Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)

The present study is concerned mainly with partial purification and characterization of cytosolic GST from livers of Malaysian catfish and Malaysian red tilapia as both species are of economic and commercial importance in Malaysia being the major freshwater fishes consumed. This study hopes to e...

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Main Author:	Chin, Chee Meng
Format:	Thesis
Language:	English
Published:	2005
Subjects:	Glutathione transferase - Walking catfish - Malaysia Glutathione transferase - Tilapia - Malaysia
Online Access:	http://psasir.upm.edu.my/id/eprint/5962/1/FBSB_2005_32%20IR.pdf
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id	my-upm-ir.5962
record_format	uketd_dc
spelling	my-upm-ir.59622022-02-17T03:20:05Z Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.) 2005-07 Chin, Chee Meng The present study is concerned mainly with partial purification and characterization of cytosolic GST from livers of Malaysian catfish and Malaysian red tilapia as both species are of economic and commercial importance in Malaysia being the major freshwater fishes consumed. This study hopes to establish the patterns of cytosolic glutathione S-transferase isoenzymes in the catfish and red tilapia. This may be useful to further achieve an understanding toward this enzyme in view of using it as a tool in environmental monitoring. The hepatic GST enzyme from catfish and red tilapia was partially purified 15X and 27X respectively in comparison to the ultra-centrifuged cytosolic fraction by affinity chromatography. Specific GST activity of 12.69 unitlmg protein and 33.42 unitlmg protein was obtained from livers of catfish and red tilapia using l-chloro- 2,4-dinitrobenzene (CDNB) as a substrate. Isolation of GST isoenzymes from affinity purified fractions was achieved by preparative isoelectric-focusing. Two isoenzymes; one major isoenzyme designated Cil and one minor isoenzyme designated Ci2 were isolated from catfish liver with an apparent pl of 6.20 and 8.73 respectively. One isoenzyme designated Ti1 was isolated from red tilapia liver with an apparent pi of 9.14. SDS-PAGE analysis suggests that the isolated isoenzymes appear to be homodimeric in nature with subunit molecular weight of 29.7 21.7 kDa (Cil), 27.7 21.3 kDa (Ci2) and 29.9 +0.9 kDa (Til). As has been found for most GSTs, highest catalytic activity was obtained with CDNB. With the exception of the isoenzyme from cytosol and affinity purified fractions of red tilapia, none of the catfish fractions displayed enzyme activities towards 1,2- dichloro-4-nitrobenzene (DCNB) and ethacrynic acid (EA). Therefore, both catfish and tilapia possess hepatic glutathione S-transferase activity, indicating that they are capable of conjugating endogenous or xenobiotic metabolites/compounds as a result of foreign exposure or oxidative metabolism with glutathione, thereby making it a useful tool as a effective biomarker of aquatic contamination. Glutathione transferase - Walking catfish - Malaysia Glutathione transferase - Tilapia - Malaysia 2005-07 Thesis http://psasir.upm.edu.my/id/eprint/5962/ http://psasir.upm.edu.my/id/eprint/5962/1/FBSB_2005_32%20IR.pdf text en public masters Universiti Putra Malaysia Glutathione transferase - Walking catfish - Malaysia Glutathione transferase - Tilapia - Malaysia Faculty of Biotechnology and Biomokular Sciences Shamaan, Nor Aripin
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
advisor	Shamaan, Nor Aripin
topic	Glutathione transferase - Walking catfish - Malaysia Glutathione transferase - Tilapia - Malaysia
spellingShingle	Glutathione transferase - Walking catfish - Malaysia Glutathione transferase - Tilapia - Malaysia Chin, Chee Meng Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)
description	The present study is concerned mainly with partial purification and characterization of cytosolic GST from livers of Malaysian catfish and Malaysian red tilapia as both species are of economic and commercial importance in Malaysia being the major freshwater fishes consumed. This study hopes to establish the patterns of cytosolic glutathione S-transferase isoenzymes in the catfish and red tilapia. This may be useful to further achieve an understanding toward this enzyme in view of using it as a tool in environmental monitoring. The hepatic GST enzyme from catfish and red tilapia was partially purified 15X and 27X respectively in comparison to the ultra-centrifuged cytosolic fraction by affinity chromatography. Specific GST activity of 12.69 unitlmg protein and 33.42 unitlmg protein was obtained from livers of catfish and red tilapia using l-chloro- 2,4-dinitrobenzene (CDNB) as a substrate. Isolation of GST isoenzymes from affinity purified fractions was achieved by preparative isoelectric-focusing. Two isoenzymes; one major isoenzyme designated Cil and one minor isoenzyme designated Ci2 were isolated from catfish liver with an apparent pl of 6.20 and 8.73 respectively. One isoenzyme designated Ti1 was isolated from red tilapia liver with an apparent pi of 9.14. SDS-PAGE analysis suggests that the isolated isoenzymes appear to be homodimeric in nature with subunit molecular weight of 29.7 21.7 kDa (Cil), 27.7 21.3 kDa (Ci2) and 29.9 +0.9 kDa (Til). As has been found for most GSTs, highest catalytic activity was obtained with CDNB. With the exception of the isoenzyme from cytosol and affinity purified fractions of red tilapia, none of the catfish fractions displayed enzyme activities towards 1,2- dichloro-4-nitrobenzene (DCNB) and ethacrynic acid (EA). Therefore, both catfish and tilapia possess hepatic glutathione S-transferase activity, indicating that they are capable of conjugating endogenous or xenobiotic metabolites/compounds as a result of foreign exposure or oxidative metabolism with glutathione, thereby making it a useful tool as a effective biomarker of aquatic contamination.
format	Thesis
qualification_level	Master's degree
author	Chin, Chee Meng
author_facet	Chin, Chee Meng
author_sort	Chin, Chee Meng
title	Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)
title_short	Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)
title_full	Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)
title_fullStr	Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)
title_full_unstemmed	Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)
title_sort	partial purification and characterization of glutathione s-transferase from the livers of malaysian catfish (clarias batrachus) and malaysian red tilapia (oreochromis sp.)
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomokular Sciences
publishDate	2005
url	http://psasir.upm.edu.my/id/eprint/5962/1/FBSB_2005_32%20IR.pdf
_version_	1747810517266726912

Partial Purification and Characterization of Glutathione S-Transferase from the Livers of Malaysian Catfish (Clarias Batrachus) and Malaysian Red Tilapia (Oreochromis Sp.)

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