Rapid and ultrasensitive activity detection of α-amylase based on γ-cyclodextrin crosslinked metal-organic framework nanozyme.

α-Amylase plays a significant part in fermentation and the food industry, as this enzyme effectively regulates the content of different sugars in brewing systems and affects the yield and quality of alcoholic beverages. Nevertheless, current strategies suffer from unsatisfactory sensitivity and are time-consuming or are indirect methods which demand the assistance of tool enzymes or inhibitors. Therefore, they are unsuitable for the low bioactivity and non-invasive detection of α-amylase in fermentation samples. Rapid, sensitive, facile, and direct detection method of this protein remains challenging in actual applications. In this work, a nanozyme-based α-amylase assay was constructed. The colorimetric assay used the interaction between α-amylase and γ-cyclodextrin (γ-CD) which crosslinks MOF-919-NH. The determination mechanism bases on the hydrolysis of γ-CD by α-amylase, resulting in increased peroxidase-like bioactivity of the released MOF nanozyme. The detection limit was 0.12 U L with a wide linear range (0-200 U L) and excellent selectivity. Additionally, the proposed detection method was successfully utilized in distilled yeasts to verify analytical capability in fermentation samples. The exploration of this nanozyme-based assay not only provides a convenient and effective strategy for enzyme activity determination in food industry, but also has promotion significance in clinical diagnosis and pharmaceutical production.