"Purification of pectinase by dye -ligand chromatography"

Date

3-1999

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Myrna S. Rodriguez

Abstract

PADOLINA, LINGLINGAY D., B.S. Chemistry, University of the Philippines at Los Banos, March 1998, "Purification of Pectinase by Dye-Ligand Chromatography".

Adviser: Dr. Myrna S. Rodriguez

Aspergillus sp. (strain number: Hennings Cal CL 21; BIOTECH accession number: 3128), a fungal isolate was used as source of the pectic enzymes. The production of pectic enzymes in enameled trays was done using the optimized conditions for solid substrate cultivation.

Twenty dyes were screened to choose the positive- and the negative-binding dyes. Dye no. 15 and 78 were chosen as negative and positive dyes, respectively. However, dye no. 15 or Brown MX-GRN proved to be an ineffective negative-binding dye adsorbent on a large scale since it retains more than 90% of the enzyme when it should have not adsorbed it if it were acting as a negative column. Elution with 0.02 M NaCl proved to be effective in desorbing the enzyme from the dye because the last eluate collected had a specific activity for the eluate of 0.98 units PG/mg protein.

Elution with 0.0001 M galacturonic acid which is the substrate was also effective in elution because the last eluate collected had a specific activity for the eluate of 1.07 units PG/mg protein.

The recommended dye to be used for the purification of pectinase is dye no 78 and the recommended method of elution is by increasing the ionic strength using 0.02 M NaCI.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 1999 C4 P33

Document Type

Thesis

This document is currently not available here.

Share

COinS