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Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
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March 18, 2012
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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.
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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
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Romero, E., Robinson, R., Sobrado, P. Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry. J. Vis. Exp. (61), e3803, doi:10.3791/3803 (2012).
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