Analysis of alpha-amylase inhibitor from corni fructus by coupling magnetic cross-linked enzyme aggregates of alpha-amylase with HPLC-MS

作者：Liu, Liangliang; Cen, Yin; Liu, Fang; Yu, Jingang; Jiang, Xinyu; Chen, Xiaoqing^*

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2015, 995: 64-69.

DOI：10.1016/j.jchromb.2015.05.016

• alpha-Amylase
• Corni fructus
• Cross-linked enzyme aggregates
• HPLC-MS
• Magnetic

As a carrier-free immobilization strategy, magnetic cross-linked enzyme aggregates (MCLEAs) showed improved enzyme activity, stability and magnetic response. In this study, MCLEAs of a-amylase (MCLEAs amylase) was prepared under optimized conditions and characterized with scanning electron microscope and vibrating sample magnetometer. The prepared MCLEAs amylase showed an amorphous structure and the saturation magnetization was 33.5 emu/g, which was sufficient for magnetic separation. Then MCLEAs amylase coupled with high performance liquid chromatography mass spectrometry (HPLC MS) was utilized to screen and identify a-amylase inhibitors from ethyl acetate extract of corni fructus. The experiment conditions were optimized. At the optimum conditions (incubation time: 10 min, pH: 7.0 and temperature: 20 degrees C), querciturone was successfully screened and identified with weak nonspecific binding. The screening result was verified by inhibition assays and the IC50 value of querciturone was 22.5 mu g/mL. This method provided a rapid way to screen active compounds from natural products.

• 出版日期2015-7-15
• 单位中南大学; 中国农业科学院