Colorimetric Biosensor Based on Magnetic Enzyme and Gold Nanorods for Visual Detection of Fish Freshness

Histamine, an important safety index for aquatic products, can also be used as a freshness indicator for red-fleshed fish. In this work, magnetic graphene oxide (Fe3O4@GO, MGO) was applied to immobilize diamine oxidase (DAO) through a method of adsorption and covalent bonding. Under the optimized conditions, magnetic DAO prepared by adsorption immobilization had a higher enzyme activity than that of free enzyme, which was selected for the sensor construction. A colorimetric biosensor based on magnetic DAO induced etching of gold nanorods (AuNRs) was developed for the detection of histamine in fish. The developed biosensor showed an excellent response toward histamine with a low detection limit of 1.23 mu M and had negligible interference from other diamines. With increasing the histamine concentration, the AuNRs after the reaction exhibited colors ranging from dark green to blue-green, blue, purple, red, and colorless. The etching induced multicolor change of AuNRs indicated the presence of different contents of histamine in mackerel during storage, and was consistent with the overall change in the content of the total volatile basic nitrogen (TVB-N). Thus, it was indicated that the proposed colorimetric biosensor with a naked-eye-detectable readout has a great potential to evaluate the freshness of red-fleshed fish high in histamine.