title: Polysaccharide synthesis of the levansucrase SacB from Bacillus megaterium is controlled by distinct surface motifs.
abstract: Despite the widespread biological function of carbohydrates, the polysaccharide synthesis mechanisms of glycosyltransferases remain largely unexplored. Bacterial levansucrases (glycoside hydrolase family 68) synthesize high molecular weight, β-(2,6)-linked levan from sucrose by transfer of fructosyl units. The kinetic and biochemical characterization of Bacillus megaterium levansucrase SacB variants Y247A, Y247W, N252A, D257A, and K373A reveal novel surface motifs remote from the sucrose binding site with distinct influence on the polysaccharide product spectrum. The wild type activity (k(cat)) and substrate affinity (K(m)) are maintained. The structures of the SacB variants reveal clearly distinguishable subsites for polysaccharide synthesis as well as an intact active site architecture. These results lead to a new understanding of polysaccharide synthesis mechanisms. The identified surface motifs are discussed in the context of related glycosyltransferases.
authors: Department of Molecular Structural Biology, Helmholtz-Centre for Infection Research, Inhoffenstrasse 7B, 38124 Braunschweig, Germany.
authors: Christian P
authors: CP
authors: Strube
authors: Arne
authors: A
authors: Homann
authors: Martin
authors: M
authors: Gamer
authors: Dieter
authors: D
authors: Jahn
authors: Jürgen
authors: J
authors: Seibel
authors: Dirk W
authors: DW
authors: Heinz
mesh_terms: D020816:Amino Acid Motifs; D019943:Amino Acid Substitution; D001410:Bacillus megaterium / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006602:Hexosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020125:Mutation, Missense; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011135:Polysaccharides, Bacterial; D006602:Hexosyltransferases; C021513:levansucrase