Institute of Mass Spectrometry, School of Materials Science and Chemical Engineering, Ningbo University, Ningbo, Zhejiang 315211, China.
Kweichow Moutai Group, Renhuai, Guizhou 564501, China; Guizhou Key Laboratory of Microbial Resources Exploration in Fermentation industry, Kweichow Moutai Group, Zunyi 564501, China.
Int J Biol Macromol. 2024 May;268(Pt 1):131870. doi: 10.1016/j.ijbiomac.2024.131870. Epub 2024 Apr 24.
As one of the most important industrial enzymes, α-amylase is widely used in food processing, such as starch sugar and fermentation, bringing high added value to industry of more than a trillion dollars. We developed a multi-enzyme system (Glu&Gox@Cu-MOF-74) prepared by embedding α-glucosidase (Glu) and glucose oxidase (Gox) into the biomimetic metal-organic framework Cu-MOF-74 using in situ encapsulation within 15 min at room temperature for efficient and sensitive detection of α-amylase activity. Benefitting from the remarkable peroxidase-mimicking property and rigid skeleton of Cu-MOF-74, the biocatalytic platform exhibited excellent cascade activity and tolerance in various extremely harsh environments compared to natural enzymes. On this basis, a cascade biocatalytic platform was constructed for the detection of α-amylase activity with wide linear range (5-100 U/L) and low limit of detection (1.45 U/L). The colorimetric cascade scheme is important for the sensitive and selective determination of α-amylase in complex fermentation samples, and the detection time is short (∼0.5 h). This work provides new ideas for the detection of α-amylase based on the cascade amplification method.
作为最重要的工业酶之一，α-淀粉酶广泛应用于食品加工，如淀粉糖和发酵，为价值超过万亿美元的工业带来了高附加值。我们开发了一种多酶体系（Glu&Gox@Cu-MOF-74），通过在室温下 15 分钟内原位包埋，将α-葡萄糖苷酶（Glu）和葡萄糖氧化酶（Gox）嵌入仿生金属有机骨架 Cu-MOF-74 中，用于高效、灵敏地检测α-淀粉酶活性。得益于 Cu-MOF-74 的显著过氧化物酶模拟特性和刚性骨架，与天然酶相比，该生物催化平台在各种极端恶劣环境中表现出优异的级联活性和耐受性。在此基础上，构建了用于检测α-淀粉酶活性的级联生物催化平台，具有较宽的线性范围（5-100 U/L）和较低的检测限（1.45 U/L）。比色级联方案对于复杂发酵样品中α-淀粉酶的灵敏和选择性测定非常重要，检测时间短（约 0.5 小时）。这项工作为基于级联放大方法的α-淀粉酶检测提供了新的思路。
