title: Functional Metagenomics: Construction and High-Throughput Screening of Fosmid Libraries for Discovery of Novel Carbohydrate-Active Enzymes.
abstract: Activity-based metagenomics is one of the most efficient approaches to boost the discovery of novel biocatalysts from the huge reservoir of uncultivated bacteria. In this chapter, we describe a highly generic procedure of metagenomic library construction and high-throughput screening for carbohydrate-active enzymes. Applicable to any bacterial ecosystem, it enables the swift identification of functional enzymes that are highly efficient, alone or acting in synergy, to break down polysaccharides and oligosaccharides.
authors.affiliation: INSA, UPS, INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France.|UMR792 Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400, Toulouse, France.|UMR5504, CNRS, 31400, Toulouse, France.
authors.forename: Lisa
authors.initials: L
authors.lastname: Ufarté
authors.affiliation: INSA, UPS, INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France.|UMR792 Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400, Toulouse, France.|UMR5504, CNRS, 31400, Toulouse, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bozonnet
authors.affiliation: INSA, UPS, INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France.|UMR792 Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400, Toulouse, France.|UMR5504, CNRS, 31400, Toulouse, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Laville
authors.affiliation: INSA, UPS, INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France.|UMR792 Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400, Toulouse, France.|UMR5504, CNRS, 31400, Toulouse, France.
authors.forename: Davide A
authors.initials: DA
authors.lastname: Cecchini
authors.affiliation: INSA, UPS, INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France.|UMR792 Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400, Toulouse, France.|UMR5504, CNRS, 31400, Toulouse, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Pizzut-Serin
authors.affiliation: Laboratoire Ampère, CNRS UMR5005, Ecole Centrale de Lyon, Université de Lyon, 69134, Ecully, France.
authors.forename: Samuel
authors.initials: S
authors.lastname: Jacquiod
authors.affiliation: Laboratoire Ampère, CNRS UMR5005, Ecole Centrale de Lyon, Université de Lyon, 69134, Ecully, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Demanèche
authors.affiliation: Laboratoire Ampère, CNRS UMR5005, Ecole Centrale de Lyon, Université de Lyon, 69134, Ecully, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Simonet
authors.affiliation: Laboratoire Ampère, CNRS UMR5005, Ecole Centrale de Lyon, Université de Lyon, 69134, Ecully, France.
authors.forename: Laure
authors.initials: L
authors.lastname: Franqueville
authors.affiliation: INSA, UPS, INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France. veronese@insa-toulouse.fr.|UMR792 Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400, Toulouse, France. veronese@insa-toulouse.fr.|UMR5504, CNRS, 31400, Toulouse, France. veronese@insa-toulouse.fr.
authors.forename: Gabrielle Potocki
authors.initials: GP
authors.lastname: Veronese
mesh_terms: D001419:Bacteria / Q000201:enzymology* / Q000235:genetics; D002241:Carbohydrates / Q000235:genetics; D020536:Enzyme Activators / Q000378:metabolism; D004798:Enzymes / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D056186:Metagenomics / Q000379:methods*; D010957:Plasmids
chemical_list: D002241:Carbohydrates; D020536:Enzyme Activators; D004798:Enzymes
keywords: Carbohydrate-active enzymes; Complex glycans; Fosmidic libraries; High-throughput screening; Metagenomic DNA