Construction of the Biosensor to Detect Arsenic in Water by Applying Technology Developed by PUCE-SI
Edith Verónica Mejía Segovia; Santiago Xavier Mafla Andrade; Diego Javier Jauregui Sierra; Moraima Cristina Mera Aguas
Abstract
Arsenic is a semimetal can form various organic and inorganic compounds, but its inorganic form is very toxic. At present there are several methods for the quantification of arsenic in water, such as: Atomic Absorption Spectroscopy, commercial Kits, among others which have a high power of resolution, sensitivity and reproducibility, however it can be expensive and difficult to operate. Bacterial biosensors emerge as an economically more accessible alternative, with potentially rapid measurements. The objective of this study was the development of a bacterial biosensor for the detection of arsenic using the modified Escherichia coli bacterium. This transformed strain has inserted plasmid pTOP Blunt V2, which confers resistance to ampicillin, for the realization of the plasmid the luminescence and arsenic resistance genes were synthesized. The Electroporation technique was used for the transformation and recombination of cells, which were subjected to controlled electrical impulses. Subsequently, it was cultured on LB agar and subjected to arsenic tolerance tests at concentrations of 00.010.020.04 and 0.08ppm as the sole source of energy. The absorbance of arsenic was determined by EPOCH microplate spectrophotometer methodology and Atomic Absorption Spectroscopy (EAA), resulting in the equation: y = 0.0005x + 0.1526 of the curve with R = 0.9975 with different absorbances and concentrations. Finally, it was possible to have a competent biosensor to measure of As concentrations from 0.01 to 0.08ppm with an error of 2.8% with respect to EAA measurement.
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References
BioTek (2017). BioTek. Obtenido de Microplate Data CollectionAnalysis Software: https://www.biotek.es/es/products/liquid-handling/
Dwivedi, S. (2016). Microorganism – As Biosensor for Arsenic detection. International Journal of Research in Science and Technology, 1-13.
Invitrogen (2014). En Invitrogen, Neon Transfection System.Thermo Fisher Scientic Inc. p. 4-15.
Lobos, C. P. (Noviembre de 2007). Uso de un Biosensor bacteriano, para detección de arsénico (en muestras ambientales).
MACROGEN Inc. (2018). Humanizing Genomics macrogen. Obtenido de Gene Synthesis Report: https://dna.macrogen.com/eng/quotation/ces/ces_quotaion_step1.jsp
Metalyser (2013). Metalyser-Catalogo-Trace-2o. Obtenido de http://ansam.mx/wp-content/ uploads/2014/04/Metalyser-Catalogo-Trace-2º.pdf
Microlab Industrial (2015). Absorcin atmica horno de grato. Obtenido de http://www.microlabindustrial.com/laboratorio/ver/metales/3/absorcion-atomica-horno-de-grato
Miller, J.; Miller, J. (1993). Estadistica para la Quimica Analitica. Capítulo 5 Errores en anlisis instrumental: Regresion y Correlacion. Addison-Wesley Iberoamericana. México, p. 87 -119.
Miller, J.; Miller, J. (2002). Estadistica y Quimiometria para Quimica Analitica. Capitulo 5 Metodos de Calibracion en Analisis Instrumental: Regresion y Correlacion, p. 11-152. Madrid, España: Prentice Hall.
Pingarrón, J. M. (2013). Biosensores amperométricos compósitos basados en peroxidasa. Available at, http://biblioteca.ucm.es/tesis/qui/ucm-t26582.pdf
Trang, P.; Berg, M.; Viet, P. H.; Mui, N. V. ,Meer, J. R. (2005). Bacterial bioassay for rapid and accurate analysis of arsenic in highly variable groundwater samples. Environmental science & technology 39(19), p. 25-30.
Valenuela, O. (2010). Evaluación de la concentración de Arsénico en agua empleando biosensores microbianos. Available at, http://tesis.bnct.ipn.mx:8080/ jspui/bitstream/123456789/9216/1/121.pdf
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