title: Overproduction, purification, crystallization and preliminary X-ray characterization of a novel carbohydrate-binding module of endoglucanase Cel5A from Eubacterium cellulosolvens.
abstract: 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 P6(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.
authors.affiliation: CIISA - Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal.
authors.forename: Ana S
authors.initials: AS
authors.lastname: Luís
authors.forename: Victor D
authors.initials: VD
authors.lastname: Alves
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Romão
authors.forename: José A M
authors.initials: JA
authors.lastname: Prates
authors.forename: Carlos M G A
authors.initials: CM
authors.lastname: Fontes
authors.forename: Shabir
authors.initials: S
authors.lastname: Najmudin
mesh_terms: D050260:Carbohydrate Metabolism; D002480:Cellulase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D005051:Eubacterium / Q000201:enzymology*; D011485:Protein Binding
chemical_list: C407024:endoglucanase Cel5A; D002480:Cellulase