title: A noncellulosomal mannanase26E contains a CBM59 in Clostridium cellulovorans.
abstract: A multicomponent enzyme-complex prevents efficient degradation of the plant cell wall for biorefinery. In this study, the method of identifying glycoside hydrolases (GHs) to degrade hemicelluloses was demonstrated. The competence of C. cellulovorans, which changes to be suitable for degradation of each carbon source, was used for the method. C. cellulovorans was cultivated into locust bean gum (LBG) that is composed of galactomannan. The proteins produced by C. cellulovorans were separated into either fractions binding to crystalline cellulose or not. Proteins obtained from each fraction were further separated by SDS-PAGE and were stained with Coomassie Brilliant Blue and were detected for mannanase activity. The proteins having the enzymatic activity for LBG were cut out and were identified by mass spectrometry. As a result, four protein bands were classified into glycosyl hydrolase family 26 (GH26) mannanases. One of the identified mannanases, Man26E, contains a carbohydrate-binding module (CBM) family 59, which binds to xylan, mannan, and Avicel. Although mannose and galactose are the same as a hexose, the expression patterns of the proteins from C. cellulovorans were quite different. More interestingly, zymogram for mannanase activity showed that Man26E was detected in only LBG medium.
authors.affiliation: Department of Life Sciences, Graduate School of Bioresources, Mie University, 1577 Kurimamachiya, Tsu, Mie 514-8507, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Department of Life Sciences, Graduate School of Bioresources, Mie University, 1577 Kurimamachiya, Tsu, Mie 514-8507, Japan ; Graduate School of Bioresources, Department of Bioinfomatics, Mie University Life Science Research Center, Mie University, 1577 Kurimamachiya, Tsu, Mie 514-8507, Japan ; Laboratory of Applied Biotechnology, Mie University Industrial Technology Innovation Institute, Mie University, 1577 Kurimamachiya, Tsu, Mie 514-8507, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Tamaru
mesh_terms: D002480:Cellulase / Q000737:chemistry* / Q000302:isolation & purification; D046974:Clostridium cellulovorans / Q000201:enzymology*; D004789:Enzyme Activation; D005685:Galactans / Q000378:metabolism*; D008351:Mannans / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification; D053149:Plant Gums / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry*; D013379:Substrate Specificity; D044902:beta-Mannosidase / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D005685:Galactans; D008351:Mannans; D009097:Multienzyme Complexes; D053149:Plant Gums; D011134:Polysaccharides; C473132:cellulosome, Clostridium; C007916:hemicellulose; D044902:beta-Mannosidase; D002480:Cellulase; C017471:locust bean gum