Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
Anal Chim Acta. 2013 Jun 27;785:75-81. doi: 10.1016/j.aca.2013.04.058. Epub 2013 May 9.
A novel kind of immobilized enzyme affinity selection strategy based on hollow fibers has been developed for screening inhibitors from extracts of medicinal plants. Lipases from porcine pancreas were adsorbed onto the surface of polypropylene hollow fibers to form a stable matrix for ligand fishing, which was called hollow fibers based affinity selection (HF-AS). A variety of factors related to binding capability, including enzyme concentration, incubation time, temperature, buffer pH and ion strength, were optimized using a known lipase inhibitor hesperidin. The proposed approach was applied in screening potential lipase bound ligands from extracts of lotus leaf, followed by rapid characterization of active compounds using high performance liquid chromatography-mass spectrometry. Three flavonoids including quercetin-3-O-β-D-arabinopyranosyl-(1→2)-β-D-galactopyranoside, quercetin-3-O-β-D-glucuronide and kaempferol-3-O-β-d-glucuronide were identified as lipase inhibitors by the proposed HF-AS approach. Our findings suggested that the hollow fiber-based affinity selection could be a rapid and convenient approach for drug discovery from natural products resources.
一种新型的基于中空纤维的固定化酶亲和选择策略已被开发用于从药用植物提取物中筛选抑制剂。猪胰腺脂肪酶被吸附到聚丙烯中空纤维的表面上，形成用于配体捕捞的稳定基质，这被称为中空纤维亲和选择（HF-AS）。使用已知的脂肪酶抑制剂橙皮苷优化了与结合能力相关的多种因素，包括酶浓度、孵育时间、温度、缓冲液 pH 值和离子强度。该方法已应用于从荷叶提取物中筛选潜在的脂肪酶结合配体，然后使用高效液相色谱-质谱法快速表征活性化合物。通过所提出的 HF-AS 方法，鉴定出三种类黄酮，包括槲皮素-3-O-β-D-阿拉伯吡喃糖苷基-(1→2)-β-D-半乳糖吡喃糖苷、槲皮素-3-O-β-D-葡萄糖醛酸苷和山奈酚-3-O-β-D-葡萄糖醛酸苷，它们是脂肪酶抑制剂。我们的研究结果表明，基于中空纤维的亲和选择可能是从天然产物资源中发现药物的一种快速便捷的方法。
