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.