Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)

Vet World Vol.10 December-2017 Article-13

Research Article

Veterinary World, 10(12): 1493-1500

https://doi.org/10.14202/vetworld.2017.1493-1500

Sakil Ahmed¹, Aamir Sohail², Sabiha Khatoon¹, Shabnam Khan¹, and Mohammad Khalid Saifullah¹

1. Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
2. Department of Biochemistry, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.

Background and Aim: Aim of the present study was to carry out the partial purification and biochemical characterization of glutathione S-transferase (GST) from the somatic tissue of ruminal amphistome parasite, Gastrothylax crumenifer (Gc) infecting Indian water buffalo (Bubalus bubalis).

Materials and Methods: The crude somatic homogenate of Gc was subjected to progressive ammonium sulfate precipitation followed by size exclusion chromatography in a Sephacryl S 100-HR column. The partially purified GST was assayed spectrophotometrically, and the corresponding enzyme activity was also recorded in polyacrylamide gel. GST isolated from the amphistome parasite was also exposed to variable changes in temperature and the pH gradient of the assay mixture.

Results: The precipitated amphistome GST molecules showed maximum activity in the sixth elution fraction. The GST subunit appeared as a single band in the reducing polyacrylamide gel electrophoresis with an apparent molecular weight of 26 kDa. The GST proteins were found to be fairly stable up to 37°C, beyond this the activity got heavily impaired. Further, the GST obtained showed a pH optima of 7.5.

Conclusion: Present findings showed that GST from Gc could be conveniently purified using gel filtration chromatography. The purified enzyme showed maximum stability and activity at 4°C. Keywords: Bubalus bubalis, Gastrothylax crumenifer, glutathione S-transferase, purification, somatic tissue.

Keywords: Bubalus bubalis, Gastrothylax crumenifer, glutathione S-transferase, purification, somatic tissue.

How to cite this article: Ahmed S, Sohail A, Khatoon S, Khan S, Saifullah MK (2017) Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea), Veterinary World, 10(12): 1493-1500.

Received: 19-08-2017 Accepted: 15-11-2017 Published online: 21-12-2017

Corresponding author: Mohammad Khalid Saifullah E-mail: mkhalidsaif@yahoo.com

DOI: 10.14202/vetworld.2017.1493-1500

Copyright: Ahmed, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.