Biosynthesis and utilisation of SAM diastereomers as tools for the characterisation of SAM-dependent enzymes and product diversification

SAM 非对映异构体的生物合成和利用作为表征 SAM 依赖性酶和产品多样化的工具

• 批准号: 530629952
• 负责人: Professorin Dr. Jennifer Andexer
• 金额: --
• 依托单位: Institut für Pharmazeutische Wissenschaften
• 依托单位国家: 德国
• 项目类别: Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/530629952?language=en
• 关键词: Biosynthesis; utilisation; SAM; diastereomers; tools

(S,S)-S-adenosylmethionine (SAM) contains six stereocentres, four of those are fixed through the ribose configuration of the adenosine moiety. Another stereocentre is defined through the alpha-carbon configuration of methionine, and the last one at the sulfonium centre is introduced by methionine adenosyltransferase (MAT) during SAM biosynthesis. Especially the sulfonium epimer is of relevance, as it is an inhibitor for methyltransferases (MTs) and develops non-enzymatically, especially in in vitro applications. The alpha-carbon epimer is similarly interesting, as it might be present in racemic mixtures of methionine analogues used for product diversification. In addition, both types of SAM diastereomers have the potential to lead to different products in reactions catalysed by SAM-dependent enzymes. This project focuses on the biocatalytic synthesis of the corresponding four SAM diastereomers using MATs, chlorinases and SAM hydrolases, and their interaction with different types of SAM-dependent enzymes, covering MTs, radical SAM enzymes, and enzymes using the SAM aminocarboxypropyl (ACP) group. After a systematic analysis of different types of SAM-dependent enzymes present in the planned FOR with the SAM diastereomers regarding acceptance, substrate binding and product outcome, the suitability of SAM diastereomers as strategy for the development of orthogonal pathways in vivo will be explored.

（S，S）-S-腺苷蛋氨酸（SAM）含有六个立体中心，其中四个通过腺苷部分的核糖结构固定。另一个立体中心是通过蛋氨酸的α -碳结构定义的，最后一个在磺胺中心是由蛋氨酸腺苷转移酶（MAT）在SAM生物合成过程中引入的。尤其是表聚体磺酸，因为它是甲基转移酶（MTs）的抑制剂，并且是非酶性的，特别是在体外应用中。α -碳外聚体同样有趣，因为它可能存在于用于产品多样化的蛋氨酸类似物的外消旋混合物中。此外，两种类型的SAM非对映体都有可能在SAM依赖酶催化的反应中产生不同的产物。本项目重点研究了四种SAM非对映体的生物催化合成，包括mat、氯酶和SAM水解酶，以及它们与不同类型SAM依赖酶的相互作用，包括MTs、自由基SAM酶和使用SAM氨基羧基（ACP）基的酶。在系统分析了不同类型的SAM依赖酶与SAM非对映体在接受、底物结合和产物结果方面的关系后，将探索SAM非对映体作为开发体内正交途径策略的适用性。