Nitrogenase is the
only biological catalyst that is known to be
able to convert nitrogen gas to ammonia. In microorganisms, the MoFe
catalytic protein of nitrogenase is reduced by a transient Fe protein
binding and separate hydrolysis of ATP. However, the requirement of
16 ATP molecules by the Fe protein for the 8 electron transfer is
an energy-intense caveat to the enzymatic synthesis of NH3 and is challenging from an electrochemical perspective. Thus, we
report the redox polymer-based ATP-free mediated electron-transfer
system of MoFe nitrogenase using cobaltocene-functionalized poly(allylamine)
(Cc-PAA), which is able to reduce the MoFe nitrogenase directly with
a low redox potential of −0.58 V vs SHE. An efficient immobilization
of MoFe nitrogenase via Cc-PAA allowed for the bioelectrocatalytic
reduction of N3
–, NO2
–, and N2 to NH3. Bulk bioelectrosynthetic
experiments produced 7 ± 2 and 30 ± 5 nmol of NH3 from NO2
– and N3
– reduction for 30 min, respectively. In addition, biosynthetic N2 reduction to NH3 was confirmed by 15N2 labeling experiments with NMR analysis. This mediated
electron-transfer approach of the immobilized nitrogenase using the
Cc-PAA redox polymer provides a valuable technological basis for scale-up
and industrial uses in the future of bioelectrosynthesis.