Glycobiology Advance Access originally published online on July 31, 2006
Glycobiology 2006 16(11):1021-1032; doi:10.1093/glycob/cwl029
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Cloning and biochemical characterization of the fucanase FcnA: definition of a novel glycoside hydrolase family specific for sulfated fucans
2 Equipe Glycobiologie Marine, Centre National de la Recherche Scientifique, Université Pierre et Marie Curie-Paris6, Unité Mixte de Recherche 7139, Station Biologique, F-29682 Roscoff Cedex, Bretagne, France; 3 Service Commun de Résonance Magnétique Nucléaire, Université de Bretagne Occidentale, Brest, Bretagne, France; and 4 Laboratoires Goëmar, Saint Malo, Bretagne, France
1 To whom correspondence should be addressed; e-mail: gurvan{at}sb-roscoff.fr
Received on December 5, 2005; revised on June 23, 2006; accepted on July 18, 2006
Sulfated fucans are matrix polysaccharides from marine brown algae, consisting of an 
-L-fucose backbone substituted by sulfate-ester groups, masked with ramifications, and containing other monosaccharide residues. We here report on the characterization of a novel glycoside hydrolase (FcnA) specific for the degradation of sulfated fucans. This glycoside hydrolase was purified to electrophoretic homogeneity from a Flavobacteriaceae referred to as SW5. The gene fcnA was cloned and sequenced (3021 nucleotides), and the protein (1007 amino acids) was produced in Escherichia coli. FcnA exhibited a modular architecture consisting of a 400-residue-long N-terminal domain followed by three repeated domains predicted to adopt an immunoglobulin fold and by an 80-amino acid-long C-terminal domain. A truncated recombinant protein encompassing the N-terminal domain and the immunoglobulin-like repeats was shown to retain the enzyme activity. The N-terminal catalytic domain shared 
25% of sequence identity with two patented fucanase genes, and these three fucanases delineate a new family of glycoside hydrolases. As shown by size-exclusion chromatography (SEC) and 1H-NMR analyses, the fucanase FcnA proceeds according to an endolytic mode of action and cleaves the -(1
4) glycosidic linkages within the blocks of repeating motifs [4)--L-fucopyranosyl-2,3-disulfate-(13)--L-fucopyranosyl-2-sulfate-(1]n.
Key words: brown algae / Flavobacteriaceae / fucanase / NMR / sulfated polysaccharide
The nucleotide sequence reported in this article has been submitted to the EMBL Nucleotide Sequence Database with accession number AJ877239.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  T. Barbeyron, S. L'Haridon, G. Michel, and M. Czjzek Mariniflexile fucanivorans sp. nov., a marine member of the Flavobacteriaceae that degrades sulphated fucans from brown algae Int J Syst Evol Microbiol, September 1, 2008; 58(9): 2107 - 2113. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Flament, T. Barbeyron, M. Jam, P. Potin, M. Czjzek, B. Kloareg, and G. Michel Alpha-Agarases Define a New Family of Glycoside Hydrolases, Distinct from Beta-Agarase Families Appl. Envir. Microbiol., July 15, 2007; 73(14): 4691 - 4694. [Abstract] [Full Text] [PDF]
|
|