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Cloning and molecular modelling of pectin degrading glycosyl hydrolase of family 28 from soil metagenomic library.

Sathya, T. A. and Ani, Jacob Mathew and Mahejibin, Khan (2014) Cloning and molecular modelling of pectin degrading glycosyl hydrolase of family 28 from soil metagenomic library. Molecular Biology Reports, 41. pp. 2645-2656.

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Abstract

Western Ghats of India is recognized as one of the 12 mega diversity regions of the world and is the hot spot for unrevealed microbial diversity. To explore the diversity of polysaccharide degrading enzymes in that region, metagenomic library was constructed from forest soil of Southern Western Ghats region. Nine pectinolytic clones with the ability to degrade citrus pectin were isolated based on function based screening of the library. Sequence analysis of pg_4 clone containing revealed that it contained GH family 28 domain (pfam00295) belonging to polygalacturonase superfamily (PLN03003). Its amino acid sequence analysis showed 25–55 % identity to the other well-characterized polygalacturonases. Molecular modeling of pg_4 revealed that it comprised of three right handed-parallel b sheets, one anti-parallel b sheet and one a helix with three conserved catalytic residue D 2263, D 284-85 and H 312 at the C terminal end. The enzyme characterized was able to hydrolyze both apple and citrus pectin with Km values of 1.685 and 1.542 mg ml-1 and retained more that 80 % of activity at pH 5–9 and temperature 20–60 �C.

Item Type: Article
Uncontrolled Keywords: Glycosyl hydrolase Metagenomics Polygalacturonase
Subjects: 500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 28 Polysaccharide Chemistry
Divisions: Food Microbiology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 25 Apr 2014 10:12
Last Modified: 18 Nov 2016 08:19
URI: http://ir.cftri.com/id/eprint/11469

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