* Corresponding Author
Kinetic Evaluation of the Inhibition of Acetylcholinesterase for use as a biosensor
https://doi.org/10.22146/ajche.56709 R. Bhuvanagayathri(1), David K Daniel(2*), Gnanasundaram Nirmala(3)
(1) School of Chemical Engineering, Vellore Institute of Technology, Vellore-632014, Tamil Nadu, India.
(2) Department of Chemical Engineering, Amal Jyothi College of Engineering, Kanjirappally, Kottayam-686518 Kerala, India.
(3) School of Chemical Engineering, Vellore Institute of Technology, Vellore-632014, Tamil Nadu, India.
(*) Corresponding Author
Abstract
The release of pesticides into the environment has increased, and there is a lack of monitoring of these contaminants. Since the conventional methods of monitoring these contaminants are complicated, costly and time-consuming, mechanisms based on acetylcholinesterase inhibition have emerged as simple and rapid tools for such applications. However, theacetylcholinesterase’s effectiveness as a sensing element in such biosensor systems depends on the conditions selected to measure acetylcholinesterase activity and the concentration of substrate or inhibitor, which in turn affect the reaction rates. Therefore, in the present work, the factors affecting the acetylcholinesterase activity were investigated and inhibition experiments were carried out to evaluate the kinetic parameters. The inhibition rate constant for acetylcholinesterase Ki was found to be 1.9 ppm. The kinetic parameter Km was found to be 3.8mM and Vmax was found to be 1.3µM/min from the Eadie-Hofstee plot. The kinetic study using Lineweaver-Burk method showed mixed type of inhibition of acetylcholinesterase with carbofuran.
Keywords
Acetylcholinesterase; Biosensor; Inhibition; Kinetics
Full Text:
PDFReferences
- Abhilash, P. C., and Nandita S., (2009), “Pesticide use and application: An Indian scenario,” J. Hazard Mater., 165, 1-12.
- Bhuvanagayathri, R., David, K. D., Albert, I. K., (2018). “Progress in enzyme inhibition based detection of pesticides, Engineering in Life Sciences,” 18, 4-19.
- Bruce, C. L., James, C. M., Robert, C. H., and Glenn, H. F., (1983). “Determination of carbosulfan and carbofuran residues in plants, soil, and water by gas chromatography,” J. Agric. Food Chem., 31, 220-223.
- Cometa, M. F., Lorenzini, P., Fortuna, S., Volpe, M. T., Meneguz, A., and Palmery, M., (2005). “Invitro inhibitory effect of aflatoxin B1on acetylcholinesterase activity in mouse brain,” Toxicology, 206(1),125-35
- Ellman, G. L., Courtney, D. K., Andres, V., and Featherstone, R. M., (1961). “A new and rapid colorimetric determination of acetylchoinesterase activity,” Biochem. Pharmacol., 7, 88-95.
- Fatma, N. K., Faruk, B., and Vasif H., (2002). “Construction of an acetylcholinesterase –choline oxidase biosensor for aldicarb determination,” Biosens Bioelectron, 17, 531- 539.
- Ferat, S., Gokhan, D., Hayrettin, T., (2005). “A novel matrix for the immobilization of acetylcholinesterase,” Int. J. Biol. Macromol., 37, 148-153.
- Fuad, A., (2016). “A HPLC-UV method for determination of three pesticides in water,” Int. J. of Advances in Chemistry, 2 (1), 9-16.
- Ghorab, M. A., and Khalil, M. S., (2016). “The effect of pesticides pollution on our Life and environment,” J. of Pollution Effects and Control, 4, 159.
- Gilvanda, S. N., Petrskladal, Hideko, Y., Damia, B., (1998). “Determination of carbamate residues in crop samples by cholinesterase based biosensors and chromatographic techniques,” Anal. Chim. Acta, 362, 59-68.
- Jose, F. H., Ana, M., (2008). “Determination of N-methyl carbamate pesticides in water and vegetable samples by HPLC with post-column chemiluminescence detection using the luminol reaction,” Anal. Chim. Acta., 630, 194-204.
- Lanny, S., Olivia, P.D., and Liliana, L., (2017). “Kinetics of oil-in-water emulsion stabilization using lecithin and biosilica,” Asean Journal of Chemical Engineering, 17(1),1-7.
- 13. Lineweaver, H., and Burk, D. (1934). "The determination of enzyme dissociation constants". J. Amer. Chem. Soc., 56 (3), 658–666.
- Marinov, I., Gabrovska, K., Velichkova, J., and Godjevargova, T., (2009). “Immobilization of acetylcholinesterase on nanostructure polyacrylonitrile membranes,” Int. J. of Biological Macromolecules, 44(4), 338-345.
- Roy, F. T., (1990). “Mechanism of Action of organophosphorus and carbamates insecticides,” Environ. Health Persp., 87, 245-24.
- Seyed V. S., (2018). “Ellman’s Method is still an appropriate method for measurement of cholinesterases activities,” EXCLI Journal 2018, 17, 798-799.
- Serge, B., (1985). “Determination of acetylcholinesterase activity by a new chemiluminescence assay with natural substrate,” Biochem. J., 225, 825-828.
- Shuaping, Z., Lian-Gang, S., Yi – Zhing, Lli – Yishi., (2008). “Study of enzyme biosensor for monitoring carbamate pesticides in sea water,” IFMBE Proceedings, 19, 323 -325.
- Siriwuan, S., Proespichaya, K., Punnee, A., Chusak L., Booncharoen, W., Panote T., (2005). “Semidisposable reactor biosensor for detecting carbamate pesticides in water,” Biosens. Bioelectron., 21, 445-454.
- Songci, X., Aibo, W., Haode, C., and Yang, Z., (2007). “Production of a novel recombinant Drosophila melanogaster acetyl-cholinesterase for detection of organophosphosphate and carbamate insecticide residues,” Biomolecular Engineering, 24 (2), 253-261.
- Vangelis, G. A., Yannis, D. C., (2002). “A portable fibre-optic pesticides biosensor based on immobilized cholinesterase and sol-gel entrapped bromocresol purple for in-field use,” Biosens. Bioelectron. 17, 61-69.
- Valber, A. P., Josiane, C., Sergio, A.S. M., Mauro, B., (2008). “Determination of Parathion and Carbaryl pesticides in water and food samples using a self assembled monolayer/Acetylcholinesterase Electro-chemical Biosensor,” Sensors, 8, 4600 -4610.
- Zhi, X., Zhibo, H., Jinhui, L., and Defang, X., (2017). "Simultaneous Determination of Eight Pesticide Residues in Cowpeas by GC–ECD," J. of Chromatographic Science, 55 (1), 1–6.
DOI: https://doi.org/10.22146/ajche.56709
Article Metrics
Abstract views : 4871
| 
views : 7499
Refbacks
- There are currently no refbacks.