ISOLATION, MOLECULAR CHARACTERISATION AND IMPROVEMENT OF PECTINASE GENE(S) FROM LOCAL BACTERIAL ISOLATES

Abstract

In this study, twenty two bacterial isolates were isolated from agricultural crop waste soil at National Research Center. After optimization conditions using different parameters to test pectinase specific activity for high seven and two leaky isolates. The maximum pectinase specific active isolates were No. 2, 7 and 22 which recorded 71.10, 79.98 and 91.45 U/ml, respectively. Biochemical identification depended on morphological, physiological and biochemical parameters for these isolates No.2, 7 and 22 was confirmed with molecular identification by PCR amplification and sequencing of 16S rRNA gene, identified as; Bacillus filamentosus N2, Bacillus endophyticus N7 and Klebsiella oxytoca N22.These most three productive identified strains were treated in third step mutagenesis with ethedium bromide (Eth.Br.) and hydrogen peroxide (H2O2) for creating super productive mutants.