Nano-Biocatalysts of Cyt c@ZIF-8/GO Composites with High Recyclability via a de Novo Approach

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.