Synthesis of isotopically labelled terpene precursors for structure elucidation of terpenes and investigation of mechanistic aspects of their biosynthesis

合成同位素标记的萜烯前体，用于萜烯的结构阐明及其生物合成的机制方面的研究

• 批准号: 271721844
• 负责人: Professor Dr. Jeroen Dickschat
• 金额: --
• 依托单位: Kekulé-Institut für Organische Chemie und Biochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/271721844?language=en
• 关键词: Synthesis; isotopically; labelled; terpene; precursors

In this project several methylated analogs of the terpene monomers dimethylallyl pyrophosphate (DMAPP) and isopentenyl pyrophosphat (IPP) will be synthesised. The obtained compounds will be coupled enzymatically to geranyl diphosphate (GPP), farnesyl diphosphate (FPP) or IPP using an FPP or GGPP synthase to obtain methylated analogs of FPP and geranylgeranyl diphosphate (GGPP) with one additional methyl group. Subsequently, also the incorporation of more than one additional methyl group will be tried. The products shall be dephosphorylated by a phosphatase, isolated and their structures will be elucidated by NMR spectroscopy. The obtained methylated acyclic terpene precursors will be converted into cyclic products using various terpene synthases. The formed terpenes will be isolated and their structures will be elucidated. Through this approach the naturally accessible structural space will be significantly expanded and our understanding of the plasticity of terpene synthases will be deepened. Furthermore, an already existing library of 13C-labelled terpene precursors will be extended to make all singly 13C-labelled isotopomers of methylated terpene analogs accessible by enzyme reactions. With the labelled compounds the cyclisation mechanisms of artificial terpenes and their EI-MS fragmentation mechanisms can be studied.

在本项目中，将合成萜烯单体二甲基烯丙基焦磷酸（DMAPP）和异戊烯基焦磷酸（IPP）的几种甲基化类似物。使用FPP或GGPP合酶将获得的化合物酶促偶联至香叶基二磷酸（GPP）、法尼基二磷酸（FPP）或IPP，以获得FPP和具有一个额外甲基的香叶基香叶基二磷酸（GGPP）的甲基化类似物。随后，还将尝试引入多于一个另外的甲基。产物应通过磷酸酶脱磷酸化，分离，并通过NMR光谱法解析其结构。所获得的甲基化无环萜烯前体将使用各种萜烯脱氢酶转化为环状产物。形成的萜烯将被分离，其结构将被阐明。通过这种方法，自然可及的结构空间将显着扩大，我们的萜烯脱氢酶的可塑性的理解将加深。此外，已经存在的13 C-标记的萜烯前体库将被扩展，使所有单一的13 C-标记的同位素异构体的甲基化萜烯类似物可通过酶反应。利用标记化合物可以研究人工萜类化合物的环化机理及其EI-MS裂解机理。