Author:

Keywords:

analytical methods, enzymes, fluorescence, microscopy, kinetics, single-molecule studies, conformational dynamics, candida-antarctica, lipase-b, molecule, spectroscopy, f-1-atpase, Science & Technology, Physical Sciences, Chemistry, Multidisciplinary, Chemistry, CONFORMATIONAL DYNAMICS, CANDIDA-ANTARCTICA, LIPASE-B, MOLECULE, SPECTROSCOPY, F-1-ATPASE, Candida, Catalysis, Hydrolysis, Kinetics, Lipase, 03 Chemical Sciences, Organic Chemistry, 34 Chemical sciences

Abstract:

Real-time measurement of the catalysis and substrate kinetics of a single-enzyme hydrolysis reaction is demonstrated with confocal fluorescence microscopy (CFM; see picture, green = CFM beam). A single lipase is shown to have a broad range of conformations; each conformation contributes to the overall enzymatic activity, an observation that is often masked by ensemble measurements.