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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

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【作者】 屠幼英童启庆赵芹渡边修治Fleishmann.P

【Author】 Tu You_ying 1,Tong Qi_qing 1,Zhao Qin 1, Watanabe.N 2,Fleischmann.P 2 (1. Department of Tea Science, Zhejiang University, Hangzhou 310029, China; 2. Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan)

【机构】 浙江大学茶学系静冈大学农学部静冈大学农学部 浙江杭州310029浙江杭州310029静冈422-8529日本日本

【摘要】 茉莉花芳樟醇 βD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CMToyopearl65 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SephacrylS2 0 0柱分离系统纯化出了LGA ,LGA蛋白的SDSPAGE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 βD喃葡萄糖苷分解释放芳樟醇

【Abstract】 The acetone powder of linalyl βD 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 CMToyopearl 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 S200 column and eluted by the same buffer. A part of fraction containing LGA activity was assayed by SDSPAGE 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】 茉莉花LGA纯化分子量
【Key words】 Jasminum sambic AitsLGAPurificationMolecular weight
【基金】 国家自然科学基金 (39570 4 31 );国际交流合作资助项目
  • 【文献出处】 茶叶科学 ,Journal of Tea Science , 编辑部邮箱 ,2001年02期
  • 【分类号】Q946.5
  • 【被引频次】8
  • 【下载频次】239
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