Conformational rigidification of glucose oxidase induced by Ti3C2 MXene: A spectroscopic and computational study on non-covalent allosteric-like activation.

Glucose oxidase (GOx) is crucial for biocatalysis and cancer starvation therapy, yet its catalytic efficiency is constrained by inherent conformational flexibility. Herein, we report a non-covalent allosteric-like activation strategy for GOx using Ti3C2 MXene nanosheets as a conformational modulator. By integrating multispectral analyses, enzyme kinetics, and molecular dynamics simulations, we demonstrate that MXene spontaneously binds GOx via hydrogen bonding and van der Waals forces, forming a stable ground-state complex. This binding induces pronounced conformational rigidification (notably increased α-helix content), which directly translates into enhanced catalysis-lowered Michaelis constant and elevated maximum velocity, reflecting both improved substrate affinity and accelerated catalytic turnover. Molecular docking, alongside molecular dynamic simulations, reveals that the MXene binding site is spatially decoupled from the catalytic active center, with a persistent hydrogen-bonding network rigidifying the enzyme scaffold and strongly suggesting an allosteric-like regulatory mechanism. In vitro, the MXene-GOx composite exerts significantly enhanced, glucose-dependent cytotoxicity against HeLa cancer cells via amplified starvation and oxidative stress, directly linking the enzymatic enhancement to therapeutic efficacy. This work elucidates a fundamental mechanism of nanomaterial-induced enzyme activation and offers a rational framework for designing high-performance GOx-based systems through non-covalent conformational modulation.