(Invited) ATP-Independent Electroenzymatic Ammonia Production Using an Organic Redox Polymer-Immobilized Enzymatic System
Authors: Lee Yoo Seok , Minteer Shelley D. 
Published: July 2021

Consuming approximately 1% of the total fossil fuel of the world, the ammonia (NH3), a usable form to living organisms is produced by dinitrogen (N2) fixation. Currently, approximately 66% of NH3 is produced by the Haber–Bosch process using an iron catalyst with N2 and H2 at high temperatures and high pressures (400 °C and 20 MPa), which are estimated to be responsible for approximately 3% of CO2 emissions.1,2 As an alternative strategy, bioelectrosynthetic technique is studied for eco-friendly ammonia production using the nitrogenase, the only enzyme able to reduce N2 to NH3 at room temperature, neutral pH and ambient pressure. For the practical use of nitrogenase bioelectrocatalysis technology, it is indispensable to minimize the large input of chemical energy for the reduction of MoFe protein component of nitrogenase by hydrolysis of two ATP for each electron transferred (16 ATP per N2).3-5

Therefore, we have concentrated on approaches to bypass the reducing- and ATP-hydrolyzing properties of the dinitrogenase reductase by immobilizing the catalytic protein of nitrogenase, MoFe protein, on the electrode surface. One way to directly reduce the MoFe protein is a use of a redox polymer having thermodynamically strong reducing capability. Redox polymer immobilized bioelectrocatalysis facilitates an efficient electron transfer through self-exchange based conduction due to a high effective concentration of catalysts at the electrode surface and more efficient mediation. However, none of the previously reported redox polymers were capable of immobilized bioelectrocatalysis using nitrogenase.6-9

Thus, we report the MoFe nitrogenase immobilized at an electrode surface with a neutral red redox polymer which is used to reduce the MoFe protein (as mediator able to transfer the electrons to the MoFe independent of the Fe protein and of ATP hydrolysis) and support the mediated bioelectrocatalysis of N3
-, NO2
- and N2 to NH3 catalyzed by the MoFe protein. Representative Bulk bioelectrosynthetic experiments produced 209 ± 30 nmol NH3 nmol MoFe-1 h-1 from N2 reduction. 15N2 labeling experiments and NMR analysis were performed to confirm biosynthetic N2 reduction to NH3
.

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