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Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
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Chapters
- 00:05Title
- 01:37Anaerobic Buffer
- 02:24Removing Oxygen from the Stopped-flow System
- 03:22Key Solutions
- 04:45Reductive Half-reaction: Monitoring Flavin Reduction
- 07:31Oxidative Half-reaction: Monitoring Flavin Oxidation
- 10:10Stopped Flow: Spectral Forms of Af SidA
- 11:27Conclusion
We describe the use of a stopped-flow instrument to investigate both the reductive and oxidative half-reactions of Aspergillus fumigatus siderophore A (SidA), a flavin-dependent monooxygenase. We then show the spectra corresponding to the species in the reaction of SidA and we calculate the rate constants for their formation.
Tags
Stopped-flow SpectrophotometryFlavin-dependent MonooxygenaseAspergillus Fumigatus Siderophore A (SidA)Hydroxylation Of OrnithineBiosynthesis Of Hydroxamate SiderophoresReductive Half-reactionOxidative Half-reactionFADNADPHC4a-hydroperoxyflavin IntermediateMeasurement Of RatesSpectral FormsStopped-flow InstrumentAnaerobic Glove BoxSingle ModeDouble Mixing Settings
