Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/90026
Citations
Scopus Web of Science® Altmetric
?
?
Type: Journal article
Title: BcsA and BcsB form the catalytically active core of bacterial cellulose synthase sufficient for in vitro cellulose synthesis
Author: Omadjela, O.
Narahari, A.
Strumillo, J.
Mélida, H.
Mazur, O.
Bulone, V.
Zimmer, J.
Citation: Proceedings of the National Academy of Sciences of the United States of America, 2013; 110(44):17856-17861
Publisher: National Academy of Sciences
Issue Date: 2013
ISSN: 0027-8424
1091-6490
Statement of
Responsibility: 
Okako Omadjela, Adishesh Narahari, Joanna Strumillo, Hugo Mélida, Olga Mazur, Vincent Bulone, and Jochen Zimmer
Abstract: Cellulose is a linear extracellular polysaccharide. It is synthesized by membrane-embedded glycosyltransferases that processively polymerize UDP-activated glucose. Polymer synthesis is coupled to membrane translocation through a channel formed by the cellulose synthase. Although eukaryotic cellulose synthases function in macromolecular complexes containing several different enzyme isoforms, prokaryotic synthases associate with additional subunits to bridge the periplasm and the outer membrane. In bacteria, cellulose synthesis and translocation is catalyzed by the inner membrane-associated bacterial cellulose synthase (Bcs)A and BcsB subunits. Similar to alginate and poly-β-1,6 N-acetylglucosamine, bacterial cellulose is implicated in the formation of sessile bacterial communities, termed biofilms, and its synthesis is likewise stimulated by cyclic-di-GMP. Biochemical studies of exopolysaccharide synthesis are hampered by difficulties in purifying and reconstituting functional enzymes. We demonstrate robust in vitro cellulose synthesis reconstituted from purified BcsA and BcsB proteins from Rhodobacter sphaeroides. Although BcsA is the catalytically active subunit, the membrane-anchored BcsB subunit is essential for catalysis. The purified BcsA-B complex produces cellulose chains of a degree of polymerization in the range 200-300. Catalytic activity critically depends on the presence of the allosteric activator cyclic-di-GMP, but is independent of lipid-linked reactants. Our data reveal feedback inhibition of cellulose synthase by UDP but not by the accumulating cellulose polymer and highlight the strict substrate specificity of cellulose synthase for UDP-glucose. A truncation analysis of BcsB localizes the region required for activity of BcsA within its C-terminal membrane-associated domain. The reconstituted reaction provides a foundation for the synthesis of biofilm exopolysaccharides, as well as its activation by cyclic-di-GMP.
Keywords: Biofilms; Escherichia coli; Rhodobacter sphaeroides; Cellulose; Glucosyltransferases; Protein Subunits; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Biological Transport; Gas Chromatography-Mass Spectrometry
Rights: Copyright status unknown
RMID: 0030021021
DOI: 10.1073/pnas.1314063110
Appears in Collections:Agriculture, Food and Wine publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.