Open Access
Research (Published online: 21-12-2017)
13. Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)
Sakil Ahmed, Aamir Sohail, Sabiha Khatoon, Shabnam Khan and Mohammad Khalid Saifullah
Veterinary World, 10(12): 1493-1500

Sakil Ahmed: Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Aamir Sohail: Department of Biochemistry, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Sabiha Khatoon: Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Shabnam Khan: Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.
Mohammad Khalid Saifullah: Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.

doi: 10.14202/vetworld.2017.1493-1500

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

Corresponding author: Mohammad Khalid Saifullah


Citation: 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.

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.


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