Isolation and purification of bromelain from pineapple wastes

Date

4-2003

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Teresita M. Espino

Co-adviser

Florinia Merca

Committee Member

Macario Catahan

Abstract

Bromelain from pineapple wastes (crown, peel, eyes and core of pineapple) was isolated and purified. Purification of the enzyme involved ammonium sulfate precipitation, desalting by gel filtration on Sephadex G-25 column, ion exchange chromatography on DEAF-cellulose column, and gel filtration on Sephadex G-100 column. Increase in specific activity was observed at each purification step except in gel filtration on Sephadex G-100 due to freeze drying. The isolated bromelain was purified 16.91 times that of the crude enzyme extract alter ion exchange chromatography on DEAF-cellulose and 6.34 times after gel filtration on Sephadex G-100. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the purified bromelain preparation contained two distinct enzymes with molecular weights of 16.89 kDa and 18.95 kDa. Non-denaturing gel electrophoresis and gel filtration chromatography on Sephadex (i-100 Superfine confirmed the heterogeneity of the preparation. Characterization of the partially purified prom lain showed optimum proteolytic activity at 60°C, and at pl 1 5.0 and 9.0 using azocasein as substrate. Bromelain was stable over a wide range of pH (4.0 to 10.0). It was unstable at 20 to 70°C after incubation at these temperatures for an hour.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2003 C4 I43

Document Type

Thesis

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