黑曲霉α-L-鼠李糖苷酶与柚皮苷分子动力模拟研究  

Molecular Dynamics Investigations of Aspergillus nigerα-L-rhamnosidase with Naringin

作　　者：巩建业 吴喆瑜 刘嘉男 廖辉 李利君 倪辉 GONG Jianye;WU Zheyu;LIU Jianan;LIAO Hui;LI Lijun;NI Hui(College of Food and Biological Engineering,Jimei University,Xiamen 361021,China;Fujian Provincial Key Laboratory of Food Microbiology and Engineering,Xiamen 361021,China)

机构地区：[1]集美大学食品与生物工程学院,福建厦门361021 [2]福建省食品微生物与酶工程重点实验室,福建厦门361021

出　　处：《食品与生物技术学报》2019年第12期66-72,共7页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(31371751);福建省农业高校产学研合作重大项目(2014N5010)。

摘　　要：α-L-鼠李糖苷酶能够水解柚皮苷生产普鲁宁和L-鼠李糖,可用于柑橘类果汁的脱苦处理。对柚皮苷和黑曲霉α-L-鼠李糖苷酶进行分子对接,采用分子动力模拟和MM-PBSA结合的方法计算对接复合物的结合自由能,并分析了分子间相互作用以及各个残基对结合自由能的贡献。结果表明范德华作用力是黑曲霉α-L-鼠李糖苷酶与柚皮苷结合的主要驱动力,库伦电荷作用和非极性溶剂化能对结合贡献较小。Trp236、Ala340、Ile462、Phe461、Tyr516、Val522、Trp528等是黑曲霉α-L-鼠李糖苷酶和柚皮苷结合过程中形成疏水作用的重要氨基酸残基。在分子动力模拟平衡后的氢键分析中,黑曲霉α-L-鼠李糖苷酶Ser286、Phe465、Pro520残基和柚皮苷共形成了3个稳定的氢键。本研究分析了黑曲霉α-L-鼠李糖苷酶与柚皮苷结合的驱动力,以及结合过程中重要氢键及疏水性残基,为蛋白质工程改造α-L-鼠李糖苷酶提供了靶位。α-L-Rhamnosidase generates prunin and L-rhamnose by hydrolyzing naringin,which can be used to debittering citrus juices.In the present study,theα-L-rhamnosidase,previously identified from Aspergillus niger JMU-TS528,was docked with naringin and the binding energy was calculated by molecular dynamics simulation and MM-PBSA method.The results of binding energy indicate that var der Waalsforce is the main driving force for enzyme-substrate combination;whereas the electrostatic interaction and non-polar solvents contribute less.In addition,the critical amino acid residues for hydrophobic effect are Trp236,Ala340,Ile462,Phe461,Tyr516,Val522 and Trp528.Ser286,Phe465 and Pro520 are important amino acid residues which could form steady hydrogen bonds with naringin.The key amino acid residues identified in the study provides valuable targeting sites for remouldingα-L-rhamnosidase in protein engineering.

关 键 词：α-L-鼠李糖苷酶 柚皮苷 分子对接 分子动力模拟 MM-PBSA 

分 类 号：Q816[生物学—生物工程]