title: Droplet-based microfluidics platform for ultra-high-throughput bioprospecting of cellulolytic microorganisms.
abstract: Discovery of microorganisms producing enzymes that can efficiently hydrolyze cellulosic biomass is of great importance for biofuel production. To date, however, only a miniscule fraction of natural biodiversity has been tested because of the relatively low throughput of screening systems and their limitation to screening only culturable microorganisms. Here, we describe an ultra-high-throughput droplet-based microfluidic system that allowed the screening of over 100,000 cells in less than 20 min. Uncultured bacteria from a wheat stubble field were screened directly by compartmentalization of single bacteria in 20 pl droplets containing a fluorogenic cellobiohydrolase substrate. Sorting of droplets based on cellobiohydrolase activity resulted in a bacterial population with 17- and 7-fold higher cellobiohydrolase and endogluconase activity, respectively, and very different taxonomic diversity than when selected for growth on medium containing starch and carboxymethylcellulose as carbon source.
authors.affiliation: Institut de Science et d'Ingénierie Supramoléculaires (ISIS), Université de Strasbourg, CNRS UMR 7006, 8 allée Gaspard Monge, 67083 Strasbourg Cedex, France; Division Biotechnologies, Ets. J. Soufflet, quai Sarrail, 10400 Nogent-sur-Seine, France.
authors.forename: Majdi
authors.initials: M
authors.lastname: Najah
authors.affiliation: Institut de Science et d'Ingénierie Supramoléculaires (ISIS), Université de Strasbourg, CNRS UMR 7006, 8 allée Gaspard Monge, 67083 Strasbourg Cedex, France.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Calbrix
authors.affiliation: Institut de Science et d'Ingénierie Supramoléculaires (ISIS), Université de Strasbourg, CNRS UMR 7006, 8 allée Gaspard Monge, 67083 Strasbourg Cedex, France; Division Biotechnologies, Ets. J. Soufflet, quai Sarrail, 10400 Nogent-sur-Seine, France.
authors.forename: I Putu
authors.initials: IP
authors.lastname: Mahendra-Wijaya
authors.affiliation: École Supérieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI ParisTech), CNRS UMR 8231, 10 rue Vauquelin, 75231 Paris Cedex 05, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Beneyton
authors.affiliation: Institut de Science et d'Ingénierie Supramoléculaires (ISIS), Université de Strasbourg, CNRS UMR 7006, 8 allée Gaspard Monge, 67083 Strasbourg Cedex, France; École Supérieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI ParisTech), CNRS UMR 8231, 10 rue Vauquelin, 75231 Paris Cedex 05, France. Electronic address: andrew.griffiths@espci.fr.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Griffiths
authors.affiliation: Institut de Science et d'Ingénierie Supramoléculaires (ISIS), Université de Strasbourg, CNRS UMR 7006, 8 allée Gaspard Monge, 67083 Strasbourg Cedex, France; Division Biotechnologies, Ets. J. Soufflet, quai Sarrail, 10400 Nogent-sur-Seine, France. Electronic address: adrevelle@soufflet-group.com.
authors.forename: Antoine
authors.initials: A
authors.lastname: Drevelle
mesh_terms: D001419:Bacteria / Q000201:enzymology / Q000302:isolation & purification* / Q000378:metabolism*; D056804:Biofuels / Q000382:microbiology; D018533:Biomass; D065132:Bioprospecting / Q000379:methods*; D002482:Cellulose / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006868:Hydrolysis; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D012988:Soil Microbiology
chemical_list: D056804:Biofuels; D002482:Cellulose; D043366:Cellulose 1,4-beta-Cellobiosidase