Enhanced Stability of Enzyme Immobilized in Rationally Designed Amphiphilic Aerogel and Its Application for Sensitive Glucose Detection

Natural enzyme complex with the subunits cooperating with each other could catalyze cascade reactions in biological system but, just like the limitation of free-floating natural enzymes, usually suffer from deactivation in harsh environment such as high temperature. In this study, a purpose-driven design of amphiphilic aerogel working as the enzymes-immobilization substrate to form the multienzyme complex (MEC) was demonstrated. The aerogel was synthesized only by a single polymer poly­(vinyl alcohol) (PVA) as well as a surface modulator maleic acid (MA), the incorporation of which tunes the surface wettability. The usage of the amphiphilic aerogel may do favor for multienzyme immobilization, conserving the enzyme conformation as well as stabilizing the enzymes in high temperature. As a typical example, glucose oxidase and hemin were firmly coimmobilized in the aerogel matrix and actively catalyze the cascade reactions of (i) glucose to gluconic acid and (ii) 3,3,5,5-tetramethylbenzidine (TMB) to its oxidized state. The enzymes could resist the degradation under high temperature (70–100 °C) which is witnessed by the rate of decrease in activity was progressively slackened. Taking the advantage of the chromogenic reaction of TMB, a glucose sensor based on aerogel-enzyme composite for glucose detection in whole blood and sweat was established, exhibiting reliable results and satisfactory recovery. The modified aerogel could also withstand multiple physical deformation meantime maintaining good adsorption capacity as well as catalytic performance. The enzymes-loading aerogel model may hopefully contribute to composing sensors based on other analytes.