Polysaccharide synthesis of the levansucrase SacB from Bacillus megaterium is controlled by distinct surface motifs.

2.50
Hdl Handle:
http://hdl.handle.net/10033/202369
Title:
Polysaccharide synthesis of the levansucrase SacB from Bacillus megaterium is controlled by distinct surface motifs.
Authors:
Strube, Christian P; Homann, Arne; Gamer, Martin; Jahn, Dieter; Seibel, Jürgen; Heinz, Dirk W
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.
Affiliation:
Department of Molecular Structural Biology, Helmholtz-Centre for Infection Research, Inhoffenstrasse 7B, 38124 Braunschweig, Germany.
Citation:
Polysaccharide synthesis of the levansucrase SacB from Bacillus megaterium is controlled by distinct surface motifs. 2011, 286 (20):17593-600 J. Biol. Chem.
Journal:
The Journal of biological chemistry
Issue Date:
20-May-2011
URI:
http://hdl.handle.net/10033/202369
DOI:
10.1074/jbc.M110.203166
PubMed ID:
21454585
Type:
Article
Language:
en
ISSN:
1083-351X
Appears in Collections:
Publications from Division of Molekulare Struktur Biologie (MOSB)

Full metadata record

DC FieldValue Language
dc.contributor.authorStrube, Christian Pen
dc.contributor.authorHomann, Arneen
dc.contributor.authorGamer, Martinen
dc.contributor.authorJahn, Dieteren
dc.contributor.authorSeibel, Jürgenen
dc.contributor.authorHeinz, Dirk Wen
dc.date.accessioned2012-01-11T13:03:17Z-
dc.date.available2012-01-11T13:03:17Z-
dc.date.issued2011-05-20-
dc.identifier.citationPolysaccharide synthesis of the levansucrase SacB from Bacillus megaterium is controlled by distinct surface motifs. 2011, 286 (20):17593-600 J. Biol. Chem.en
dc.identifier.issn1083-351X-
dc.identifier.pmid21454585-
dc.identifier.doi10.1074/jbc.M110.203166-
dc.identifier.urihttp://hdl.handle.net/10033/202369-
dc.description.abstractDespite 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.en
dc.language.isoenen
dc.subject.meshAmino Acid Motifsen
dc.subject.meshAmino Acid Substitutionen
dc.subject.meshBacillus megateriumen
dc.subject.meshBacterial Proteinsen
dc.subject.meshHexosyltransferasesen
dc.subject.meshMutation, Missenseen
dc.subject.meshPolysaccharides, Bacterialen
dc.titlePolysaccharide synthesis of the levansucrase SacB from Bacillus megaterium is controlled by distinct surface motifs.en
dc.typeArticleen
dc.contributor.departmentDepartment of Molecular Structural Biology, Helmholtz-Centre for Infection Research, Inhoffenstrasse 7B, 38124 Braunschweig, Germany.en
dc.identifier.journalThe Journal of biological chemistryen

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