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Screening And Characterisation Of Pectinase And Antimicrobial Compounds From Soil Metagenomic Library

Subanthini, A. (2011) Screening And Characterisation Of Pectinase And Antimicrobial Compounds From Soil Metagenomic Library. [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type: Student Project Report
Additional Information: The aim of the present study is to screen polygalactouronase producing clone from soil metagenomic library. Therefore a total of nine clones were randomly selected and were screened from soil metagenomic library. Clones of RB4, RB5, RB8, RB15, were selected, among which the clone RB8 was found to produce significant amount of polygalacturonase. The pectinase produced by RB8 clone was purified by ammonium sulphate precipitation and size exclusion chromatography. Molecular weight of partially purified enzyme was found to be 66kDa based on SDS-PAGE and further confirmed by zymography study. Under partially optimized conditions, purified polygalacturonase showed enzyme activity of 125.46U/ml and protein concentration of 104.78μg/ml at pH 6.0 and temperature 40°C.The enzyme was comparatively stable from 20°C to 40°C and the activity decreased with increasing and decreasing the temperature. In view of cost effective pectinase production, substrate optimization using soild state fermentation was carried out for rotten apple pulp and orange baggase, amongst which rotten apple pulp gave 1241.44U/ml polygalacturonase activity which was higher than 655.41U/ml of orange bagasse. Apart from this, a trial was done to find a antimicrobial agent producing clones from soil metagenomic library. Five clones were screened for antimicrobial activity against six pathogenic strains obtained from MTCC-(IMTECH, Chandigarh). Among the five soil metagenomic clones, only three (RB8, RB15 and S9) showed better inhibition against two pathogenic strains. These clones were capable of biosynthesizing antimicrobial metabolites. Further separation and characterization of active metabolites using TLC- Bioautography were performed.
Uncontrolled Keywords: polygalactouronase, soil metagenomic library, antimicrobial metabolites
Subjects: 500 Natural Sciences and Mathematics > 05 Earth Sciences > 02 Soil Sciences
Divisions: Food Microbiology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 05 Jul 2011 07:22
Last Modified: 28 Dec 2011 10:25
URI: http://ir.cftri.com/id/eprint/10192

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