This research was supported by the SISITOMAS European network and an EC Marie Curie fellowship (K.V.). The FWO-vlaanderen, the federal science policy through IAP/V/03 and the KULeuven through GOA 01/2 are thanked. O.F. thanks fruitful discussions with Attila Szabo and Irina Gopich. D.L. thanks the IWT for financial support. CALB=Lipase B from Candida antarctica.
Single-Enzyme Kinetics of CALB-Catalyzed Hydrolysis†
Article first published online: 23 DEC 2004
Copyright © 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Angewandte Chemie International Edition
Volume 44, Issue 4, pages 560–564, January 14, 2005
How to Cite
Velonia, K., Flomenbom, O., Loos, D., Masuo, S., Cotlet, M., Engelborghs, Y., Hofkens, J., Rowan, A. E., Klafter, J., Nolte, R. J. M. and de Schryver, F. C. (2005), Single-Enzyme Kinetics of CALB-Catalyzed Hydrolysis. Angew. Chem. Int. Ed., 44: 560–564. doi: 10.1002/anie.200460625
- Issue published online: 11 JAN 2005
- Article first published online: 23 DEC 2004
- Manuscript Revised: 7 JUL 2004
- Manuscript Received: 11 MAY 2004
- analytical methods;
- fluorescence microscopy;
- single-molecule studies
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.