Enzymatic synthesis of new terpenes from terpene synthase substrate analogs

从萜烯合酶底物类似物酶法合成新萜烯

• 批准号: 513548540
• 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
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/513548540?language=en
• 关键词: Enzymatic; synthesis; new; terpenes; terpene

This project aims at the synthesis of substrate analogs of terpene synthases and their enzymatic conversions. The planned structural modifications include substrate analogs with shifted double bonds for an expected change of reactivity, with hydrogenated double bonds (blocked reactivity, expectedly leading to derailment products), and removed methyl groups (buffered reactivity). Furthermore, a series of substrate analogs with skeletal extensions by one to four carbons will be synthesised that allow to study functional switches between diterpene and sesterterpene, or between sesterterpene and triterpene synthases. All compounds obtained in this project will be isolated and their structures will be elucidated by spectroscopic methods. Absolute configurations will be determined through a stereoselective deuteration strategy previously established in our laboratory. If for steric reasons certain enzyme-substrate combinations fail to work, enzyme variants with enlarged active site cavities will be obtained by site-directed mutagenesis. DFT calculations in collaboration with Prof. Dr. Bernd Goldfuss (University of Cologne) will add to a deeper understanding of interesting cyclisation mechanisms. Overall, the aim of this project is to broadly show the applicability of terpene synthases in the conversion of substrate analogs to make new terpenes accessible that cannot be obtained from the natural substrates.

本项目旨在合成萜烯脱氢酶的底物类似物及其酶促转化。计划的结构修饰包括具有移位双键的底物类似物，用于预期的反应性变化，具有氢化双键（阻断反应性，导致脱轨产物的氢化），以及去除甲基（缓冲反应性）。此外，将合成一系列具有一到四个碳的骨架延伸的底物类似物，其允许研究二萜和二萜之间或二萜和三萜脱氢酶之间的功能开关。本项目中获得的所有化合物将被分离，并通过光谱方法鉴定其结构。绝对构型将通过我们实验室先前建立的立体选择性氘化策略来确定。如果由于空间原因某些酶-底物组合不能起作用，则通过定点诱变获得具有扩大的活性位点空腔的酶变体。与Bernd Goldfuss教授（科隆大学）合作进行的DFT计算将加深对有趣的环化机制的理解。总的来说，该项目的目的是广泛地显示萜烯脱氢酶在底物类似物转化中的适用性，以使不能从天然底物中获得的新萜烯可获得。