title: Cel5I, a SLH-Containing Glycoside Hydrolase: Characterization and Investigation on Its Role in Ruminiclostridium cellulolyticum.
abstract: Ruminiclostridium cellulolyticum (Clostridium cellulolyticum) is a mesophilic cellulolytic anaerobic bacterium that produces a multi-enzymatic system composed of cellulosomes and non-cellulosomal enzymes to degrade plant cell wall polysaccharides. We characterized one of the non-cellulosomal enzymes, Cel5I, composed of a Family-5 Glycoside Hydrolase catalytic module (GH5), a tandem of Family-17 and -28 Carbohydrate Binding Modules (CBM), and three S-layer homologous (SLH) modules, where the latter are expected to anchor the protein on the cell surface. Cel5I is the only putative endoglucanase targeting the cell surface as well as the only putative protein in R. cellulolyticum containing CBM17 and/or CBM28 modules. We characterized different recombinant structural variants from Cel5I. We showed that Cel5I has an affinity for insoluble cellulosic substrates through its CBMs, that it is the most active endoglucanase on crystalline cellulose of R. cellulolyticum characterized to date and mostly localized in the cell envelope of R. cellulolyticum. Its role in vivo was analyzed using a R. cellulolyticum cel5I mutant strain. Absence of Cel5I in the cell envelope did not lead to a significant variation of the phenotype compared to the wild type strain. Neither in terms of cell binding to cellulose, nor for its growth on crystalline cellulose, thus indicating that the protein has a rather subtle role in tested conditions. Cel5I might be more important in a natural environment, at low concentration of degradable glucose polymers, where its role might be to generate higher concentration of short cellodextrins close to the cell surface, facilitating their uptake or for signalization purpose.
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Nathalie
authors: N
authors: Franche
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Chantal
authors: C
authors: Tardif
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Julie
authors: J
authors: Ravachol
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Seddik
authors: S
authors: Harchouni
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Pierre-Henri
authors: PH
authors: Ferdinand
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Romain
authors: R
authors: Borne
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Henri-Pierre
authors: HP
authors: Fierobe
authors: Aix Marseille Univ, CNRS, LCB, Marseille, France.
authors: Stéphanie
authors: S
authors: Perret
mesh_terms: D050260:Carbohydrate Metabolism; D002480:Cellulase / Q000378:metabolism*; D044603:Cellulosomes / Q000378:metabolism*; D046973:Ruminiclostridium cellulolyticum / Q000201:enzymology*; D006027:Glycosides / Q000378:metabolism*; D006868:Hydrolysis
chemical_list: D006027:Glycosides; D002480:Cellulase