A novel nanocomposite based on ordered graphitized mesoporous carbon (GMC) and amino acid ionic liquids (AAIL) is obtained through controlled surface modification of GMC with hydrophilic AAILs (1-ethyl-3-methylimidazolium alanine, EMIM[Ala]), which is used as a platform for a tyrosinase biosensor to detect phenol. The GMC–AAIL nanocomposite possesses a better biocompatibility and improved aqueous-phase dispersion than hydrophobic GMC alone, owing to the introduction of hydrophilic and biocompatible AAILs. Comparative studies revealed that the catalytic activity of tyrosinase for phenol in phosphate buffer solution (PBS) containing EMIM[Ala] was about ten times higher than that in pure PBS. By entrapping tyrosinase molecules into the mesopores of GMC, making use of the synergy effect of GMC and AAIL (the “interspace confinement effect”, the anti-fouling ability, and the biocompatible microenvironment), the GMC–AAIL-based biosensors display superior analytical performance to GMC-based ones in terms of signal-to-noise ratio, stability, repeatability, and working life. After 21-day storage, the electrode retained more than 90% of its initial response, indicating that surface modification of GMC with hydrophilic and biocompatible AAILs could significantly prolong the life of tyrosinase in vitro. The GMC10–EMIM[Ala]-based biosensor demonstrates a linear response for phenol concentrations from 0.1 to 10 µmol L−1 with a low detection limit of 20 nmol L−1 and sensitivity of 1385 mA cm−2 M−1. The GMC–AAIL nanocomposite proves to be a promising platform for enzyme-based biosensors and biocatalysis.
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