Transport limited adsorption experiments give a new lower estimate of the turnover frequency of Escherichia coli hydrogenase 1
Résumé
Protein Film Electrochemistry is a technique in which a redox enzyme is directly wired to an
electrode, which substitutes for the natural redox partner. In this technique, the electrical current
flowing through the electrode is proportional to the catalytic activity of the enzyme. However, in
most cases, the amount of enzyme molecules contributing to the current is unknown and the
absolute turnover frequency cannot be determined. Here, we observe the formation of
electrocatalytically active films of E. coli hydrogenase 1 by rotating an electrode in a
sub-nanomolar solution of enzyme. This process is slow, and we show that it is mass-transport
limited. Measuring the rate of the immobilization allows the determination of an estimation of the
turnover rate of the enzyme, which appears to be much greater than that deduced from solution
assays under the same conditions
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Revised version -- Transport limited adsorption shows E coli Hyd 1 is more active on an electrode than in solution.pdf (1.04 Mo)
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