Development of a glutathione-S-transferase-based biosensor for the detection of heavy metals

Saatçi, Ebru
In the recent years, environmental pollution becomes a health threatening issue for human beings. Technological developments introduce industrial wastes and heavy metals, and developments in agriculture introduce pesticides into the world that we live. All these toxic wastes accumulate in drinking water and food consumed by humans. Therefore, detection of toxic wastes in all kinds of environmental samples, and development of new detection techniques become an important issue. In this study, development of a protein-based biosensor for detection of heavy metals in environmental samples, by expressing genetically modified glutathione S-transferase (GST-(His)6) protein in E.Coli BL21 (DE3) expression system, was designed. Recombinant GST proteins was expressed in E.Coli BL21 (DE3) expression system and purified with Glutathione Sepharose 4B affinity column and Ni-NTA spin kit. GST activities were determined using the GST substrate 1-chloro-2,4-dinitrobenzene (CDNB). Protein expression was tested by SDS-PAGE and Western blot analysis. Product formation linearly increased up to 1 mM CDNB, 1 mM GSH, 1.7 æg proteins in 0.05 M, pH 6.9 phosphate buffer in the final volume of 1.0 ml at 25?C. The Vmax and Km values for GST-(His)6 towards CDNB and GSH were calculated with Lineweaver-Burk as CDNB Vmax; 22.88 æmol/min/mg, Km; 4.29 mM,and as GSH Vmax; 6.42 æmol/min/mg, 24.45 æmol/min/mg, Km; 3.69 mM, respectively. Biosensor working electrode was prepared by immobilizing the GST-(His)6 by 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDC) coupling on the gold surface. Electrode preparation was confirmed by cyclic voltammetry measurements. The biosensor was inserted as the working electrode in the constructed three(four)-electrode flow cell. The conformational change resulting from the binding of the metal ions to the recombinant protein causing a capacitance change


Identification and Elucidation of the Designing and Operational Issues of Trickling Filter Systems for Wastewater Treatment
Ali, Imran; Khan, Zahid M.; Peng, Changsheng; Naz, Iffat; Sultan, Muhammad; Ali, Mohsin; Mahmood, Muhammad H.; Niaz, Yasir (2017-01-01)
Water pollution has become a major environmental concern for public and environmental health in developing countries. Water resources are being contaminated mainly due to mixing of domestic, municipal, and industrial wastewaters. The wastewater management and treatment situation is deplorable mainly because of financial constraints, the unavailability of technically trained human resources, and electricity shortages. Moreover, there is a challenge for the scientific community and wastewater management exper...
Computer modeling of blowback oil consumption in internal engines
Bilge, Egemen; Dursunkaya, Zafer; Department of Mechanical Engineering (2009)
Environmental pollution is an important problem of our world. Governments are aware of this problem and emission regulations are continuously improved. One of the strictest regulations is about unburned and burned hydrocarbon emissions. In internal combustion engines the origin of the burned and unburned hydrocarbons is fuel and engine oil. As a result of the sanctions and the necessity of improved combustion performance of the engine, manufacturers work on manufacturing technology and engine tribology. Wit...
Formation of nitrogen functionalities in biochar materials and their role in the mitigation of hazardous emerging organic pollutants from wastewater
Başer, Begüm; Yousaf, Balal; Yetiş, Ülkü; Abbas, Qumber; Kwon, Eilhann E.; Wang, Shengsen; Bolan, Nanthi S.; Rinklebe, Jörg (2021-08-15)
Emerging organic pollutants (EOPs) are serious environmental concerns known for their prominent adverse and hazardous ecological effects, and persistence in nature. Their detrimental impacts have inspired researchers to develop the strategic tools that reduce and overcome the challenges caused by EOPs' rising concentration. As such, biochar becomes as a promising class of biomass-derived functional materials that can be used as low-cost and environmentally-friendly emerging catalysts to remove EOPs. Herein,...
Development of Simulation-Optimization Model (MUSIC-GA) for Urban Stormwater Management
Montaseri, Majid; Afshar, Mahdı Hesamı ; Bozorg-Haddad, Omid (2015-10-01)
Urban development leads to replacement of impervious surfaces with pervious surfaces, vegetation and production of various environmental pollutants. In addition, this phenomenon has increased the devastating effects of urban stormwater and has exacerbated the water quality of many freshwater resources (surface and underground) throughout the world. To alleviate this condition, new models of urban stormwater systems, including Water Sensitive Urban Design (WSUD) as well as its software package, MUSIC, have b...
Evaluation of Performance of Arsenic Bioreporter Immobilized Electrospun Membranes for Arsenic Detection in Water
Arık Kınalı, Nehir; Öktem, Hüseyin Avni; Tezcaner, Ayşen; Department of Molecular Biology and Genetics (2022-12-9)
Heavy metal pollution, which is growing and threatening the health of all living things as well as human health, can lead to serious lifelong consequences. It is very important to detect and monitor these heavy metals that cause environmental pollution. In addition to traditional laboratory techniques, the use of bacterial biosensors to detect heavy metals has come into focus. In addition, it is known that the immobilization of these bacterial biosensors is more advantageous in many respects, such as being ...
Citation Formats
E. Saatçi, “Development of a glutathione-S-transferase-based biosensor for the detection of heavy metals,” Ph.D. - Doctoral Program, Middle East Technical University, 2005.