We demonstrate electrochemical control of the nitrogenase MoFe protein, in the absence of Fe protein or ATP, using europium(iii/ii) polyaminocarboxylate complexes as electron transfer mediators. This allows the potential dependence of proton reduction and inhibitor (CO) binding to the active site FeMo-cofactor to be established. Reduction of protons to H2 is catalyzed by the wild type MoFe protein and β-98Tyr→His and β-99Phe→His variants of the MoFe protein at potentials more negative than -800 mV (vs. SHE), with greater electrocatalytic proton reduction rates observed for the variants compared to the wild type protein. Electrocatalytic proton reduction is strongly attenuated by carbon monoxide (CO), and the potential-dependence of CO binding to the FeMo-cofactor is determined by in situ infrared (IR) spectroelectrochemistry. The vibrational wavenumbers for CO coordinated to the FeMo-cofactor are consistent with earlier IR studies on the MoFe protein with Fe protein/ATP as reductant showing that electrochemically generated states of the protein are closely related to states generated with the native Fe protein as electron donor.
Bibliographical noteFunding Information:
This work was supported financially by the European Research Council (ERC, EnergyBioCatalysis-ERC-2010-StG-258600, K. A. V. and P. A. A.) and Engineering and Physical Sciences Research Council (EPSRC) Small Equipment Award EP/K031503/1 (K. A. V.). P. P. was supported by a scholarship from the Thai government. We are grateful to Henry Waite for carrying out initial Eu-L experiments and to Ricardo Hidalgo for the electrochemical flow cell design. This material is based upon work supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences (DE-SC0010687 and DE-SC0010834 to LCS and DRD).
© The Royal Society of Chemistry.