Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)

Alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, which catalyze nonspecific hydrolysis of phosphate monoesters. Partial purification was conducted on alkaline phosphatase (ALP) extracted from hepatopancreas and intestine of red tilapia, (Tilapia mossombica) using two main steps - ammonium s...

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Main Author:	Mariappan, Vanitha
Format:	Thesis
Language:	English
Published:	2005
Subjects:	Mozambique tilapia
Online Access:	http://psasir.upm.edu.my/id/eprint/5957/1/FBSB_2005_30%20IR.pdf
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id	my-upm-ir.5957
record_format	uketd_dc
spelling	my-upm-ir.59572022-02-17T03:10:19Z Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica) 2005-07 Mariappan, Vanitha Alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, which catalyze nonspecific hydrolysis of phosphate monoesters. Partial purification was conducted on alkaline phosphatase (ALP) extracted from hepatopancreas and intestine of red tilapia, (Tilapia mossombica) using two main steps - ammonium sulphate precipitation and ion exchange chromatography on DEAE - 52. Samples from the ion-exchange step were analysed for ALP activities and characterised by SDS-PAGE. SDS-PAGE analysis showed 2 identical bands and was found to have molecular weight of 68,000 Da (hepatopancreas ALP) and 180,500 Da (intestinal ALP) subunits. Overall, purification fold obtained from the final step are 1.8 and 21.9 for hepatopancreas and intestinal respectively, with recovery of only 0.22% from hepatopancreas and 0.01% from intestine. The specific activity of the enzyme was 1.72 X 10- 2 pmol min-1 mg-1 and 2.93 X 10-1 pmol min-1 mg-1 from hepatopancreas and intestine respectively. The ALP from hepatopancreas remained stable at temperatures up to 50°C, and ALP from intestine enzyme had an optimum temperature of 60°C. The optimum pH for both hepatopancreas and intestine ALP of Tilapia mossambica is pH 10. The positive monovalent alkali metal ions (Li+, Na+ and K+) have no effect on the ALP enzyme activity. However, the positive divalent alkali metal ions (Mg2+a nd Ca2+)a ctivate the enzyme activities. Heavy metal ions (Zn2+, CU~+C,d 2+a nd Hg2+) were found to inhibit the enzyme activity. Mozambique tilapia 2005-07 Thesis http://psasir.upm.edu.my/id/eprint/5957/ http://psasir.upm.edu.my/id/eprint/5957/1/FBSB_2005_30%20IR.pdf text en public masters Universiti Putra Malaysia Mozambique tilapia Faculty of Biotechnology and Biomokular Sciences Shamaan, Nor Aripin
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
advisor	Shamaan, Nor Aripin
topic	Mozambique tilapia
spellingShingle	Mozambique tilapia Mariappan, Vanitha Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)
description	Alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, which catalyze nonspecific hydrolysis of phosphate monoesters. Partial purification was conducted on alkaline phosphatase (ALP) extracted from hepatopancreas and intestine of red tilapia, (Tilapia mossombica) using two main steps - ammonium sulphate precipitation and ion exchange chromatography on DEAE - 52. Samples from the ion-exchange step were analysed for ALP activities and characterised by SDS-PAGE. SDS-PAGE analysis showed 2 identical bands and was found to have molecular weight of 68,000 Da (hepatopancreas ALP) and 180,500 Da (intestinal ALP) subunits. Overall, purification fold obtained from the final step are 1.8 and 21.9 for hepatopancreas and intestinal respectively, with recovery of only 0.22% from hepatopancreas and 0.01% from intestine. The specific activity of the enzyme was 1.72 X 10- 2 pmol min-1 mg-1 and 2.93 X 10-1 pmol min-1 mg-1 from hepatopancreas and intestine respectively. The ALP from hepatopancreas remained stable at temperatures up to 50°C, and ALP from intestine enzyme had an optimum temperature of 60°C. The optimum pH for both hepatopancreas and intestine ALP of Tilapia mossambica is pH 10. The positive monovalent alkali metal ions (Li+, Na+ and K+) have no effect on the ALP enzyme activity. However, the positive divalent alkali metal ions (Mg2+a nd Ca2+)a ctivate the enzyme activities. Heavy metal ions (Zn2+, CU~+C,d 2+a nd Hg2+) were found to inhibit the enzyme activity.
format	Thesis
qualification_level	Master's degree
author	Mariappan, Vanitha
author_facet	Mariappan, Vanitha
author_sort	Mariappan, Vanitha
title	Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)
title_short	Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)
title_full	Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)
title_fullStr	Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)
title_full_unstemmed	Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)
title_sort	partial purification and characterisation of alkaline phosphatase from hepatopancreas and intertine of red tilapia, (tilapia mossambica)
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomokular Sciences
publishDate	2005
url	http://psasir.upm.edu.my/id/eprint/5957/1/FBSB_2005_30%20IR.pdf
_version_	1747810516300988416

Partial Purification and Characterisation of Alkaline Phosphatase from Hepatopancreas and Intertine of Red Tilapia, (Tilapia Mossambica)

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