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Construction of WCB-11:A novel phiYFP arsenic-resistant whole-cell biosensor

Journal of Environmental Sciences 2010年第9期22卷 1469-1474页

作者： Qing Hu ,Li Li,Yanjuan Wang,Wenjuan Zhao,Hongyan Qi,Guoqiang Zhuang Department of Environmental Bio-Technology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China. Department of Environmental Bio-Technology Research Center for Eco-Environmental Sciences Chinese Academy of Sciences Beijing 100085 Chin

The prediction and assessment of environmental pollution by arsenic are important preconditions of advocating environmental protection and human health risk assessment. A yellow fluorescent protein-based whole-cell biosensor for the detection of arsenite and arsenate was constructed and tested. An arsenic-resistant promoter and the regulatory gene arsR were obtained by PCR from the genome ofEscherichia coli DH5ct, andphiYFP was introduced into E. coli DH5ct as a reporter gene to construct an arsenic-resistant whole-cell biosensor （WCB-11） in which phiYFP was expressed well for the first time. Experimental results demonstrated that the biosensor has a good response to arsenic and the expression ofphiYFP. When strain WCB-11 was exposed to As^3＋ and As^5＋, the expression of yellow fluorescence was time-dependent and dose-dependent. This engineered construct is expected to become established as an inexpensive and convenient method for the detection of arsenic in the field.

关键词： arsenic reporter gene whole-cell biosensor yellow fluorescent protein (phiYFP)

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Improvement of a highly sensitive and specific whole-cell biosensor by adding a positive feedback amplifier

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Synthetic and Systems Biotechnology 2023年第2期8卷 292-299页

作者： Shuting Hu Guangbao Zhang Xiaoqiang Jia Department of Biochemical Engineering School of Chemical Engineering and TechnologyTianjin UniversityTianjin300072PR China Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering(Ministry of Education) School of Chemical Engineering and TechnologyTianjin UniversityTianjin300350PR China Collaborative Innovation Center of Chemical Science and Engineering(Tianjin) Tianjin300072PR China

In this study,we designed a Cd^(2+)whole-cell biosensor with both positive and negative feedback cascade am-plifiers in Pseudomonas putida KT2440(LTCM)based on our previous design with only a negative feedback amplifier(TCM).The results showed that the newly developed biosensor LTCM was greatly improved compared to ***,the linear response range of LTCM was expanded while the maximum linear response range was raised from 0.05 to 0.1μ***,adding a positive feedback amplifier further increased the fluorescence output signal of LTCM 1.11–2.64 times under the same culture ***,the response time of LTCM for detection of practical samples was reduced from 6 to 4 *** the same time,LTCM still retained very high sensitivity and specificity,while its lowest detection limit was 0.1 nM Cd^(2+)and the specificity was 23.29(compared to 0.1 nM and 17.55 in TCM,respectively).In summary,the positive and negative feedback cascade amplifiers effectively improved the performance of the biosensor LTCM,resulting in a greater linear response range,higher output signal intensity,and shorter response time than TCM while retaining comparable sensitivity and specificity,indicating better potential for practical applications.

关键词： Cadmium detection Positive feedback amplifier Sensitivity Specificity whole-cell biosensor

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Development of a whole-cell biosensor based on an ArsR-Pars regulatory circuit from Geobacter sulfurreducens

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Environmental Science and Ecotechnology 2021年第2期6卷 3-10页

作者： Pengsong Li Yumingzi Wang Xin Yuan Xinying Liu Chunmao Liu Xiaofen Fu Dezhi Sun Yan Dang Dawn E.Holmes Beijing Key Laboratory for Source Control Technology of Water Pollution Engineering Research Center for Water Pollution Source Control and EcoremediationCollege of Environmental Science and EngineeringBeijing Forestry UniversityBeijing 100083China Key Laboratory of Beijing Yanjing Beer Brewing Technology Beijing Yan Jing Brewery Company Ltd.Beijing 101300China MOST-USDA Joint Research Center for Biofuels Beijing Engineering Research Center for BiofuelsInstitute of New Energy TechnologyTsinghua UniversityBeijing 100084China Department of Physical and Biological Sciences Western New England University1215 Wilbraham RdSpringfieldMA 01119USA

In this study,an Escherichia coli(***)whole-cell biosensor for the specific detection of bioavailable arsenic was developed by placing a green fluorescent protein(GFP)reporter gene under the control of the ArsR1(GSU2952)regulatory circuit from Geobacter *** cells only emitted green fluorescence in the presence of arsenite and were more sensitive to arsenite when they were grown in M9 supplemented medium compared to LB *** optimal test conditions,the Geobacter arsR1 promoter had a detection limit of 0.01 mM arsenite and the GFP expression was linear within a range of 0.03-0.1 mM(2.25-7.5 mg/l).These values were well below World Health Organization’s drinking water quality standard,which is 10 mg/*** feasibility of using this whole-cell biosensor to detect arsenic in water samples,such as arsenic polluted tap water and landfill leachate was *** biosensor was determined to be just as sensitive as atomic fluorescence *** study examines the potential applications of biosensors constructed with Geobacter ArsR-Pars regulatory circuits and provides a rapid and cost-effective tool that can be used for arsenic detection in water samples.

关键词： Arsenic detection whole-cell biosensor Geobacter sulfurreducens ars operon

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Toxicity assessment of heavy metals and organic compounds using cellSense biosensor with ***

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Chinese Chemical Letters 2008年第2期19卷 211-214页

作者： Hong Wang Xue Jiang Wang Jian Fu Zhao Ling Chen State Key Laboratory of Pollution Control and Resource Reuse School of Environmental Science and Engineering ongji University Shanghai 200092 China

A new strategy using an arnperometric biosensor with Escherichia coli （E. coli） that provides a rapid toxicity determination of chemical compounds is described. The cellSense biosensor system comprises a biological component immobilized in intimate contact with a transducer which converts the biochemical signal into a quantifiable electrical signal. Toxicity assessment of heavy metals using *** biosensors could be finished within 30 min and the 50% effective concentrations （ECso） values of four heavy metals were determined. The results shows that inhibitory effects of four heavy metals to *** can be ranked in a decreasing order of Hg^2＋〉 Cu^2＋〉 Zn^2＋〉 Ni^2＋, which accords to the results of conventional bacterial counting method. The toxicity test of organic compounds by using cellSense biosensor was also demonstrated. The cellSense biosensor with E. coli shows a good, reproducible behavior and can be used for reproducible measurements.

关键词： whole-cell biosensor Heavy metal Organic compounds Toxicity

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