To improve the stability and recyclability of enzymes immobilized on metal-organic frameworks (MOFs), graphene oxide (GO) with surface oxygen-rich functional groups was selected to form ZIF-8/GO nanocomposites with the zeolitic imidazolate framework (ZIF-8) for cytochrome c (Cyt c) immobilization. It was found that the functional groups on the GO surface were involved in the growth of ZIF-8 without affecting the crystal structure but their particle size was reduced to about 200 nm. The storage stability and resistance to organic solvents of Cyt c were obviously improved after the immobilization on the ZIF-8/GO nanocomposite. On one hand, compared with Cyt c@ZIF-8 and Cyt c@GO with 30 and 60% protein leakage, Cyt c@ZIF-8/GO displayed little protein leakage after 60 h of storage. On the other hand, Cyt c@ZIF-8/GO retained a residual activity of approximately 100% after being stored in ethanol and acetone for 2 h, whereas the free enzyme, Cyt c@ZIF-8, and Cyt c@GO retained only about 10, 50, and 40%, respectively. In addition, the Cyt c@ZIF-8/GO nanocomposites can be utilized up to four cycles with virtually no loss of activity and may be further applied on H2O2 biosensing systems. The synergistic effect between MOFs and GO in ZIF-8/GO nanocomposites provides infinite possibilities as immobilized enzyme carriers.
Keywords: cytochrome c; enzyme immobilization; graphene oxide; metal-organic frameworks; zeolitic imidazolate framework.