琯溪蜜柚柚皮苷纯化鉴定及其对胰脂肪酶的抑制作用  

Purification,Identification and Pancreatic Lipase Inhibitory Effect of Naringin from the Peel of Guanxi HoneyPomelo(Citrus grandis(L.)Osbeck)

作　　者：晏幸 肖文熙 欧丽明 上官宇晨 杜希萍[1,3,4] 李利君 胡阳[1,3,4] 倪辉 YAN Xing;XIAO Wenxi;OU Liming;SHANGGUAN Yuchen;DU Xiping;LI Lijun;HU Yang;NI Hui(College of Ocean Food and Biological Engineering,Jimei University,Xiamen 361021,China;College of Harbour and Coastal Engineering,Jimei University,Xiamen 361021,China;Key Laboratory of Food Microbiology and Enzyme Engineering Technology of Fujian Province,Xiamen 361021,China;Research Center of Food Biotechnology of Xiamen City,Xiamen 361021,China)

机构地区：[1]集美大学海洋食品与生物工程学院,福建厦门361021 [2]集美大学港口与海岸工程学院,福建厦门361021 [3]福建省食品微生物与酶工程重点实验室,福建厦门361021 [4]厦门市食品生物工程技术研究中心,福建厦门361021

出　　处：《食品科学》2022年第7期1-7,共7页Food Science

基　　金：国家自然科学基金联合基金项目(U1805235);福建省平和县现代农业产业园中央财政奖补资金项目(PH2020)。

摘　　要：为了探索琯溪蜜柚黄酮作为胰脂肪酶抑制剂的应用价值,本实验采用Sephadex LH-20葡聚糖柱层析、高效液相色谱-质谱联用、傅里叶变换红外光谱对琯溪蜜柚柚皮黄酮进行分离纯化及结构鉴定,并通过酶反应动力学和分子对接技术研究其对胰脂肪酶的抑制类型和分子间相互作用。高效液相色谱-质谱联用和傅里叶变换红外光谱分析结果表明,纯化后的琯溪蜜柚柚皮主要黄酮成分为柚皮苷;纯化鉴定的柚皮苷对胰脂肪酶半抑制浓度(half inhibitory concentration,IC_(50))为0.99μg/mL,抑制类型为可逆非竞争型,抑制常数Ki=0.58μg/mL;该黄酮与胰脂肪酶的结合自由能为-30.14 kJ/mol,相互间的分子作用力包括范德华力、疏水相互作用、氢键、π键等,结合区域位于酶催化活性中心之外的位点。本研究表明琯溪蜜柚的主要黄酮成分柚皮苷对胰脂肪酶具有强烈的抑制作用,为利用琯溪蜜柚黄酮开发高效安全的胰脂肪酶抑制剂提供了理论依据。In order to explore the potential of flavonoids from Guanxi honey pomelo as a pancreatic lipase inhibitor,flavonoids from the fruit’s peel were purified by Sephadex LH-20 column chromatography and identified by high performance liquid chromatography-mass spectrometry(HPLC-MS)and Fourier transform infrared(FTIR)spectroscopy.Moreover,the type of inhibition and the molecular interaction between the flavonoids and pancreatic lipase were studied by enzymatic reaction kinetics and molecular docking.The results of purification and identification showed the major flavonoid compound in Guanxi honey pomelo was naringin.The half-maximal inhibitory concentration(IC_(50))of naringin on pancreatic lipase was 0.99μg/mL.The inhibition was reversible non-competitive.The inhibition constant Ki was 0.58μg/mL.Molecular docking showed that the binding free energy between the flavonoid compound and the enzyme was-30.14 kJ/mol.They interacted with each other at a position outside the catalytic core via van der Waals force,hydrophobic interaction,hydrogen bond andπbond.This study revealed that naringin as one of the main flavonoids in Guanxi honey pomelo could strongly inhibit pancreatic lipase,which will provide a theoretical basis for the development of flavonoids from Guanxi honey pomelo as new and safe pancreatic lipase inhibitors.

关 键 词：琯溪蜜柚 柚皮苷 分离纯化 结构鉴定 胰脂肪酶 非竞争性抑制 

分 类 号：TS201.2[轻工技术与工程—食品科学]