PRELIMINARY INVESTIGATIONS ON AN ENZYME IMMOBILIZED OPTICAL BIOSENSOR FOR ARSENITE DETECTION

Authors

  • Wei Kheng Teoh Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia
  • Farhana Zahari Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia
  • Shafinaz Shahir Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia

DOI:

https://doi.org/10.11113/jt.v78.7757

Keywords:

Arsenite, biosensor, arsenite oxidase, recombinant, DCPIP

Abstract

Arsenite is an inorganic form of arsenic that poses hazardous effect to human. It is a common environmental heavy metal contaminant ubiquitously found in water and groundwater. In this study, an optical biosensor for arsenite determination was developed by immobilization of crude arsenite oxidase (Aio) extracted from recombinant E. coli, in chitosan solution coated on triacetyl-cellulose membrane employing DCPIP as colour indicator. The arsenite oxidase (Aio) was successfully expressed and extracted from recombinant E. coli strain BL21 (DE3). The protein concentration and specific activity of the crude arsenite oxidase were determined.  Expression of Aio was confirmed by SDS-PAGE. The crude Aio was also successfully immobilized in chitosan and coated on triacetyl cellulose membrane. The response time and dynamic range of the optical biosensor were optimized. The response time of the developed biosensor was 15 minutes. The amount of DCPIP reduced (DA) was inversely proportional to the arsenite concentration. Standard calibration curve for arsenite detection was achieved within the range of arsenite concentration from 25 µM to 200 µM. The maximum detection limit was determined to be 250 µM arsenite.

Author Biographies

  • Wei Kheng Teoh, Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia
    Dept of Biosciences and Health Sciences
  • Farhana Zahari, Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia
    Dept of Biosciences and Health Sciences
  • Shafinaz Shahir, Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Johor, Malaysia
    Dept of Biosciences and Health SciencesDept of Biosciences and Health Sciences

References

Abdullah, J., Ahmad, M., Karuppiah, N., Heng, L. Y. and Sidek, H. 2006. Immobilization Of Tyrosinase In Chitosan Film For An Optical Detection Of Phenol. Sensors and Actuators B: Chemical. 114(2): 604-609.

Anderson, G. L., Williams, J. and Hille, R. 1992. The Purification And Characterization Of Arsenite Oxidase From Alcaligenes Faecalis, A Molybdenum-Containing Hydroxylase. Journal of Biological Chemistry. 267(33): 23674-82.

Battaglia-Brunet, F., Joulian, C., Garrido, F., Dictor, M.-C., Morin, D., Coupland, K., Barrie Johnson, D., Hallberg, K. and Baranger, P. 2006. Oxidation Of Arsenite By Thiomonas Strains And Characterization of Thiomonas arsenivorans sp. nov. Antonie van Leeuwenhoek. 89(1): 99-108.

Ensafi, A. A. and Amini, M. 2012. Highly Selective Optical Nitrite Sensor For Food Analysis Based On Lauth’s Violet–Triacetyl Cellulose Membrane Film. Food Chemistry. 132(3): 1600-1606.

Hu, Q., Li, L., Wang, Y., Zhao, W., Qi, H. and Zhuang, G. 2010. Construction of WCB-11: A Novel Phiyfp Arsenic-Resistant Whole-Cell Biosensor. Journal of Environmental Sciences. 22(9): 1469-1474.

Majumder, A., Bhattacharyya, K., Bhattacharyya, S. and Kole, S. C. 2013. Arsenic-Tolerant, Arsenite-Oxidising Bacterial Strains In The Contaminated Soils Of West Bengal, India. Science of the Total Environment. 463-464: 1006-1014.

Male, K. B., Hrapovic, S., Santini, J. M. and Luong, J. H. T. 2007. Biosensor for Arsenite Using Arsenite Oxidase and Multiwalled Carbon Nanotube Modified Electrodes. Anal. Chem. 79: 7831-7837.

Ng, J. C., Wang, J. and Shraim, A. 2003. A Global Health Problem Caused By Arsenic From Natural Sources. Chemosphere. 52(9): 1353-1359.

Nguyen, V. A., Bang, S., Viet, P. H. and Kim, K.-W. 2009. Contamination Of Groundwater And Risk Assessment For Arsenic Exposure In Ha Nam Province, Vietnam. Environment International. 35(3): 466-472.

Rasul, S. B., Hossain, Z., Munir, A. K. M., Alauddin, M., Khan, A. H. and Hussam, A. 2002. Electrochemical Measurement And Speciation Of Inorganic Arsenic In Groundwater Of Bangladesh. Talanta. 58(1): 33-43.

Roberto, F. F., Barnes, J. M. and Bruhn, D. F. 2002. Evaluation Of A GFP Reporter Gene Construct For Environmental Arsenic Detection. Talanta. 58(1): 181-188.

Routh, J. and Hjelmquist, P. 2011. Distribution Of Arsenic And Its Mobility In Shallow Aquifer Sediments From Ambikanagar, West Bengal, India. Applied Geochemistry. 26(4): 505-515.

Shafinaz Shahir, Farhana Zahari and Wei Kheng Teoh. 2016. Enzyme Based Biosensor For Arsenite Detection. In Wan Rosmiza Wan Dagang and Fazilah Abd Manan (ed.). Sustainable Biological Approaches for Wastes Remediation. Malaysia: Penerbit UTM Press. 37-46.

Silver, S. and Phung, L. T. 2005. Genes and Enzymes Involved in Bacterial Oxidation and Reduction of Inorganic Arsenic. Appl. Environ. Microbiol. 71(2): 599-608.

Thévenot, D. R., Toth, K., Durst, R. A. and Wilson, G. S. 2001. Electrochemical Biosensors: Recommended Definitions And Classification. Biosensors and Bioelectronics. 16(1-2): 121-131.

Yi, H., Wu, L.-Q., Sumner, J. J., Gillespie, J. B., Payne, G. F. and Bentley, W. E. 2003. Chitosan Scaffolds For Biomolecular Assembly: Coupling Nucleic Acid Probes For Detecting Hybridization. Biotechnology and Bioengineering. 83(6): 646-652.

Downloads

Published

2016-09-29

Issue

Section

Science and Engineering

How to Cite

PRELIMINARY INVESTIGATIONS ON AN ENZYME IMMOBILIZED OPTICAL BIOSENSOR FOR ARSENITE DETECTION. (2016). Jurnal Teknologi, 78(10). https://doi.org/10.11113/jt.v78.7757