茶叶-葡萄糖苷酶亲和层析纯化与性质研究

doi:10.13305/j.cnki.jts.2005.01.004

茶叶科学 ›› 2005, Vol. 25 ›› Issue (1): 16-22.doi: 10.13305/j.cnki.jts.2005.01.004

茶叶-葡萄糖苷酶亲和层析纯化与性质研究

张正竹¹, 宛晓春¹, 坂田完三²

1. 安徽农业大学农业部茶叶生物化学与生物技术重点实验室,安徽合肥 230036;
2. 京都大学化学研究所,日本京都 611-0011

出版日期:2005-03-25 发布日期:2019-09-16
作者简介:张正竹（1969— ）,男,博士,副教授,主要从事茶叶生物化学与分子生物学研究。
基金资助:
国家攀登计划（国科基字[1999]045号）、安徽省教育厅自然科学研究项目（2003kj130）与教育部留学回国人员科研启动基金项目内容之一

Purification by Affinity Chromatography and Characterization of Tea Leaf -Glucosidase

ZHANG Zheng-zhu¹, WAN Xiao-chun¹, KANZO Sakata²

1 Key Laboratory of Tea Biochemistry & Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, Chian;
2. Institute for Chemical Research, Kyoto University, Kyoto 611-0011, Japan

Online:2005-03-25 Published:2019-09-16

摘要/Abstract

摘要： 以化学合成的β-葡萄糖苷酶的抑制剂β-葡萄糖脒为配体,通过N-糖苷键交联到支持物上,使糖脒N-糖苷键一侧的氨基保持带正电荷的活化状态,合成了一种β-葡萄糖苷酶的亲和层析树脂。这种合成的亲和层析树脂在弱酸条件下非常稳定,能选择性地吸附β-葡萄糖苷酶,并能多次重复使用。利用这种亲和层析树脂,在pH6和pH5条件下,从薮北种茶树叶片中纯化出两种分子量分别为63和75βkDa的β-葡萄糖苷酶单体酶。以对硝基苯酚β-葡萄糖苷为底物,研究了两种酶的性质。TLC分析结果表明,这两种β-葡萄糖苷酶都能水解(Z)-3-己烯醇、苯甲醇和苯乙腈的β-D-葡萄糖苷。

关键词: 茶学, β-葡萄糖苷酶, 茶叶香气, 亲和层析, 薮北种, β-葡萄糖脒

Abstract: Two kinds of β-Glucosidases from Camellia sinensis var. sinensis cv. Yabukita were purified by a sophisticate application of affinity chromatography. An affinity adsorbent for β-Glucosidases has been prepared with a positively charged β-glucosylamidine as a ligand in which a glycon moiety is connected via a N-glycoside linkage. The synthesized affinity adsorbent is quite stable under mildly acidic conditions (pH5~6) and can be used up to 20 times over three months without significant affect on the column performance. Two tea leaf β-Glucosidases have been purified to be monomers with molecular mass of 63 and 75 kDa by affinity chromatography at pH6.0 and pH5.0, respectively. The extensive characterization of tea leaf β-Glucosidasse has been reported. Tea aroma precursors such as benzyl, (Z)-3-hexenyl and D-mandelonitrile β-D-glucopyranosides were hydrolyzed by the purified two β-Glucosidases to liberate tea aroma.

Key words: Tea science, β-Glucosidase, Tea aroma, Affinity chromatography, Camellia sinensis var, Sinensis cv, Yabukita, β-Glucosylamidine

中图分类号:

S571.1
Q50

张正竹, 宛晓春, 坂田完三. 茶叶-葡萄糖苷酶亲和层析纯化与性质研究[J]. 茶叶科学, 2005, 25(1): 16-22. doi: 10.13305/j.cnki.jts.2005.01.004.

ZHANG Zheng-zhu, WAN Xiao-chun, KANZO Sakata. Purification by Affinity Chromatography and Characterization of Tea Leaf -Glucosidase[J]. Journal of Tea Science, 2005, 25(1): 16-22. doi: 10.13305/j.cnki.jts.2005.01.004.

参考文献 7

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茶叶-葡萄糖苷酶亲和层析纯化与性质研究

Purification by Affinity Chromatography and Characterization of Tea Leaf -Glucosidase

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