Open Access Journal

ISSN : 2456-1304 (Online)

International Journal of Engineering Research in Electronics and Communication Engineering(IJERECE)

Monthly Journal for Electronics and Communication Engineering

Open Access Journal

International Journal of Science Engineering and Management (IJSEM)

Monthly Journal for Science Engineering and Management

ISSN : 2456-1304 (Online)

Isolation, Screening and Molecular Characterization of Polygalacturonase Enzyme from Bacillus Species using RAPD Markers

Author : Dhananjaiah T M 1 Dr.Prasad.M.P 2

Date of Publication :15th July 2020

Abstract: Pectic substances form the major components of the plant cell wall. Polygalacturonase is a hydrolytic enzyme, which acts on polygalacturonic acid (PGA), hydrolyzing α-1,4 glycosidic bonds of pectic acid cleaving the α (1-4) bonds between adjacent galacturonic acid within the homogalacturonic acid backbone of pectin. Pectin modifying enzymes are formed in huge amount by plant related Microbes. Polygalacturonase enzyme has vital role in extraction and softening of fruits, and also plays a major role in agriculture and environmental sectors. The potential of using microorganisms as a biotechnological source in the production of industrially relevant food and textile processing enzymes, has stimulated renewed interest in the exploration of extracellular enzymatic activity. Production and scale up of Bacterial enzymes are easy, low cost and can act on a wider range of pH and environment. The current research was aimed at isolating pectinolytic Bacillus sp. from municipal dump sites, screen for polygalacturonase enzyme production and to determine the genetic variability. Six different soil samples were collected and plated on nutrient agar and the isolated colonies were screened for polygalacturonase production on modified MS medium. Of the 20 isolates screened, 8 isolated exhibited clear zones of hydrolysis indicating positive activity for polygalacturonase enzyme. The organisms were identified as Bacillus sp. based on colony, cell and biochemical characterization. The genetic variability of these polygalacturonase enzyme producing Bacillus sp. were analyzed using PCR based RAPD technique using four primers RBA1, RBA2, RBA3, RBA4. Three different groups of RAPD profiles were determined within the eight species based on the distance calculated by Euclidean Similarity matrix. The First clade has 4 organisms with ORG13 and ORG18 closely related followed by ORG1 and ORG15 and in the second clade ORG4 and ORG12 are closely related and according to the third clade ORG16 and ORG17 were found to have close evolutionary relationship. The genetic diversity is evident based on the percentage similarity between the isolates.

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