title: Expression, purification and crystallization of a novel carbohydrate-binding module from the Ruminococcus flavefaciens cellulosome.
abstract: 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-Rf1) and a dockerin. The novel CBM-Rf1 has been purified and crystallized. The crystals belonged to the trigonal space group R32:H. The CBM-Rf1 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.
authors.affiliation: CIISA, Faculdade de Medicina Veterinaria, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Immacolata
authors.initials: I
authors.lastname: Venditto
authors.affiliation: CIISA, Faculdade de Medicina Veterinaria, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Maria S J
authors.initials: MS
authors.lastname: Centeno
authors.affiliation: CIISA, Faculdade de Medicina Veterinaria, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Luis M A
authors.initials: LM
authors.lastname: Ferreira
authors.affiliation: CIISA, Faculdade de Medicina Veterinaria, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Carlos M G A
authors.initials: CM
authors.lastname: Fontes
authors.affiliation: CIISA, Faculdade de Medicina Veterinaria, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisbon, Portugal.
authors.forename: Shabir
authors.initials: S
authors.lastname: Najmudin
mesh_terms: D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000737:chemistry*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D045857:Ruminococcus / Q000737:chemistry*
chemical_list: D002241:Carbohydrates
keywords: Ruminococcus flavefaciens; cellulosome; glycoside hydrolase family 5 catalytic module; novel carbohydrate-binding module