Open AccessOverproduction, purification, crystallization and preliminary X‐ray characterization of a novel carbohydrate‐binding module of endoglucanase Cel5A from Eubacterium cellulosolvens
Author(s) -
Luís Ana S.,
Alves Victor D.,
Romão Maria J.,
Prates José A. M.,
Fontes Carlos M. G. A.,
Najmudin Shabir
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111004246
Subject(s) - cellulase , glycoside hydrolase , hydrolysis , carbohydrate binding module , hydrolase , eubacterium , biochemistry , linker , chemistry , enzyme , clostridium thermocellum , thermophile , stereochemistry , biology , bacteria , genetics , computer science , operating system
The anaerobic cellulolytic rumen bacterium Eubacterium cellulosolvens produces a large array of cellulases and hemicellulases that are responsible for the hydrolysis of plant cell‐wall polysaccharides. One of these enzymes, endoglucanase Cel5A, comprises two tandemly repeated novel carbohydrate‐binding modules (CBMs) and two catalytic domains belonging to glycoside hydrolase family 5 joined by flexible linker sequences. The novel CBM located at the N‐terminus of the endoglucanase has been crystallized. The crystals belonged to the hexagonal space group P 6 1 22 and contained a single molecule in the asymmetric unit. The structure of the l ‐selenomethionine derivative has been solved by a MAD experiment on crystals that diffracted to 1.75 Å resolution.