title: The cell-wall active mannuronan C5-epimerases in the model brown alga Ectocarpus: From gene context to recombinant protein.
abstract: Mannuronan C5-epimerases (ManC5-Es) catalyze in brown algae the remodeling of alginate, a major cell-wall component which is involved in many biological functions in these organisms. ManC5-Es are present as large multigenic families in brown algae, likely indicating functional specificities and specializations. ManC5-Es control the distribution pattern of (1-4) linked β-d-mannuronic acid (M) and α-l-guluronic acid (G) residues in alginates, giving rise to widely different polysaccharide compositions and sequences, depending on tissue, season, age, or algal species. As such they are also a source of powerful new tools for the biotechnological and enzymatic processing of alginates, to match the growing interest for food hydrocolloids and in biomedical and nanotechnological applications. We report here the first heterologous production of a ManC5-E of brown algal origin that is successfully refolded in an active form. The activity was measured by1H NMR and by an indirect enzymatic assay using a known bacterial alginate lyase. The transcript expression as a function of the developmental program of the brown alga Ectocarpus, together with the bioinformatic analyses of the corresponding gene context of this multigenic family, is also presented.
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France.
authors.forename: Richard
authors.initials: R
authors.lastname: Fischl
authors.affiliation: DuPont Nutrition Biosciences ApS, Aarhus, Denmark.
authors.forename: Kresten
authors.initials: K
authors.lastname: Bertelsen
authors.affiliation: FR2424, Station Biologique de Roscoff, CS 90074, Roscoff, France.
authors.forename: Fanny
authors.initials: F
authors.lastname: Gaillard
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France.
authors.forename: Susana
authors.initials: S
authors.lastname: Coelho
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France.
authors.forename: Gurvan
authors.initials: G
authors.lastname: Michel
authors.affiliation: DuPont Nutrition Biosciences ApS, Aarhus, Denmark.
authors.forename: Markus
authors.initials: M
authors.lastname: Klinger
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Boyen
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France cecile.herve@sb-roscoff.fr mirjam.czjzek@sb-roscoff.fr.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Czjzek
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff, CS 90074, Roscoff, France cecile.herve@sb-roscoff.fr mirjam.czjzek@sb-roscoff.fr.
authors.forename: Cécile
authors.initials: C
authors.lastname: Hervé
mesh_terms: D000464:Alginates / Q000378:metabolism; D000595:Amino Acid Sequence; D002238:Carbohydrate Epimerases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002473:Cell Wall / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D000459:Phaeophyceae / Q000201:enzymology* / Q000235:genetics; D011134:Polysaccharides / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D011134:Polysaccharides; D011994:Recombinant Proteins; C007896:guluronic acid; D020723:Glucuronic Acid; C008324:mannuronic acid; D002238:Carbohydrate Epimerases; C086519:mannuronan c-5-epimerase
keywords: Ectocarpus; alginate; brown algae; mannuronan C5-epimerase