Leiden University Scholarly Publications

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Nami, F. (2017)

Enzymatic reduction of oxygen by small laccase. A rapid freeze-quench EPR study

Doctoral Thesis
Since the discovery of enzymes as biological catalysts, the mechanism of enzymatic reactions has been a key question to enzymologists. Elucidating the reaction kinetics and the nature of enzymatic intermediates are necessary to understand such reactions. An important challenge in these studies is the limited lifetime of such intermediates, usually on the time scale of milliseconds to seconds. Therefore, a suited trapping method is required. Rapid freeze-quench (RFQ) is a proven technique to trap the intermediates on the time scale of milliseconds. The RFQ technique has been mostly combined with electron paramagnetic resonance (EPR) spectroscopy to reveal the nature of the
paramagnetic intermediates involved in enzymatic reactions. In this thesis, we improve and extend the combination of conventional RFQ technique with EPR spectroscopy up to a microwave frequency of 275 GHz to trap and characterize the intermediates involved in the enzymatic reduction of O2 by small laccase...Show more Since the discovery of enzymes as biological catalysts, the mechanism of enzymatic reactions has been a key question to enzymologists. Elucidating the reaction kinetics and the nature of enzymatic intermediates are necessary to understand such reactions. An important challenge in these studies is the limited lifetime of such intermediates, usually on the time scale of milliseconds to seconds. Therefore, a suited trapping method is required. Rapid freeze-quench (RFQ) is a proven technique to trap the intermediates on the time scale of milliseconds. The RFQ technique has been mostly combined with electron paramagnetic resonance (EPR) spectroscopy to reveal the nature of the
paramagnetic intermediates involved in enzymatic reactions. In this thesis, we improve and extend the combination of conventional RFQ technique with EPR spectroscopy up to a microwave frequency of 275 GHz to trap and characterize the intermediates involved in the enzymatic reduction of O2 by small laccase (SLAC).Show less
Rapid-freeze quench
Multi-frequency EPR
Enzyme mechanism
Small laccase
All authors
Nami, F.
Supervisor
Groenen, E.J.J.; Canters, G.W.
Committee
Benatti, M.; Hagen, W.R.; Tepper, A.W.J.W.; Eliel, E.R.; Orrit, M.A.G.J.
Qualification
Doctor (dr.)
Awarding Institution
Institute of Physics (LION), Science, Leiden University
Date
2017-03-07
Title of host publication
Casimir PhD Series
ISBN (print)
9789085932895

Publication Series

Name
2017-04

Funding

Sponsorship
NWO