Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)
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:...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Veterinary World
2017-12-01
|
| Series: | Veterinary World |
| Subjects: | |
| Online Access: | http://www.veterinaryworld.org/Vol.10/December-2017/13.pdf |
| _version_ | 1818655979768643584 |
|---|---|
| author | Sakil Ahmed Aamir Sohail Sabiha Khatoon Shabnam Khan Mohammad Khalid Saifullah |
| author_facet | Sakil Ahmed Aamir Sohail Sabiha Khatoon Shabnam Khan Mohammad Khalid Saifullah |
| author_sort | Sakil Ahmed |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T03:18:18Z |
| format | Article |
| id | doaj.art-9123063a6f6a45d4849f6a19cfbbb02d |
| institution | Directory Open Access Journal |
| issn | 0972-8988 2231-0916 |
| language | English |
| last_indexed | 2024-12-17T03:18:18Z |
| publishDate | 2017-12-01 |
| publisher | Veterinary World |
| record_format | Article |
| series | Veterinary World |
| spelling | doaj.art-9123063a6f6a45d4849f6a19cfbbb02d2022-12-21T22:05:35ZengVeterinary WorldVeterinary World0972-89882231-09162017-12-0110121493150010.14202/vetworld.2017.1493-1500Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)Sakil Ahmed0Aamir Sohail1Sabiha Khatoon2Shabnam Khan3Mohammad Khalid Saifullah4Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.Department of Biochemistry, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.Section of Parasitology, Department of Zoology, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.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.http://www.veterinaryworld.org/Vol.10/December-2017/13.pdfBubalus bubalisGastrothylax crumeniferglutathione S-transferasepurificationsomatic tissue |
| spellingShingle | Sakil Ahmed Aamir Sohail Sabiha Khatoon Shabnam Khan Mohammad Khalid Saifullah Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea) Veterinary World Bubalus bubalis Gastrothylax crumenifer glutathione S-transferase purification somatic tissue |
| title | Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea) |
| title_full | Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea) |
| title_fullStr | Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea) |
| title_full_unstemmed | Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea) |
| title_short | Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea) |
| title_sort | partial purification and characterization of glutathione s transferase from the somatic tissue of gastrothylax crumenifer trematoda digenea |
| topic | Bubalus bubalis Gastrothylax crumenifer glutathione S-transferase purification somatic tissue |
| url | http://www.veterinaryworld.org/Vol.10/December-2017/13.pdf |
| work_keys_str_mv | AT sakilahmed partialpurificationandcharacterizationofglutathionestransferasefromthesomatictissueofgastrothylaxcrumenifertrematodadigenea AT aamirsohail partialpurificationandcharacterizationofglutathionestransferasefromthesomatictissueofgastrothylaxcrumenifertrematodadigenea AT sabihakhatoon partialpurificationandcharacterizationofglutathionestransferasefromthesomatictissueofgastrothylaxcrumenifertrematodadigenea AT shabnamkhan partialpurificationandcharacterizationofglutathionestransferasefromthesomatictissueofgastrothylaxcrumenifertrematodadigenea AT mohammadkhalidsaifullah partialpurificationandcharacterizationofglutathionestransferasefromthesomatictissueofgastrothylaxcrumenifertrematodadigenea |