将吲哚胺2,3-双加氧酶1固定在磁性纳米颗粒上用于筛选咖啡中的抑制剂。
Indoleamine 2,3 dioxygenase 1 immobilization on magnetic nanoparticles for screening inhibitors from coffee.
作者信息
Liu Zhuoting, Ping Yunfang, Zhang Lumei, Zhang Jingran, Van Schepdael Ann, Wang Xu
机构信息
State Key Laboratory of Food Nutrition and Safety, College of Food Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China.
SCIEX, Analytical Instrument Trading Co., Ltd, Beijing 100015, China.
出版信息
Food Chem X. 2023 Jan 30;17:100591. doi: 10.1016/j.fochx.2023.100591. eCollection 2023 Mar 30.
In this study, a ligand fishing method was developed to screen potential indoleamine 2,3-dioxygenase 1 (IDO1) inhibitors from coffee extracts by immobilization of IDO1 enzyme on amino-modified magnetic nanoparticles combined with UHPLC-Q-TOF-MS/MS analysis. Parameters including enzyme concentration, immobilization time, the pH of glutaraldehyde and the amount of magnetic nanoparticles were optimized. The results indicated that immobilized IDO1 could be reused 5 times and was stable during storage for 7 days. Several IDO1 ligands were captured by incubating immobilized IDO1 with coffee extract, of which 10 showed an obvious difference comparing to non-conjugated bare nanoparticles. inhibitory activity was further performed by CE analysis, in which ferulic acid and chlorogenic acid had better IDO1 inhibitory activity, with IC value of 113.7 μM and 307.5 μM. These results demonstrate that this method provides an effective platform for identifying and screening IDO1 inhibitors from natural products.
在本研究中，开发了一种配体垂钓法，通过将吲哚胺2,3-双加氧酶1（IDO1）固定在氨基修饰的磁性纳米颗粒上，并结合超高效液相色谱-四极杆飞行时间串联质谱（UHPLC-Q-TOF-MS/MS）分析，从咖啡提取物中筛选潜在的IDO1抑制剂。对酶浓度、固定化时间、戊二醛pH值和磁性纳米颗粒用量等参数进行了优化。结果表明，固定化的IDO1可重复使用5次，且在储存7天期间保持稳定。通过将固定化的IDO1与咖啡提取物孵育捕获了几种IDO1配体，其中10种与未结合的裸纳米颗粒相比有明显差异。通过毛细管电泳（CE）分析进一步测定抑制活性，其中阿魏酸和绿原酸具有较好的IDO1抑制活性，IC值分别为113.7 μM和307.5 μM。这些结果表明，该方法为从天然产物中鉴定和筛选IDO1抑制剂提供了一个有效的平台。
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