Characterisation of purified acetylcholinesterase (EC 3.1.1.7) from <i>Oreochromis mossambica</i> brain tissues

Authors

  • Lik Gin Tham Dept of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
  • Natarajan Perumal Dept of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
  • Siti Aqlima Ahmad Dept of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
  • Mohd Khalizan Sabullah Faculty of Science and Natural Resources, Universiti Malaysia Sabah, 88400 Kota Kinabalu, Sabah, Malaysia.

DOI:

https://doi.org/10.54987/jobimb.v5i2.346

Keywords:

acetylcholinesterase (AChE); purification; biochemical characterization; Oreochromis mossambica

Abstract

This study reports on the characterization of a purified AChE from Oreochromis mossambica brain extract. The purification protocol involved the application of custom-synthesized affinity chromatography gel (Edrophonium–Sephacryl S-400) and the use of high performance liquid chromatography system (HPLC). Soluble AChE was partially purified 27.9 fold with a highest specific activity around 73.1 × 103 U/mg proteins. The partially purified AChE higher capability to hydrolyse acetylthiocholine (ATC) and shows less degradation against propionylthiocholine (PTC) and also butyrylthiocholine (BTC). Based on enzyme kinetic analysis, the partially purified AChE exhibits the apparent Michaelis constants Km, for ATC, PTC and BTC in the range of 125, 260 and 600 µM and the maximum velocities Vmax were 276, 59 and 36 µmol/min/mg protein, respectively. The apparent inhibition constant (ki) values of eserine, propidium and carbofuran were 0.24 µM-1min-1, 65 µM-1min-1 and 0.41 µM-1min-1 µM-1min-1, respectively. The purified enzyme is apparently an AChE since it capable to hydrolyzes ATC at a higher rate compared to other synthetic substrates, at pH 7.0 and 25ºC, and is inhibited by it specific inhibitor which is eserine but not by iso-OMPA.

Downloads

Published

31.12.2017

How to Cite

Tham, L. G., Perumal, N., Ahmad, S. A., & Sabullah, M. K. (2017). Characterisation of purified acetylcholinesterase (EC 3.1.1.7) from <i>Oreochromis mossambica</i> brain tissues. Journal of Biochemistry, Microbiology and Biotechnology, 5(2), 22–27. https://doi.org/10.54987/jobimb.v5i2.346

Issue

Vol. 5 No. 2 (2017)

Section

Articles

License

Authors who publish with this journal agree to the following terms:

    1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
    2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
    3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).