Carbon Nitride Nanosheet-Supported Porphyrin: A New Biomimetic Catalyst for Highly Efficient Bioanalysis

A highly efficient biomimetic catalyst was fabricated based on ultrathin carbon nitride nanosheets (C3N4)-supported cobalt(II) proto-porphyrin IX (CoPPIX). The periodical pyridinic nitrogen units in C3N4 backbone could serve as electron donors for great affinity with Co2+ in PPIX, which resembled the local electronic structure as vitamin B12 and heme cofactor of hemoglobin. UV-vis kinetics and electrochemistry revealed its competitive (electro)catalysis with conventional peroxidase, while X-ray photoelectron spectroscopy and theoretical calculations suggest that the rehybridization of Co 3d with N orbitals from the backside can result in significant changes in enthalpy and charge density, which greatly promoted the activity of CoPPIX. The prepared nanocatalyst was further conjugated with streptavidin via multiple amines on the edge plane of C3N4 for facile tagging. Using biotinylated molecular beacon as the capture probe, a sensitive electrochemiluminescence-based DNA assay was developed via the electroreduction of H2O2 as the coreactant after the hairpin unfolded by the target, exhibiting linearity from 1.0 fM to 0.1 nM and a detection limit of 0.37 fM. Our results demonstrate a new paradigm to rationally design inexpensive and durable biomimics for electrochemiluminescence quenching strategy, showing great promise in bioanalytical applications.