein,a"turn off"fluorescent biosensor(mCherry L199 C)for sensing Hg2+was successfully designed based on direct modification of the chromophore environment of fluorescent protein mCherry.For rapid screening and characterization,the designed variant of mCherry(mCherry L199 C)was directly expressed on outer-membrane of Escherichia coli cells by cell surface display technique.The fluorescent biosensor was characterized to have favorable response to Hg2+at micromole level among other metal ions and over a broad pH range.Further,the cells of the fluorescent biosensor were encapsulated in alginate hydrogel to develop the cells-alginate hydrogel-based paper.The cells-alginate hydrogel-based paper could detect mercury pollution in 5 min with simple operation process and inexpensive equipment,and it could keep fluorescence and activity stable at 4℃ for 24 hr,which would be a high-throughput screening tool in preliminarily reporting the presence of mercury pollution in natural setting.