title: Development of a high-throughput liquid state assay for lipase activity using natural substrates and rhodamine B.
abstract: A fluorescence-based assay for the determination of lipase activity using rhodamine B as an indicator, and natural substrates such as olive oil, is described. It is based on the use of a rhodamine B-natural substrate emulsion in liquid state, which is advantageous over agar plate assays. This high-throughput method is simple and rapid and can be automated, making it suitable for screening and metagenomics application. Reaction conditions such as pH and temperature can be varied and controlled. Using triolein or olive oil as a natural substrate allows monitoring of lipase activity in reaction conditions that are closer to those used in industrial settings. The described method is sensitive over a wide range of product concentrations and offers good reproducibility.
authors.affiliation: Centre de recherche sur les matériaux lignocellulosiques, Université du Québec à Trois-Rivières, Trois-Rivières, Québec G9A 5H7, Canada; PROTEO, Université Laval, Québec, Québec G1V 4G2, Canada.
authors.forename: Ximena
authors.initials: X
authors.lastname: Zottig
authors.affiliation: Centre de recherche sur les matériaux lignocellulosiques, Université du Québec à Trois-Rivières, Trois-Rivières, Québec G9A 5H7, Canada; PROTEO, Université Laval, Québec, Québec G1V 4G2, Canada.
authors.forename: Fatma
authors.initials: F
authors.lastname: Meddeb-Mouelhi
authors.affiliation: Centre de recherche sur les matériaux lignocellulosiques, Université du Québec à Trois-Rivières, Trois-Rivières, Québec G9A 5H7, Canada; PROTEO, Université Laval, Québec, Québec G1V 4G2, Canada. Electronic address: marc.beauregard@uqtr.ca.
authors.forename: Marc
authors.initials: M
authors.lastname: Beauregard
mesh_terms: D003124:Colorimetry; D057166:High-Throughput Screening Assays; D008049:Lipase / Q000378:metabolism*; D012235:Rhodamines / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D012235:Rhodamines; D008049:Lipase; C029773:rhodamine B
keywords: Fluorescence; Lipase activity; Olive oil; Rhodamine B