title: Characterization and crystal structure determination of β-1,2-mannobiose phosphorylase from Listeria innocua.
abstract: Glycoside hydrolase family 130 consists of phosphorylases and hydrolases for β-mannosides. Here, we characterized β-1,2-mannobiose phosphorylase from Listeria innocua (Lin0857) and determined its crystal structures complexed with β-1,2-linked mannooligosaccharides. β-1,2-Mannotriose was bound in a U-shape, interacting with a phosphate analog at both ends. Lin0857 has a unique dimer structure connected by a loop, and a significant open-close loop displacement was observed for substrate entry. A long loop, which is exclusively present in Lin0857, covers the active site to limit the pocket size. A structural basis for substrate recognition and phosphorolysis was provided.
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Tsuda
authors.affiliation: Faculty of Agriculture, Niigata University, Niigata 950-2181, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Nihira
authors.affiliation: Faculty of Agriculture, Niigata University, Niigata 950-2181, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Chiku
authors.affiliation: Faculty of Agriculture, Niigata University, Niigata 950-2181, Japan.
authors.forename: Erika
authors.initials: E
authors.lastname: Suzuki
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Takatoshi
authors.initials: T
authors.lastname: Arakawa
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Nishimoto
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Motomitsu
authors.initials: M
authors.lastname: Kitaoka
authors.affiliation: Faculty of Agriculture, Niigata University, Niigata 950-2181, Japan. Electronic address: nakai@agr.niigata-u.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakai
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: asfushi@mail.ecc.u-tokyo.ac.jp.
authors.forename: Shinya
authors.initials: S
authors.lastname: Fushinobu
mesh_terms: D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D008087:Listeria / Q000201:enzymology*; D008351:Mannans / Q000378:metabolism*; D008958:Models, Molecular; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D006005:Phosphorylases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D008351:Mannans; D009844:Oligosaccharides; C014647:mannobiose; D006005:Phosphorylases
keywords: Enzyme–substrate interaction; Glycoside hydrolase family 130; Oligosaccharide synthesis; Structure–function relationship; Substrate specificity; β-1,2-Mannobiose phosphorylase