Structural differences of hydroxycinnamoyl transferases in Lamiaceae - Part 1: The structure of rosmarinic acid synthase

唇形科羟基肉桂酰转移酶的结构差异 - 第 1 部分：迷迭香酸合酶的结构

• 批准号: 43660071
• 负责人: Professorin Dr. Maike Petersen
• 金额: --
• 依托单位: Institut für Pharmazeutische Biologie und Biotechnologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/43660071?language=en
• 关键词: Structural; differences; hydroxycinnamoyl; transferases; Lamiaceae

Hydroxycinnamoyltransferases (HCTs) are important enzymes in plant metabolism, in this project we are interested in the HCTs from the BAHD acyltransferase superfamily, especially within one phylogenetic clade. These HCTs transfer a hydroxycinnamoyi moiety from coenzyme A to an acceptor substrate such as shikimate, quinate or hydroxyphenyllactate which finally results in the formation of compounds like monolignols, chlorogenic acid and rosmarinic acid, respectively. The HCTs involved in these biosyntheses show high homology. Despite their importance for plant metabolism, structural information on these enzymes is scarce, but it is urgently needed to explain the diverse substrate acceptance that was reported for HCTs as well as the formation of esters and amides by the same enzyme. From an evolutionary point of view, the changes in the active centres of HCTs resulting in different substrate preferences are interesting. During the first application period we have tried to crystallise rosmarinic acid synthase (RAS) from Coleus blumei - unfortunately without sufficient success. We now want to apply our ample experiences with handling HCTs for a novel attempt to get insight into the structure of HCTs. Modelling and substrate docking will be used besides crystallisation. Further parts of this project are site-directed mutagenesis and construction of chimeric proteins of different HCTs with subsequent testing of their substrate acceptance.

羟基肉桂基转移酶（Hydroxycinnamoyltransferases, hct）是植物代谢过程中重要的酶，本项目对BAHD酰基转移酶超家族的hct感兴趣，特别是在一个系统发育分支中。这些hct将辅酶a的羟基肉桂酸片段转移到一个受体底物上，如莽草酸盐、奎宁酸盐或羟基苯基乳酸盐，最终形成化合物，如单木脂醇、绿原酸和迷迭香酸。参与这些生物合成的hct具有高度的同源性。尽管它们对植物代谢很重要，但关于这些酶的结构信息很少，但迫切需要解释hct对不同底物的接受，以及由同一酶形成酯和酰胺。从进化的角度来看，hct活性中心的变化导致不同的底物偏好是有趣的。在第一个申请期间，我们试图从蓝花中结晶迷迭香酸合成酶(RAS) -不幸的是没有足够的成功。我们现在想运用我们在处理hct方面的丰富经验，进行一种新的尝试，以深入了解hct的结构。除了结晶之外，还将使用建模和衬底对接。该项目的进一步部分是位点定向诱变和不同hct嵌合蛋白的构建，并随后测试其底物接受性。