Gupta, Puneet and Mishra, Arjun K. and Vakhlu, Jyoti (2017) Cloning and characterization of thermo-alkalistable and surfactant stable endoglucanase from Puga hot spring metagenome of Ladakh (J&K). International Journal of Biological Macromolecules, 103. pp. 870-877. ISSN 01418130

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Text Cloning and characterization of thermo-alkalistable and surfactant stable endoglucanase from Puga hot spring metagenome of Ladakh (J&K).pdf Download (2434Kb) | Preview

Abstract

A thermo-alkalistable and surfactant stable endoglucanase (PHS) gene consisting of 554 amino acidswas identified from metagenomic library of Puga hot spring using functional screening. PHS gene wasoverexpressed and purified to homogeneity using affinity chromatography The purified PHS proteinpresented a single band of 60 kDa on the SDS-PAGE gel and zymogram. The recombinant PHS exhibitedactivity over a broad range of pH and temperature with optima at pH 8.0 and 65◦C, respectively and havingoptimum stability at 60◦C and pH 8.0, respectively. The recombinant PHS showed highest substratespecificity using CMC (218.4 U/mg) as compared with Barley �-glucan (89.2 U/mg) and Avicel (0.8U/mg).The Kmand Vmaxof recombinant PHS for CMC were 3.85 mg/ml and 370.37 �molmin−1mg−1, respectively.The activity of the recombinant PHS was enhanced by treatment with 10 mM non-ionic detergents suchas Tween 20, Tween 40, Tween 80, Triton X- 100 and PEG and was inhibited by CTAB, SDS. Its functionalitywas stable in the presence of Fe3+but inhibited by Cu2+, Hg2+, Mn2+and Zn2+. These properties make PHSendoglucanase a potential candidate for use in laundry, textile,paper and pulp industries

Item Type:	Article
Subjects:	Biochemical and Biophysical Sciences
Depositing User:	RCB Library
Date Deposited:	17 Jul 2020 11:43
Last Modified:	17 Jul 2020 11:43
URI:	http://rcb.sciencecentral.in/id/eprint/370

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