Immobilization of FeFe-hydrogenase on black TiO₂ nanotubes as biocathodes for the hydrogen evolution reaction

Hydrogenases are attractive biocatalysts for utilization in electrochemical devices as potential replacement for Pt in hydrogen evolving electrodes. In this work, we investigate the immobilization of ferredoxin tagged FeFe-hydrogenase (Fd-HydA1) on black TiO₂ nanotubes (bTNTs), with uniform nanotube opening diameters of 140 nm. By utilizing an immunogold labelling method, we show that the enzymes attach on the top surface of the bTNTs film rather than on the inner nanotube walls, reflecting the difficulty to insert enzymes into high aspect-ratio nanomaterials for O₂-shielding. Nevertheless, cyclic voltammetry demonstrates direct electron transfer between Fd-HydA1 and bTNTs for the hydrogen evolution reaction (HER) in neutral media. This work provides new insight towards design of new nanostructured electrodes for enzyme immobilization.