Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, People's Republic of China; University of Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100049, People's Republic of China.
Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, People's Republic of China.
Talanta. 2017 Mar 1;164:548-555. doi: 10.1016/j.talanta.2016.12.028. Epub 2016 Dec 13.
As the close correlation between α-glucosidase inhibitors and the treatment of diabetes, in combination with capillary electrophoresis (CE), a method was developed to screen α-glucosidase inhibitors from traditional Chinese medicines (TCMs) by immobilizing α-glucosidase on magnetic nanoparticles. Such a magnetic immobilization would be beneficial for enzyme reusability, stability and separation. In this work, FeO nanoparticles were synthesized by solvothermal method. And the prepared nanoparticles were characterized by transmission electron microscopy (TEM), Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), size and zeta potential analysis. With the modification of chitosan (CS) and glutaraldehyde (GA), α-glucosidase was successfully immobilized on the magnetic nanoparticles. The pH and temperature endurance, storage stability and reusability of the immobilized α-glucosidase were studied and compared with those of the free one. With the magnetic immobilized α-glucosidase, the Michaelis-Menten constant (K) was calculated to be 0.85mM, and the inhibition constant (K) and half-maximal inhibitory concentration (IC) for acarbose were determined to be 7.37 and 13.69μM, respectively. Finally, the developed method was applied to screen α-glucosidase inhibitors from 18 TCMs.
由于 α-葡萄糖苷酶抑制剂与糖尿病治疗密切相关，本研究将 α-葡萄糖苷酶固定在磁性纳米粒子上，结合毛细管电泳（CE），从中药（TCM）中筛选 α-葡萄糖苷酶抑制剂。这种磁性固定有利于酶的重复使用、稳定性和分离。本工作采用溶剂热法合成了 FeO 纳米粒子。通过透射电子显微镜（TEM）、傅里叶变换红外（FT-IR）光谱、X 射线衍射（XRD）、粒径和 Zeta 电位分析对制备的纳米粒子进行了表征。通过壳聚糖（CS）和戊二醛（GA）的修饰，成功地将 α-葡萄糖苷酶固定在磁性纳米粒子上。研究了固定化 α-葡萄糖苷酶的 pH 和温度耐受性、储存稳定性和可重复使用性，并与游离酶进行了比较。用磁性固定化的 α-葡萄糖苷酶，计算得到米氏常数（K）为 0.85mM，阿卡波糖的抑制常数（K）和半最大抑制浓度（IC）分别为 7.37 和 13.69μM。最后，该方法被应用于从 18 种中药中筛选 α-葡萄糖苷酶抑制剂。
