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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/13325
Title: Purification, bio-chemical characterization, homology modeling and active site binding mode interactions of thermo-alkali-tolerant β-1,4 endoxylanase from Coprinus cinereus LK-D-NCIM-1369
Authors: Kumar L.
Dutt D.
Tapas S.
Kumar P.
Published in: Biocatalysis and Agricultural Biotechnology
Abstract: Present study aims at purifying and characterizing crude xylanase isolated from Coprinus cinereus LK-D-NCIM-1369 under solid-state fermentation conditions. Crude xylanase is purified by (NH4)2SO4 precipitation, carboxymethyl cellulose cation-exchange and gel filtration chromatography with Superdex-200 column. Purified xylanase from C. cinereus shows 54.3% similarity with β-1,4 endoxylanase from C. cinerea okayama7#130 with accession number gi|169855830 based on MALDI-TOF/TOF. The purified xylanase-a monomeric protein with molecular weight 20.1kD shows maximum activity at 60°C and pH 7.0 and stable over pH 5.0-9.0 and temperature up to 70°C. The xylanase shows Km and V values of 3.26mg/mL and 909.09μm/min/mg for birchwood xylan. A sequence of 186 amino acids of C. cinerea okayama7#130 gi|169855830 is retrieved from NCBI database and its 3-D model is generated on the basis of crystal structure of xylanase 1XNK-A from Chaetomium thermophilum with the help of online server SWISS-MODEL workspace. The model is verified and validated on SAVES and PROCHECK programmes, respectively. Ramachandran plot reveals that the total residues in allowed and generously allowed regions are 99.4% and 0.6% respectively. Overlapping of xylanase with the template of 1XNK-A stipulates the amino acid residues Asn35, Tyr68, Arg113, Ser118, Tyr168 and Glu174 constitute active site of the enzyme. © 2013 Elsevier Ltd.
Citation: Biocatalysis and Agricultural Biotechnology (2013), 2(3): 267-277
URI: https://doi.org/10.1016/j.bcab.2013.04.004
http://repository.iitr.ac.in/handle/123456789/13325
Issue Date: 2013
Keywords: Docking
Homology modeling
MALDI-TOF/TOF
Purification
Thermo-pH-stability
Xylanase
ISSN: 18788181
Author Scopus IDs: 57208572291
7005982560
35491918300
55064809000
Author Affiliations: Kumar, L., Department of Paper Technology, Indian Institute of Technology Roorkee, Saharanpur Campus, Saharanpur 247001, India
Dutt, D., Department of Paper Technology, Indian Institute of Technology Roorkee, Saharanpur Campus, Saharanpur 247001, India
Tapas, S., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Kumar, P., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Funding Details: The first author is thankful to the Council of Scientific and Industrial Research , New Delhi for awarding him Senior Research Fellowship and for providing partial financial support to carry out this work.
Corresponding Author: Dutt, D.; Department of Paper Technology, Indian Institute of Technology Roorkee, Saharanpur Campus, Saharanpur 247001, India; email: dharmduttiit@gmail.com
Appears in Collections:Journal Publications [PT]

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