Fabrication and application of a novel electrochemical biosensor based on a mesoporous carbon sphere@UiO-66-NH2/Lac complex enzyme for tetracycline detection

Abstract

The overuse of tetracycline results in a threat to human, poultry and livestock health. An enzymatic electrochemical biosensor is an ideal alternative method for accurate and rapid tetracycline detection, while the unstable and easily deactivated nature of the enzyme limits its development. To overcome these limitations, a highly sensitive enzymatic electrochemical biosensor for the determination of tetracycline is developed in this work based on a complex enzyme which was constructed using a mesoporous carbon sphere@UiO-66-NH₂ (MCS@UiO-66-NH₂) core–shell composite with embedded laccase (Lac). Compared to pure MCS and UiO-66-NH₂, the MCS@UiO-66-NH₂ core–shell composite has an advantageous mesoporous structure (pore diameter >8 nm), which is suitable for the immobilization of small laccase. The biosensor based on the complex enzyme exhibits a superior activity and enhanced stability as compared with that made using a pure enzyme because the mesoporous structure of the MCS@UiO-66-NH₂ composite can effectively protect the laccase against inactivation and denaturation. Besides, its high specific surface area and good conductivity are beneficial to enzyme immobilization and electron transfer in the modified electrode. The biosensor based on this complex enzyme exhibits a relatively low detection limit of 8.94 × 10⁻⁷ mol L⁻¹ and a detection range of 1.0 × 10⁻⁶–6.0 × 10⁻⁵ mol L⁻¹ for tetracycline detection. Furthermore, the developed biosensor possesses good long-term stability, selectivity and reproducibility, indicating its potential application for tetracycline determination in actual food. This research work provides a prospective solution to resolve the stability and inactivation problems of enzymatic electrochemical biosensors in different application scenarios.