茉莉花芳樟醇β-D吡喃葡萄糖苷酶的分离与纯化  

The Isolation and Purification of LGA from Jasminum sambic Ait

作　　者：屠幼英[1] 童启庆[1] 赵芹[1] 渡边修治[2] Fleishmann.P 

机构地区：[1]浙江大学茶学系,浙江杭州310029 [2]静冈大学农学部

出　　处：《茶叶科学》2001年第2期144-147,共4页Journal of Tea Science

基　　金：国家自然科学基金 (39570 4 31 );国际交流合作资助项目

摘　　要：茉莉花芳樟醇 β_D吡喃葡萄糖苷酶 (LGA)丙酮粉经过pH6 0柠檬酸缓冲液萃取、 3 0 %和 80 %硫酸铵分步沉淀 ,以及CM_Toyopearl65 0M柱层析 ,用 0 - 0 5MNaCl柠檬酸缓冲液梯度洗脱酶蛋白 ,最后用快速蛋白液相色谱 (FPLC)的SephacrylS_2 0 0柱分离系统纯化出了LGA ,LGA蛋白的SDS_PAGE图谱在分子量 1 4 4万道尔顿处显示了一条谱带。它可以催化芳樟基The acetone powder of linalyl β_D_ pyranoglucosidase(LGA) from Jasminum sambic flower was homogenized with 0 1 M pH6 0 citrate buffer and centrifuged, the supernatant was filtrated by 0 2 μm membrane filtration, then make the fractional precipitation of crude enzyme solution with 30% and 80% (NH4) 2SO 4 After centrifugation, the precipitate was dissolved and dialyzed in the same buffer The dialyzed enzyme was added on CM_Toyopearl 650 M column and eluted by a linear gradient of 0-0 5 M NaCl in the citrate buffer. The fraction containing LGA activity was collected, and freeze dried and dialyzed, the dialyzed solution was placed on a Sephacryl S_200 column and eluted by the same buffer. A part of fraction containing LGA activity was assayed by SDS_PAGE with CBB staining method, the result of electrophoresis showed one band on the gel, the molecular weight (WM) of this protein was about 14 4kD.The other part containing LGA activity showed two bands, MW was 96 0kD and 30 0kD respectively.

关 键 词：芳樟醇β-D吡喃葡萄糖苷酶 分离 纯化 LGA 茉莉花 

分 类 号：Q946.5[生物学—植物学]