Abstract: his study demonstrated for the first time that bromelin exhibited peroxidase-like activity. Enzymatic characterization confirmed that the optimal temperature and pH for its peroxidase-like activity were respectively 40 ℃ and 5.0, and with H2O2and TMB as substrates, the specific enzyme activities were 16.58 U/mg and 18.41 U/mg, respectively. Furthermore, a cascade catalytic system was constructed by coupling bromelin with glucose oxidase, enabling broad-range and highly sensitive glucose detection with a linear range of 5－400 μmol/L and a detection limit of 1.5 μmol/L. Additionally, the system demonstrated excellent selectivity and resistance to interference for glucose detection, enabling its effective application in the analysis of human blood and urine samples. Therefore, this research not only provided an efficient, sensitive, and visual detection method for diabetes diagnosis, characterized by operational simplicity and excellent cost-effectiveness, but also successfully expanded the application scope of bromelain in enzymology, bringing new vitality into enzymatic research and applications.

Key words: cascade catalysis, sensitive detection, selectivity, anti-interference ability