Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver

Biotransformation and detoxification process in living organisms consists of two phases, phase I and phase 11. Phase I involves in the introduction of functional group into molecule while the phase I1 involves the conjugation of phase I metabolites. In phase 11, glutathione S-transferases (GSTs;...

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Main Author:	Saleh Huddin, Lailatul Jumaiyah
Format:	Thesis
Language:	English English
Published:	2006
Subjects:	Glutathione transferase - Kedah - Cattle Glutathione transferase - Kelantan - Cattle Glutathione transferase - Kedah - Water buffalo
Online Access:	http://psasir.upm.edu.my/id/eprint/5978/1/FBSB_2006_5.pdf
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spelling	my-upm-ir.59782023-01-03T06:52:33Z Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver 2006-04 Saleh Huddin, Lailatul Jumaiyah Biotransformation and detoxification process in living organisms consists of two phases, phase I and phase 11. Phase I involves in the introduction of functional group into molecule while the phase I1 involves the conjugation of phase I metabolites. In phase 11, glutathione S-transferases (GSTs; EC 2.5.1.18) has aroused much interest because of its involvement in the biotransformation and detoxification of wide spectrum of xenobiotics which can be from pesticides, herbicides and insecticides. The present study was undertaken to puriG and characterized cytosolic GSTs from livers of Kedah-Kelantan cattle (Bos indicus) and Malaysia water buffalo (Bubalus bubalis). The glutathione Stransferases were isolated from two important livestock livers, Kedah-Kelantan cattle (Bos indicus) and Malaysian water buffalo (Bubalus bubalis) by glutathione affinity chromatography. The affinity-glutathione chromatography successfully purifies the GSTs isoenzymes with 14.73% yield (62.77 purification fold) and 19.71 % yield (20.44 purification fold) for KK cattle and water buffalo livers respectively. Initial methods of purification included centrifugation and ultracentrifugation. The affinity elution with highest activity towards CDNB was estimated for the pI values using isoelectric focusing method via LKB-8 100 ampholyte type (LKB Bromna) apparatus. pI values for affinity purified KK cattle liver are 5.7 (C-34), 6.9 (C-38) and 8.8 (C-42). While for the water buffalo liver, the pI values for glutathione affinity purified isoenzymes are 6.85 (B-23) and 7.2 (B-24). The isoenzymes were then tested using SDS-PAGE method for purity and also to estimate the molecular weight estimation. It has been estimated that molecular weight for water buffalo isoenzymes of B-23 was 29.3 * 0.05 kDa and B-24 was 30.74 * 0.16 kDa. The KK cattle liver isoenzymes molecular weight was estimated with C-34 was 29.9 * 0.14; C-38 was 28.3 * 0.09 and 27.7 *0.03 for C-42. The study showed that KK cattle liver GSTs exist as isoenzymes (PI 8.8, 6.9 and 5.7), and have high activity towards CDNB, low towards DCNB and no activity towards the ethacrynic acid for the substrate specificities. On the other hand, the water buffalo liver GSTs exist as isoenzymes with pI 6.85 and 7.2. For the substrate specificities, the isoenzymes also have high activity for CDNB, but low for DCNB and could not be detected for the ethacrynic acid. Glutathione transferase - Kedah - Cattle Glutathione transferase - Kelantan - Cattle Glutathione transferase - Kedah - Water buffalo 2006-04 Thesis http://psasir.upm.edu.my/id/eprint/5978/ http://psasir.upm.edu.my/id/eprint/5978/1/FBSB_2006_5.pdf text en public masters Universiti Putra Malaysia Glutathione transferase - Kedah - Cattle Glutathione transferase - Kelantan - Cattle Glutathione transferase - Kedah - Water buffalo Faculty of Biotechnology and Biomokular Sciences English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Glutathione transferase - Kedah - Cattle Glutathione transferase - Kelantan - Cattle Glutathione transferase - Kedah - Water buffalo
spellingShingle	Glutathione transferase - Kedah - Cattle Glutathione transferase - Kelantan - Cattle Glutathione transferase - Kedah - Water buffalo Saleh Huddin, Lailatul Jumaiyah Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver
description	Biotransformation and detoxification process in living organisms consists of two phases, phase I and phase 11. Phase I involves in the introduction of functional group into molecule while the phase I1 involves the conjugation of phase I metabolites. In phase 11, glutathione S-transferases (GSTs; EC 2.5.1.18) has aroused much interest because of its involvement in the biotransformation and detoxification of wide spectrum of xenobiotics which can be from pesticides, herbicides and insecticides. The present study was undertaken to puriG and characterized cytosolic GSTs from livers of Kedah-Kelantan cattle (Bos indicus) and Malaysia water buffalo (Bubalus bubalis). The glutathione Stransferases were isolated from two important livestock livers, Kedah-Kelantan cattle (Bos indicus) and Malaysian water buffalo (Bubalus bubalis) by glutathione affinity chromatography. The affinity-glutathione chromatography successfully purifies the GSTs isoenzymes with 14.73% yield (62.77 purification fold) and 19.71 % yield (20.44 purification fold) for KK cattle and water buffalo livers respectively. Initial methods of purification included centrifugation and ultracentrifugation. The affinity elution with highest activity towards CDNB was estimated for the pI values using isoelectric focusing method via LKB-8 100 ampholyte type (LKB Bromna) apparatus. pI values for affinity purified KK cattle liver are 5.7 (C-34), 6.9 (C-38) and 8.8 (C-42). While for the water buffalo liver, the pI values for glutathione affinity purified isoenzymes are 6.85 (B-23) and 7.2 (B-24). The isoenzymes were then tested using SDS-PAGE method for purity and also to estimate the molecular weight estimation. It has been estimated that molecular weight for water buffalo isoenzymes of B-23 was 29.3 * 0.05 kDa and B-24 was 30.74 * 0.16 kDa. The KK cattle liver isoenzymes molecular weight was estimated with C-34 was 29.9 * 0.14; C-38 was 28.3 * 0.09 and 27.7 *0.03 for C-42. The study showed that KK cattle liver GSTs exist as isoenzymes (PI 8.8, 6.9 and 5.7), and have high activity towards CDNB, low towards DCNB and no activity towards the ethacrynic acid for the substrate specificities. On the other hand, the water buffalo liver GSTs exist as isoenzymes with pI 6.85 and 7.2. For the substrate specificities, the isoenzymes also have high activity for CDNB, but low for DCNB and could not be detected for the ethacrynic acid.
format	Thesis
qualification_level	Master's degree
author	Saleh Huddin, Lailatul Jumaiyah
author_facet	Saleh Huddin, Lailatul Jumaiyah
author_sort	Saleh Huddin, Lailatul Jumaiyah
title	Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver
title_short	Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver
title_full	Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver
title_fullStr	Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver
title_full_unstemmed	Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver
title_sort	partial purification and characterization of glutathione s-transferases from kedah-kelantan cattle (bos indicus) and water buffalo (bubalus bubalis) liver
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomokular Sciences
publishDate	2006
url	http://psasir.upm.edu.my/id/eprint/5978/1/FBSB_2006_5.pdf
_version_	1776100241719689216

Partial Purification and Characterization of Glutathione S-Transferases from Kedah-Kelantan Cattle (Bos Indicus) and Water Buffalo (Bubalus Bubalis) Liver

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