| 茶树酯型儿茶素水解酶鉴定及其检测体系的建立 |
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| 引用本文： | 聂志银,刘亚军,刘莉,高丽萍,夏涛. 茶树酯型儿茶素水解酶鉴定及其检测体系的建立[J]. 茶叶科学, 2011, 31(5): 439-446. DOI: 10.13305/j.cnki.jts.2011.05.014 |
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| 作者姓名： | 聂志银 刘亚军 刘莉 高丽萍 夏涛 |
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| 作者单位： | 1. 安徽农业大学生命科学学院,安徽 合肥 230036; 2. 安徽农业大学教育部茶叶生物化学与生物技术重点实验室,安徽 合肥 230036 |
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| 基金项目： | 国家自然科学基金(30771755,30972401); 安徽省自然科学基金(090411006); 国家重点基础研究发展计划“973”前期项目(2007CB116211) |
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| 摘 要： | 本试验利用体外酶学方法,结合薄层层析（TLC）、高效液相色谱（HPLC）和液相色谱-串联质谱（LC-MS/MS）分析,首次从茶树中检测到活性较高的酯型儿茶素水解酶（Galloylated Catechins Hydrolase,GCH）的存在。在酯型儿茶素水解酶催化下,酯型儿茶素发生水解反应,生成没食子酸（GA）和非酯型儿茶素。试验确立了酯型儿茶素水解酶的最适检测体系,在2.5mL反应体系中包含0.2mmol/L酯型儿茶素、2.4mmol/L抗坏血酸、粗酶提取液若干（含0.1mg酶蛋白）和0.1mol/L磷酸缓冲液（pH6.5）,在30℃下,反应30min。此外,试验利用硫酸铵分级沉淀、阴离子交换层析和凝胶过滤层析对该酶进行了初步纯化。
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| 关 键 词： | 茶树 酯型儿茶素水解酶 鉴定 检测体系 |
| 收稿时间： | 2011-02-25 |
Identification and Reaction Assay of Galloylated Catechins Hydrolase in Tea Plant |
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| NIE Zhi-yin,LIU Ya-jun,LIU Li,GAO Li-ping,XIA Tao. Identification and Reaction Assay of Galloylated Catechins Hydrolase in Tea Plant[J]. Journal of Tea Science, 2011, 31(5): 439-446. DOI: 10.13305/j.cnki.jts.2011.05.014 |
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| Authors: | NIE Zhi-yin LIU Ya-jun LIU Li GAO Li-ping XIA Tao |
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| Affiliation: | 1. School of Biology Science, Anhui Agricultural University, Hefei 230036, China; 2. Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education, Anhui Agricultural University, Hefei 230036, China |
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| Abstract: | In this experiment,high activity of galloylated catechins hydrolase(GCH) was detected existing in tea plant(Camellia sinensis(L.) O.Kuntze) by enzymology analysis in vitro,combining thin layer chromatography(TLC),high performance liquid chromatography(HPLC),and liquid chromatography with tandem mass spectrometry(LC-MS/MS) analysis.The galloylated catechins could be hydrolyzed to ungalloylated catechins and gallic acid(GA) with the GCH action.The optimum reaction assay of GCH has been established.The 2.5 mL ... |
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| Keywords: | tea plant(Camellia sinensis(L.) O.Kuntze) galloylated catechins hydrolase identification reaction assay |
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