Covalent organic frameworks (COFs), as one of the most significant members of the porous organic frameworks, have been well used in the photocatalysis owing to their outspread π-conjugated framework, high crystallinity and regular pore structure. Herein, after reducing the labile imine-linked COF-300 to the more stable amine-linked COF-300-AR, we for the first time demonstrated that COF-300-AR was the light-responsive oxidase mimic. COF-300-AR exhibited excellent oxidase-mimicking activity under purple light stimulation (λ = 400 nm), which can catalyze the oxidation of classical substrates such as 3,3',5,5'-tetramethylbenzydine (TMB) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) by the formation of •OH and O2•- free radicals in the presence of dissolved oxygen. The COF-300-AR oxidase mimic has outstanding advantages of easy light control, high stability, good reusability, and highly catalytic oxidation capacity and has been applied to detect glutathione (GSH) levels in HL60 cells with good selectivity and high sensitivity. This study will broaden the sensing applications of COFs and offer a promising build block for the construction of artificial enzymes.