Colorimetric detection of diamine using diamine oxidase and horseradish peroxidase co-incorporated hybrid microsphere as biomimetic cascade enzymes.

Dual-enzyme co-embedded materials have shown high potential for achieving efficient detection due to the convenience of two-enzyme cascade reactions. Herein, we developed a dual-enzyme hybrid microsphere (HM) based biosensor to detect diamines (histamine was included for ease of description) in aquatic products. The HM was made from diamine oxidase, horseradish peroxidase, and copper phosphate through the biomineralization method. Under optimal conditions, the system displayed linear color response to histamine of different concentrations ranging from 0 to 200 μg/mL. The detection limit of histamine was 0.15 μg/mL, showing higher sensitivity than the two-step free enzyme assay. Moreover, the detection system exhibited good specificity to diamines. The method was used to detect diamines in commercial samples, and the results were compared with those measured by the high-performance liquid chromatography method. Overall, the proposed assay exhibited high potential in diamine quantification and was readily extended to other cascade enzymatic reaction-based detection strategies.