Partial Purification of Cholinesterase from Pangasius pangasius using Affinity Chromatography

Authors

  • Lik Gin Tham Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia.
  • Mohd Yunus Shukor Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia.
  • Mohd Arif Syed Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia.
  • Nor Aripin Shamaan Faculty of Medicine and Health Sciences, Universiti Sains Islam Malaysia, 13th Floor, Menara B, Persiaran MPAJ, Jalan Pandan Utama, Pandan Indah, 55100 Kuala Lumpur, Malaysia.
  • Ahmad Razi Othman Research Centre for Sustainable Process Technology (CESPRO), Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

DOI:

https://doi.org/10.54987/jemat.v5i2.410

Keywords:

Pangasius pangasius, partial purification, affinity chromatography, procainamide, acetylcholinesterase

Abstract

An affinity chromatography column to purify AChE was developed using procainamide as the ligand coupled with the epoxy activated Sephacryl-S1000 as the modified matrix. From this study, the procainamide matrix was found to be capable of binding acetylcholinesterase (AChE) from the brain of Pangasius pangasius efficiently, with percentage of yield of approximately 51.97% and a purification fold of 7.52. The value for Michaelis-Menten (Km app) was 0.1265 mM and maximal velocity (Vmax app) of the partially purified enzyme was 0.6981 µmol/min/mg, with a substrate specificity in the sequence of— acetylthiocholine iodide (ATC) > propinylcholine iodide (PTC) > butyrylcholine iodide (BTC). This strongly suggest that using this column, AChE can be purified successfully and efficiently. SDS-PAGE indicated a reduced band intensity, indicating that the procainamide-based affinity chromatography column produces acceptable partially purified enzyme adequate for the application in the rapid detection of carbamate and organophosphate (OP) at the large scale.

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Published

31.12.2017

How to Cite

Tham, L. G., Shukor, M. Y., Syed, M. A., Shamaan, N. A., & Othman, A. R. (2017). Partial Purification of Cholinesterase from Pangasius pangasius using Affinity Chromatography. Journal of Environmental Microbiology and Toxicology, 5(2), 14–18. https://doi.org/10.54987/jemat.v5i2.410

Issue

Vol. 5 No. 2 (2017)

Section

Articles

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