Isolation and purification of alkaline phosphatase from porcine intestine using different biochemical techniques

Date

10-2003

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Teresita M. Espino

Co-adviser

Eva Marie Capareda

Committee Member

Hosea Matel

Abstract

Isolation and purification of alkaline phosphatase was conducted using solvent precipitation with n-butanol, ammonium sulfate precipitation (60 and 90% saturation), gel filtration chromatography through Sephadex G-100 and ion-exchange chromatography using DEAE-Sephadex. Alkaline phosphatase activity was determined at each purification step. Increase in enzyme activity was observed at each purification step. Purification folds of 383.34 and 309.94 were obtained for fractions I and 2 respectively after ion-exchange chromatography on DEAE-Sephadex. Physical and chemical properties of purified alkaline phosphatase were determined. The optimum pH for maximum activity for Tris°11CI buffer (1M) was p1-1 8.0 while for diethanolamine-IICI buffer ( I M), pH 9.5. The enzyme was stable at pH 8.0. The optimum temperature in which the enzyme was stable ranged from 5°C to 37°C after 5 minutes incubation. The molecular weight of purified alkaline phosphatase was about 113.50 kDa after SDS-PAGE.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2003 C4 L36

Document Type

Thesis

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