A novel biosensor based on multienzyme microcapsules constructed from covalent-organic framework.

Electrochemical biosensors based on enzymes modified electrode are attracting special attention due to their broad applications. However, the immobilization of enzymes on electrode is always an important challenge because it's not conducive to conformational expansion of enzymes and affects the bioactivity of enzymes accordingly. Although the imobilization of enzymes in micropores of crystalline covalent-organic framework (COF) and metal-organic framework (MOF) to construct electrochemical biosensors based on pore embedding can achive good reuslts, their micropores can still not guarantee that the enzyme's conformation is well extended. Herein, a multienzyme microcapsules (enzymes@COF) containing glucose oxidase, horseradish peroxidase and acetylcholinesterase with a 600 nm-sized cavity and a shell of COF was used to construct electrochemical biosensors. The 600 nm-sized cavity ensures free conformation expansion of encapsulated enzymes and the shell of COF with good chemical stablity protects encapsulated enzymes against external harsh environments. And the specific catalytic substrates of the enzymes can infiltrated into the microcapsule through the pores of COF shell. So, the biosensor based on enzymes@COF microcapsules demonstrated preeminent performances as compared with those of enzymes assembled on electrode. The detection limits were 0.85 μM, 2.81 nM, 3.0×10 g/L, and the detection range were 2.83 μM-8.0 mM, 9.53 nM-7.0 μM, 10 g/L-10 g/L for glucose, H2O2 and malathion detection. This work shows that it is feasible to fabricate electrochemical sensors using enzymes@COF microcapsules.