Polyacrylamide hydrogel encapsulated E. coli expressing metal-sensing green fluorescent protein as a potential tool for copper ion determination

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dc.contributor.authorTantimongcolwat, Tanawut
dc.contributor.authorIsarankura-Na-Ayudhya, Chartchalerm
dc.contributor.authorSrisarin, Apapan
dc.contributor.authorGalla, Hans-Joachim
dc.contributor.authorPrachayasittikul, Virapong
dc.date.accessioned2014-07-24T09:03:13Z
dc.date.available2014-07-24T09:03:13Z
dc.date.issued2014-04-07
dc.description.abstractA simple, inexpensive and field applicable metal determination system would be a powerful tool for the efficient control of metal ion contamination in various sources e.g. drinking-water, water reservoir and waste discharges. In this study, we developed a cell-based metal sensor for specific and real-time detection of copper ions. E. coli expressing metal-sensing green fluorescent protein (designated as TG1/(CG)6GFP and TG1/H6CdBP4GFP) were constructed and served as a metal analytical system. Copper ions were found to exert a fluorescence quenching effect, while zinc and cadmium ions caused minor fluorescence enhancement in the engineered bacterial suspension. To construct a user-friendly and reagentless metal detection system, TG1/H6CdBP4GFP and TG1/(CG)6GFP were encapsulated in polyacrylamide hydrogels that were subsequently immobilized on an optical fiber equipped with a fluorescence detection module. The sensor could be applied to measure metal ions by simply dipping the encapsulated bacteria into a metal solution and monitoring fluorescence changes in real time as a function of the metal concentration in solution. The sensor system demonstrated high specificity toward copper ions. The fluorescence intensities of the encapsulated TG1/(CG)6GFP and TG1/H6CdBP4GFP were quenched by approximately 70 % and 80 % by a high-dose of copper ions (50mM), respectively. The level of fluorescence quenching exhibited a direct correlation with the copper concentration, with a linear correlation coefficient (r) of 0.99. The cell-based metal sensor was able to efficiently monitor copper concentrations ranging between 5 µM and 50 mM, encompassing the maximum allowed copper contamination in drinking water (31.15 µM) established by the WHO. Furthermore, the cell-based metal sensor could undergo prolonged storage for at least 2 weeks without significantly influencing the copper sensitivity.en
dc.identifier.issn1611-2156
dc.identifier.urihttp://hdl.handle.net/2003/33528
dc.identifier.urihttp://dx.doi.org/10.17877/DE290R-15513
dc.language.isoen
dc.relation.ispartofseriesEXCLI Journal ; Vol. 13, 2014en
dc.subjectgreen fluorescent proteinen
dc.subjectmetal sensoren
dc.subjectcopperen
dc.subjectoptical sensoren
dc.subject.ddc610
dc.titlePolyacrylamide hydrogel encapsulated E. coli expressing metal-sensing green fluorescent protein as a potential tool for copper ion determinationen
dc.typeText
dc.type.publicationtypearticle
dcterms.accessRightsopen access
eldorado.dnb.zdberstkatid2132560-1

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