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克隆和鉴定参与金银花绿原酸生物合成的一个编码对香豆酰辅酶 A 奎尼酸对香豆酰转移酶的 cDNA。

Cloning and characterization of a cDNA coding a hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase involved in chlorogenic acid biosynthesis in Lonicera japonica.

机构信息

Department of Biopharmaceutics, School of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China.

出版信息

Planta Med. 2010 Nov;76(16):1921-6. doi: 10.1055/s-0030-1250020. Epub 2010 Jun 10.

Abstract

The flower buds of Lonicera japonica are widely used in Chinese medicine for their anti-inflammatory properties. The reason why L. japonica has potent and significant effects is that it contains various active components, especially chlorogenic acid (CGA). In the present study, we isolated a hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (HQT) gene encoding a protein of 439 amino acids from L. japonica. The RT-PCR results showed that tissue distribution of HQT was in accordance with the pattern of CGA content. The corresponding cDNAs were expressed in Escherichia coli as fusion proteins. Recombinant HQT exhibited catalysis activity in esterification of quinic acid with caffeoyl-CoA to generate CGA. Purified proteins demonstrated similar kinetic properties as HQT isolated from other species. This work is of significance for further studies concerned with strengthening the efficacy of L. japonica by means of improved CGA content and exploring the biosynthesis of CGA in vitro.

摘要

金银花的花蕾在中药中被广泛应用,因其具有抗炎特性。金银花之所以具有强大而显著的功效,是因为它含有各种活性成分,特别是绿原酸(CGA)。在本研究中,我们从金银花中分离出一个编码 439 个氨基酸的羟基肉桂酰辅酶 A 奎宁酸羟基肉桂酰转移酶(HQT)基因。RT-PCR 结果表明,HQT 的组织分布与 CGA 含量的模式一致。相应的 cDNA 在大肠杆菌中表达为融合蛋白。重组 HQT 表现出催化奎宁酸与咖啡酰辅酶 A 酯化生成 CGA 的活性。纯化的蛋白质表现出与从其他物种中分离出的 HQT 相似的动力学特性。这项工作对于通过提高 CGA 含量来增强金银花的功效以及探索 CGA 的体外生物合成具有重要意义。

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