Enzyme-Mediated Synthesis of Functionalized Terpene Structures

酶介导的功能化萜烯结构的合成

• 批准号: 7819500
• 负责人: JAY D KEASLING
• 金额: $ 50万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7819500
• 关键词: Address; Alkenes; Anabolism; Anti-Bacterial Agents; Antifungal Agents; Antineoplastic Agents; Area; Bacillus megaterium; Biological Factors; Characteristics; Chemistry; Complementary DNA; Complex; Cytochrome P450; Data; Development; Diterpenes; Engineering; Environment; Enzymes; Euphorbiaceae; Family; Genes; Health; Human; Incidence; Laboratories; Mediating; Medical; Metabolic Biotransformation; Metals; Methods; Mixed Function Oxygenases; Mutagenesis; Organic Synthesis; Pharmaceutical Preparations; Plants; Production; Protein Engineering; Reaction; Route; Sesquiterpenes; Solvents; Source; Structure; Structure-Activity Relationship; Technology; Terpenes; Testing; Therapeutic; Work; abstracting; analog; base; biological systems; catalyst; chemical synthesis; commercialization; cost; design; drug candidate; drug development; drug discovery; functional group; human disease; improved; in vivo; interest; member; microbial; microbial host; mutant; novel; novel therapeutics; oxidation; programs; public health relevance; terpene synthase; therapeutic target; tool

DESCRIPTION (provided by applicant): Enzyme-Mediated Synthesis of Functionalized Terpene Structures Project Summary/Abstract This application addresses broad challenge area (06), Enabling Technologies, and specific Challenge Topic 06-GM-109: Green chemistry and engineering for drug discovery, development, and production. Terpenes, constituting one of the most diverse groups of compounds synthesized by biological systems, have been used as antibacterial, antifungal, and anticancer agents in the treatment of human disease. Despite the high value of many terpenes as therapeutic targets, chemical synthesis routes have proven elusive due to the presence of several chiral centers in the terpene core and the need for differential protection and deprotection of functional groups. In the last ten years there has been tremendous progress in the chemical synthesis of complex functionalized terpenes using metal-based catalysts to execute regio and stereospecific reactions, and having access to small quantities of terpenes has been sufficient to confirm the therapeutic value of a number of drug candidates. However, going forward we will need cost-effective and green synthetic methods to produce quantities of therapeutic terpenes sufficient for drug development and commercialization. To address this problem, we propose a green chemistry platform for the synthesis of terpene olefins and functionalized terpenes of therapeutic value. First, through expression of selected known and novel terpene synthases in engineered microbial hosts, we will generate terpene olefins that constitute core structures of therapeutic compounds. Second, we will functionalize these terpene olefins using mutants of a highly efficient bacterial cytochrome P450 monooxygenase. Finally, we will optimize expression of these enzymes in microbial strains that we have engineered to overproduce universal terpene precursors in order to achieve high-level production of terpene olefins and oxidized terpenes of therapeutic value. Successful completion of this proposal will provide a microbial platform for the economical and clean synthesis of terpene olefins and oxidized terpenes of therapeutic value. This work has three significant impacts on human health. First, the microbial terpenes can then be used as late intermediates in the synthesis of complete therapeutic terpenes and terpene analogs, thus accelerating the discovery of terpene-based therapeutics. Second, microbial production of terpenes is more cost-effective and environment-friendly when compared to traditional organic synthesis methods that rely on expensive starting materials and toxic solvents and catalysts. Finally, the generality of the proposed platform will streamline the synthesis of other functionalized terpenes via the introduction and mutagenesis of new enzymes in our host strains. PUBLIC HEALTH RELEVANCE: There are many natural products in the terpene family with promising or proven therapeutic characteristics. As yields from natural sources are frequently low, and typical chemical synthesis routes are often challenging and expensive, we aim to facilitate economical and clean enzyme-mediated synthesis of a number of these products. In addition, our plan enables green synthesis of product analogs for drug discovery and improvement programs though the use of mutant enzymes.

描述（由申请人提供）：官能化萜烯结构的酶介导合成项目概述/摘要本申请涉及广泛的挑战领域（06），使能技术，和特定的挑战主题06-GM-109：用于药物发现、开发和生产的绿色化学和工程。萜类化合物是生物系统合成的种类最多的化合物之一，在人类疾病的治疗中已被用作抗细菌剂、抗真菌剂和抗癌剂。尽管许多萜烯作为治疗靶点具有很高的价值，但由于萜烯核心中存在几个手性中心以及需要对官能团进行差异保护和脱保护，化学合成途径已被证明是难以捉摸的。在过去的十年中，在使用基于金属的催化剂来执行区域和立体特异性反应的复杂官能化萜烯的化学合成方面已经取得了巨大的进展，并且能够获得少量萜烯已经足以证实许多候选药物的治疗价值。然而，展望未来，我们将需要成本效益和绿色合成方法来生产足够用于药物开发和商业化的治疗性萜烯。为了解决这个问题，我们提出了一个合成具有治疗价值的萜烯烯烃和功能化萜烯的绿色化学平台。首先，通过在工程化微生物宿主中表达选定的已知和新的萜烯脱氢酶，我们将产生构成治疗性化合物的核心结构的萜烯烯烃。其次，我们将功能化这些萜烯烯烃使用突变体的高效细菌细胞色素P450单加氧酶。最后，我们将优化这些酶在微生物菌株中的表达，我们已经设计了过量生产通用萜烯前体，以实现高水平生产萜烯烯烃和具有治疗价值的氧化萜烯。该提案的成功完成将为具有治疗价值的萜烯烯烃和氧化萜烯的经济和清洁合成提供微生物平台。这项工作对人类健康有三大影响。首先，微生物萜烯可用作合成完全治疗性萜烯和萜烯类似物的后期中间体，从而加速基于萜烯的治疗剂的发现。其次，与依赖昂贵的起始材料和有毒溶剂和催化剂的传统有机合成方法相比，萜烯的微生物生产更具成本效益和环境友好性。最后，所提出的平台的通用性将通过在我们的宿主菌株中引入和诱变新酶来简化其他功能化萜烯的合成。 公共卫生相关性：在萜烯家族中有许多天然产品具有有前途或已证实的治疗特性。由于天然来源的产率通常较低，并且典型的化学合成路线通常具有挑战性且昂贵，因此我们的目标是促进许多这些产品的经济和清洁的酶介导合成。此外，我们的计划使绿色合成的产品类似物的药物发现和改进计划，通过使用突变酶。