▎ 摘　　要

NOVELTY - Method for immobilizing glucose oxidase in oxidized graphite, involves dissolving oxidized graphene in distilled water, mixing oxidized graphene turbid liquid, adding e.g. 1-ethide-3-(3-dimethyl aminopropyl) carbodiimide into turbid liquid, centrifuging the mixture, drying the precipitate to obtain esterified oxidized graphene, uniformly dispersing activated concanavalin A in turbid liquid, centrifuging the mixture, washing the precipitate, adding glucose oxidase solution into precipitate, centrifuging the mixture, and freeze-drying the precipitate. USE - The method is useful for immobilizing glucose oxidase in oxidized graphite (claimed). ADVANTAGE - The method improves activity of immobilized enzyme to 150-195 U/mg, and reduces loss of enzyme activity. DETAILED DESCRIPTION - Method for immobilizing glucose oxidase in oxidized graphite, involves dissolving oxidized graphene in distilled water, mixing oxidized graphene turbid liquid until 1 mg/ml concentration is obtained by ultrasonically, adding 50 mu l oxidized graphene, 100 mu g N-hydroxy sulfo-succinimide and 44.1 mu g 1-ethide-3-(3-dimethyl aminopropyl) carbodiimide into the turbid liquid, using 0.1 M 2-(N-morpholino) ethanesulfonic acid buffer solution having pH of 6 to maintain constant volume of 1-ethide-3-(3-dimethyl aminopropyl) carbodiimide with final concentration of 20 mM, centrifuging the mixture for 30 minutes at high speed, discarding the supernatant, washing the precipitate using 0.05 M 2-(N-morpholino) ethanesulfonic acid buffer solution, drying the precipitate in vacuum for 24 hours to obtain esterified oxidized graphene, adding activated concanavalin A solution having pH of 7 into 1 mg/ml esterified oxidized graphene, uniformly dispersing activated concanavalin A solution in turbid liquid, centrifuging the mixture, mixing 50 mu l esterified oxidized graphene with 300-1500 mu l (0.1 mg/ml) concanavalin A solution, centrifuging the mixture at 4 degrees C for 60 minutes, discarding the supernatant, washing the precipitate using 0.05 M 2-(N-morpholino) ethanesulfonic acid buffer solution, adding 0.5 mg/ml glucose oxidase solution having pH of 4.5-8 into precipitate, where the mass ratio of concanavalin A, and glucose oxidase is 1:5, centrifuging the mixture, discarding the supernatant, washing the precipitate using phosphate buffer solution (PBS), and freeze-drying the precipitate under vacuum for 24 hours to obtain immobilized glucose oxidase.