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

基本信息

批准号：
10543814
负责人：
Shanteri Singh
金额：
$ 29.11万
依托单位：
UNIVERSITY OF OKLAHOMA
依托单位国家：
美国
项目类别：
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/10543814
关键词：
Acids Active Sites Address Alcohols Cannabinoids Cells Collaborations Coupled Coupling Diphosphates Engineering Environment Enzymes Escherichia coli Farnesol 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 Natural Selections Natural Source Nature North Carolina Pathway interactions Phosphorylation Phosphotransferases Process Production Reaction S-Adenosylhomocysteine S-Adenosylmethionine Squalene Squalene Synthetase Structure Structure-Activity Relationship System Technology Therapeutic Universities Variant Work analog analytical method 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

Acids Active Sites Address Alcohols Cannabinoids Cells Collaborations Coupled Coupling Diphosphates Engineering Environment Enzymes Escherichia coli Farnesol 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 Natural Selections Natural Source Nature North Carolina Pathway interactions Phosphorylation Phosphotransferases Process Production Reaction S-Adenosylhomocysteine S-Adenosylmethionine Squalene Squalene Synthetase Structure Structure-Activity Relationship System Technology Therapeutic Universities Variant Work analog analytical method 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

展开

项目摘要

Project Summary 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.

项目摘要类异型代表着各种各样的化合物，在医学和工业中使用了广泛的应用。它们从自然来源中提取既有挑战性又可能对环境有害，而许多类异戊二烯的巨大结构复杂性使传统的化学合成无繁殖。现代的代谢工程和合成生物学方法已经克服了其中一些困难，但问题与代谢通量和普遍类异丙类动物前体的有限可用性有关使其复杂性有关广泛的实现。迄今为止开发的人工途径仅专注于合成二甲基烯丙基和异戊烯基二磷酸盐，需要额外的酶以产生聚丙烯基 - 双磷酸盐（聚丙烯基-PP）。因此，该提案的主要目的是制定有效的策略在下游应用中综合自然和非自然（多年）前PPP。这将被实现使用两种互补方法：i）采用异孕醇激酶和异戊烯基磷酸激酶；和 ii）采用羟基硫唑激酶，异戊烯基磷酸激酶和Farnesyl二磷酸合酶。此外，这两种方法将与异澳甲基转移酶结合起来合并聚丙基培养基的额外多样性。拟议的研究包括：i）结构和功能评估选定的酶，ii）催化剂工程和iii）优化体外和体内平台的耦合精选天然产品的多元化库产生。我们希望这些研究能够产生：i）规则和促进选定酶类中结构活性关系知识的概念； ii）优化，酶耦合平台，以产生多样化的底物和类异丙裔； iii）新型类似物类似物具有潜在的治疗应用。因此，拟议的工作将提供前所未有的访问权限生物活性类异丙库不容易通过传统方法访问，它可以加深我们对四种酶类别的基本了解，同时也将它们发展为有用的生物催化剂。