alexa Purification and Characterization of Glutathione-S-Transferase from Rats Liver: Effect of Carbon Tetrachloride and Camels Milk | OMICS International | Abstract
ISSN: 2157-7064

Journal of Chromatography & Separation Techniques
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Research Article

Purification and Characterization of Glutathione-S-Transferase from Rats Liver: Effect of Carbon Tetrachloride and Camels Milk

Mohamed Lebda1*, Nabil Taha1, Saad Noeman3, Mahdy Korshom1, Abd El-Wahab Mandour1 and Ahlam El-Leboudy2

1Department of Biochemistry, Faculty of Veterinary Medicine, Alexandria University, Egypt

2Department of Food Hygiene, Faculty of Veterinary Medicine, Alexandria University, Egypt

3Department of Medical Biochemistry, Faculty of Medicine, Tanta University, Egypt

*Corresponding Author:
Mohamed Lebda
Department of Biochemistry
Faculty of Veterinary Medicine
Alexandria University, Egypt
Tel: +20 10 08479197
Fax: +20 45 2960450
E-mail: [email protected]

Received date: July 11, 2012; Accepted date: August 16, 2012; Published date: August 18, 2012

Citation: Lebda M, Taha N, Noeman S, Korshom M, El-Wahab Mandour A, et al.(2012) Purification and Characterization of Glutathione-S-Transferase from Rat’s Liver: Effect of Carbon Tetrachloride and Camel’s Milk. J Chromat Separation Techniq 3:133.. doi:10.4172/2157-7064.1000133

Copyright: © 2012 Lebda M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited


Glutathione-S-transferases (GSTs) constitute a multifunctional family of dimeric and mainly cytosolic biotransformation enzymes which play an important role in protecting tissues from oxidative stress. They catalyzed the conjugation of intracellular glutathione with great varieties of chemicals possessing electrophilic centre and the final GSH-conjugates had increased hydrophilicity which facilitate their further metabolism and elimination. Our study was carried out to purify GST enzyme from rat’s liver using one step affinity chromatography and investigate the inhibitory effect of carbon tetrachloride and ameliorating effect of camel’s milk administration. The purification of GST enzyme revealed that the protein concentration decreased in GSH affinity filtrate than the crude extract while the specific enzyme activity increased indicating that the single purification step cut off the other proteins and conserve the protein of GST enzyme. CCL4 decrease the Vmax and increase Km for both GSH and CDNB substrates than control indicating that CCL4 may be uncompetitive inhibitor of GST enzyme while camel’s milk increase the Vmax and decrease Km for both GSH and CDNB substrates indicating that camel’s milk increase the affinity of enzyme to both substrates. The activity of GST increased by increasing the substrate concentration till reaching the maximum activity after which the activity of GST forming a plateau. Regarding the effect of temperature on GST activity, the maximum GST activity was observed between 25-35ºC and inactivated over 40ºC. The GST activity increased when pH increased progressively till reach the maximum activity at pH value 7.1 and decreased when the pH increased. Our conclusion indicating that CCL4 is uncompetitive inhibitor of GST enzyme while camel’s milk increases the activity and affinity of enzyme toward its both substrates.


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