Presteady state kinetic analysis of riboflavin synthase |
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Authors: | Illarionov Boris Haase Ilka Bacher Adelbert Fischer Markus Schramek Nicholas |
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Affiliation: | Lehrstuhl für Organische Chemie und Biochemie, Technische Universit?t Munich, Lichtenbergstrasse 4, D-85747 Garching, Germany. |
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Abstract: | Riboflavin synthase catalyzes a mechanistically complex dismutation affording riboflavin and 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione from 6,7-dimethyl-8-ribityllumazine. The kinetics of the enzyme from Escherichia coli were studied under single turnover conditions. Stopped flow as well as quenched flow experiments documented the transient formation of a pentacyclic reaction intermediate. No other transient species were sufficiently populated to allow detection. The data are best described by a sequence of one second order and one first order reaction. |
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