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Expression, purification and crystallization of a novel carbohydrate‐binding module from the Ruminococcus flavefaciens cellulosome
Author(s) -
Venditto Immacolata,
Centeno Maria S. J.,
Ferreira Luis M. A.,
Fontes Carlos M. G. A.,
Najmudin Shabir
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14024248
Subject(s) - cellulosome , cellulase , carbohydrate binding module , biochemistry , xylanase , ruminococcus , glycoside hydrolase , chemistry , genome , glycobiology , biology , enzyme , computational biology , gene , glycoprotein , clostridium thermocellum , glycan , gut flora
Anaerobic bacteria organize carbohydrate‐active enzymes into a multi‐component complex, the cellulosome, which degrades cellulose and hemicellulose highly efficiently. Genome sequencing of Ruminococcus flavefaciens FD‐1 offers extensive information on the range and diversity of the enzymatic and structural components of the cellulosome. The R. flavefaciens FD‐1 genome encodes over 200 dockerin‐containing proteins, most of which are of unknown function. One of these modular proteins comprises a glycoside hydrolase family 5 catalytic module (GH5) linked to an unclassified carbohydrate‐binding module (CBM‐ Rf 1) and a dockerin. The novel CBM‐ Rf 1 has been purified and crystallized. The crystals belonged to the trigonal space group R 32: H . The CBM‐ Rf 1 structure was determined by a multiple‐wavelength anomalous dispersion experiment using AutoSol from the PHENIX suite using both selenomethionyl‐derivative and native data to resolutions of 2.28 and 2.0 Å, respectively.