Hierarchical mesoporous ZIF-8–immobilized enzyme microreactor coupled with HPLC for screening of α-glucosidase inhibitors from green tea

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2025-05-06 DOI:10.1007/s00604-025-07201-1

Ruobing Wu, Xiangrong Wu, Chenjia Jiang, Haodong Zhou, Jinhua Zhu, Zaved H. Khan

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Abstract

Hierarchical mesoporous ZIF-8 (HZIF-8) was synthesized using a thermosensitive hydrogel as a soft template. Zinc ions (Zn²⁺) were uniformly dispersed on the hydrogel surface through metal-ion coordination with salicylic acid and melamine, followed by coordination with 2-methylimidazole at room temperature. After the thermal removal of salicylic acid and melamine, HZIF-8 was obtained which exhibited a bimodal pore structure comprising both micropores and mesopores. α-Glucosidase (α-Glu) was encapsulated into HZIF-8 via an in-situ diffusion method to construct an immobilized α-Glu microreactor. The morphological and structural characteristics of HZIF-8 before and after α-Glu immobilization were thoroughly characterized using Fourier transform infrared spectroscopy (FT-IR), electron microscopy, X-ray diffraction (XRD), and Brunauer-Emmet-Teller (BET) analysis. The immobilization optimization studies revealed that the optimal immobilization time was 3 h, and the ideal mass ratio of HZIF-8 to α-Glu was 5:2 (w/w). Under these conditions, the α-Glu loading capacity reached 82.09 μg/mg, and the immobilized α-Glu microreactor still retained 70% enzyme activity after seven cycles. The application of the HZIF-8-α-Glu microreactor was verified by coupling with high performance liquid chromatography (HPLC) for the offline screening of α-Glu inhibitors in green tea extract. The experimental results demonstrated the application of the immobilized α-Glu microreactor by screening out catechin and epicatechin gallate from the tea extract.

Graphical Abstract

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分级介孔zif -8固定化酶微反应器-高效液相色谱法筛选绿茶中α-葡萄糖苷酶抑制剂

以热敏水凝胶为软模板合成了分级介孔ZIF-8 （HZIF-8）。锌离子（Zn2+）通过与水杨酸和三聚氰胺的金属离子配位，在室温下与2-甲基咪唑配位，均匀分散在水凝胶表面。对水杨酸和三聚氰胺进行热脱除后，得到HZIF-8， HZIF-8具有微孔和介孔双峰结构。采用原位扩散法将α-葡萄糖苷酶（α-Glu）包封在HZIF-8中，构建固定化α-Glu微反应器。利用傅里叶变换红外光谱（FT-IR）、电镜、x射线衍射（XRD）和布鲁诺尔-埃米特-泰勒（BET）分析对α-Glu固定前后HZIF-8的形态和结构特征进行了全面表征。结果表明，最佳固定时间为3 h， HZIF-8与α-Glu的理想质量比为5:2 （w/w）。在此条件下，α-Glu负载量达到82.09 μg/mg，固定化α-Glu微反应器经过7个循环后仍保持70%的酶活性。通过与高效液相色谱（HPLC）联用验证了HZIF-8-α-Glu微反应器在绿茶提取物中α-Glu抑制剂离线筛选中的应用。通过对茶提取物中儿茶素和表儿茶素没食子酸酯的筛选，验证了固定化α-Glu微反应器的应用。图形抽象

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.