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利用重组蛋白分析植物UDP依赖性糖基转移酶以鉴定其挥发性底物

Analyses of Plant UDP-Dependent Glycosyltransferases to Identify Their Volatile Substrates Using Recombinant Proteins.

作者信息

Kamiyoshihara Yusuke, Tieman Denise M, Klee Harry J

机构信息

Horticultural Sciences Department, University of Florida, Building 885, Gainesville, FL, 32611-0690, USA.

出版信息

Methods Mol Biol. 2016;1363:199-207. doi: 10.1007/978-1-4939-3115-6_16.

Abstract

Glycosylation is one of major modifications for plant secondary metabolites. In the case of volatile compounds, glycosylation makes them nonvolatile and odorless. Identification of UDP-dependent glycosyltransferases responsible for volatile glycosylation is essential to understand the regulatory mechanism of volatile release from plant tissues. Here, we describe an efficient protocol to find possible combinations of volatiles/glycosyltransferases using tomato (Solanum lycopersicum) enzymes expressed in Escherichia coli. The presented method requires a basic gas chromatography system and conventional laboratory tools.

摘要

糖基化是植物次生代谢产物的主要修饰方式之一。就挥发性化合物而言,糖基化使其变为非挥发性且无气味。鉴定负责挥发性化合物糖基化的UDP依赖性糖基转移酶对于理解植物组织中挥发性物质释放的调控机制至关重要。在此,我们描述了一种高效的方法,用于利用在大肠杆菌中表达的番茄(Solanum lycopersicum)酶来寻找挥发性物质/糖基转移酶的可能组合。所介绍的方法需要一个基本的气相色谱系统和常规实验室工具。

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