A Chemoenzymatic Approach to Accessing Novel Isoprenoid Scaffolds

获取新型类异戊二烯支架的化学酶方法

• 批准号: 10582364
• 负责人: Shanteri Singh
• 金额: $ 14.06万
• 依托单位: UNIVERSITY OF OKLAHOMA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10582364
• 关键词: Acids; Active Sites; Address; Alcohols; Award; Cannabinoids; Cells; Collaborations; Coupled; Coupling; Diphosphates; Engineering; Environment; Enzymes; Escherichia coli; Farnesol; Funding; Generations; Geranyltranstransferase; Goals; Human; In Situ; In Vitro; Industry; Isoprene; Kinetics; Knowledge; Libraries; Medicine; Metabolic; Metabolic Pathway; Methionine; Methodology; Methods; Methyltransferase; Minnesota; Modernization; Monitor; Mutagenesis; Natural Products; Nature; North Carolina; Parents; Pathway interactions; Phosphotransferases; Process; Production; Reaction; S-Adenosylmethionine; Source; Squalene; Squalene Synthetase; Structure; Structure-Activity Relationship; System; Technology; Therapeutic; Universities; Variant; Work; analog; analytical method; base; catalyst; chemical synthesis; combinatorial; farnesyl pyrophosphate; high throughput screening; in vivo; inorganic phosphate; isopentenyl pyrophosphate; isoprenoid; metabolic engineering; methionine adenosyltransferase; non-Native; novel; prenyl; scaffold; synthetic biology

Summary of the funded parent award (1R01GM138800-01A1). Isoprenoids represent a diverse class of compounds with a broad range of applications in medicine and industry. Their extraction from natural sources is both challenging and potentially harmful to the environment, while the enormous structural complexity of many isoprenoids makes traditional chemical synthesis nontrivial. Modern metabolic engineering and synthetic biology approaches have overcome some of these difficulties, but issues related to metabolic flux and the limited availability of the universal isoprenoid precursors complicate their widespread implementation. The artificial pathways developed thus far have been solely focused on synthesizing dimethylallyl and isopentenyl diphosphates and require additional enzymes for the generation of polyprenyl-diphosphates (polyprenyl-PPs). Thus, the primary objective of this proposal is to develop an efficient strategy for the synthesis of both natural and unnatural (poly)prenyl-PPs for downstream applications. This will be achieved using two complementary methods: i) employing undecaprenol kinases and isopentenyl phosphate kinases; and ii) employing hydroxyethylthiazole kinase, isopentenyl phosphate kinases, and farnesyl diphosphate synthase. Additionally, the two methods will work in conjunction with isoprenoid methylatransferases to incorporate additional diversity into the polyprenyl- PPs. The proposed studies include: i) structural and functional assessment of selected enzymes, ii) catalyst engineering, and iii) optimization of coupled in vitro and in vivo platforms for the generation of diversified libraries of select natural products. We expect these studies to generate: i) rules and concepts to advance knowledge on structure-activity relationships in selected classes of enzymes; ii) an optimized, enzyme-coupled platform to generate diversified substrates and isoprenoids; and iii) novel isoprenoid analogs with potential therapeutic applications. Thus, the proposed work will offer unprecedented access to uniquely bioactive isoprenoid libraries not readily accessible via traditional methods, and it stands to deepen our fundamental understanding of four enzyme classes while also developing them into useful biocatalysts.

资助的父母奖的摘要（1R01GM138800-01A1）。类异型代表多种类别 具有广泛应用在医学和工业中的化合物。它们从自然来源中提取的是 具有挑战性和潜在对环境有害，而许多人的巨大结构复杂性 类异型使传统的化学合成非平凡。现代代谢工程和合成生物学 方法已经克服了其中一些困难，但是与代谢通量有关的问题和有限的问题 普遍的类异戊二烯前体的可用性使它们的广泛实现变得复杂。人造 到目前为止，开发的途径仅集中在合成二甲基二甲基二磷酸二甲基二磷酸酯上 并需要额外的酶来产生聚丙烯基 - 二磷酸盐（聚丙烯基-PPS）。因此， 该提案的主要目的是制定一种有效的策略来综合自然和不自然的 （Poly）下游应用的丙烯酰PPP。这将使用两种互补方法来实现：i） 采用异孕醇激酶和等烯基磷酸激酶； ii）采用羟基乙醇 激酶，异戊烯基磷酸激酶和Farnesyl二磷酸合酶。另外，这两种方法将 与异澳甲基转移酶一起工作，将额外的多样性纳入聚丙烯基 - pps。拟议的研究包括：i）选定酶的结构和功能评估，ii）催化剂 工程和iii）优化在体外和体内平台，以生成多样化 精选天然产品的库。我们希望这些研究能够产生：i）规则和概念可以提高 关于选定酶类别的结构活性关系的知识； ii）优化的酶耦合 产生多样化的底物和类异丙因的平台； iii）具有潜力的新型类似物类似物 治疗应用。因此，拟议的工作将提供前所未有的访问权限 类异丙库不容易通过传统方法访问，它可以加深我们的基本 了解四个酶类，同时也将它们发展为有用的生物催化剂。