Platinum nanoparticle-deposited multi-walled carbon nanotubes as a NADH oxidase mimic: characterization and applications

The effective regeneration of bioactive NAD+ plays an important role in numerous dehydrogenase-dependent applications including biocatalysis and biosensing. However, this process usually suffers from high thermodynamic barrier, instability and high cost associated with natural enzymes. The emergence of nanomaterials with enzyme mimic characteristics has offered a potential alternative to many enzyme-catalyzed processes. Platinum nanoparticles (PtNPs), for example, have been extensively studied for their peroxidase- and oxidase-like activities. However, their behavior as a NADH oxidase mimic has barely been characterized in detail. Herein, we report a facile approach for preparing PtNP-deposited multi-walled carbon nanotubes (PtNPs@MWCNTs) as the nanozyme for NADH oxidation. Its enzymatic activity was investigated in depth, revealing that it is a NADH oxidase instead of a peroxidase and the catalytic process generates O2˙-, rather than OH˙ or 1O2, from dissolved O2. The recovery yield of bioactive NAD+ regeneration by the nanozyme could reach ∼100% with a total turnover number of ∼6000. Besides, it exhibited terrific electrochemical performance for NADH oxidation and sensing by greatly boosting the response and lowering the oxidation overpotential. It could also work on biomimetic cofactors with even higher activity. Finally, xylose dehydrogenase was immobilized with the nanozyme to constitute a hybrid bioelectrode for xylose sensing. The biosensor had a xylose detecting range of 5-400 μM with the limit of detection as low as 1 μM and can retain its performance after being reused several times. Our results suggest that the PtNPs@MWCNTs characterized as a NADH oxidase nanozyme hold great promise in the applications of biocatalysis and biosensing, which intensively deal with dehydrogenases and natural or biomimetic cofactors.