Thermoplasma volcanium来源β-葡萄糖苷酶的酶学性质分析  

Enzymatic Properties Analysis ofβ-glucosidase from Thermoplasma volcanium

作　　者：刘阳 王丹[1,2] 纪锐[1,2] 顾少华[1,2] 李继喜[1,2] 季朝能[1,2] LIU Yang;WANG Dan;JI Rui;GU Shaohua;LI Jixi;JI Chaoneng(School of Life Sciences,Fudan University,Shanghai 200438,China;State Key Laboratory of Genetic Engineering,Shanghai 200438,China)

机构地区：[1]复旦大学生命科学学院,上海200438 [2]复旦大学遗传工程国家重点实验室,上海200438

出　　处：《复旦学报（自然科学版）》2021年第5期636-645,共10页Journal of Fudan University：Natural Science

基　　金：国家重点研发计划(2016YFA0500600);上海市科学技术委员会项目(19DZ2282100)。

摘　　要：β-葡萄糖苷酶(BGL)是催化纤维素产生葡萄糖过程中的关键酶,在生物能源利用和食品领域发挥重要作用.本研究克隆了来源于Thermoplasma volcanium的β-葡萄糖苷酶(BGL)基因,并在大肠杆菌中进行表达,经纯化获得了Tv-BGL.酶学性质分析发现,Tv-BGL在pH6.0和80℃时发挥最佳催化活力,以对硝基苯基-β-D-吡喃葡萄糖苷(pNPGlu)为底物时的K_(m)和V_(max)分别为0.7254 mmol/L和614.60 U/mg,K_(cat)/K_(m)=7.79×10^(5) mol^(-1)·L·s^(-1),以纤维二糖(Cellobiose)为底物时的K_(m)和V_(max)分别为55.63 mmol/L和636.06 U/mg,Kcat/Km=1.06×10^(4) mol^(-1)·L·s^(-1).另外,该酶具有较好的热稳定性,有机溶剂、离子以及葡萄糖耐受性.β-Glucosidase(BGL)is a critical enzyme in the process of catalyzing the production of glucose from cellulose and plays an essential role in the use of bioenergy and food.In this study,theβ-glucosidase(BGL)gene derived from Thermoplasma volcanium was cloned and expressed in E.coli,and protein was obtained after purification.The analysis of enzymatic properties found that Tv-BGL exerts the highest catalytic activity at pH 6.0 and 80℃.The K_(m) and V_(max) with substrate p-nitrophenyl-β-D-glucopyranoside(pNPGlu)are 0.7254 mmol/L and 614.60 U/mg,respectively,and the enzyme has K_(cat)/K_(m)=7.79×10^(5) mol^(-1)·L·s^(-1),K_(m) and V_(max) when using Cellobiose as the substrate are 55.63 mmol/L and 636.06 U/mg,respectively,Kcat/Km=1.06×10^(4) mol^(-1)·L·s^(-1).In addition,the enzyme has good thermal stability,organic solvent tolerance,ion tolerance,and glucose tolerance,which are beneficial to industrial applications.

关 键 词：Β-葡萄糖苷酶 糖苷水解酶 热稳定 葡萄糖耐受 工业应用 

分 类 号：Q518.3[生物学—生物化学]