Enzyme-Mediated Synthesis of Functionalized Terpene Structures

酶介导的功能化萜烯结构的合成

基本信息

批准号：
7940803
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
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7940803
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

项目摘要

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）：酶介导的功能化萜烯结构的合成项目摘要/摘要该申请涉及广泛的挑战领域（06），促成技术和特定的挑战主题06-gm-109：用于药物发现，开发，开发和生产的绿色化学和工程。萜烯构成由生物系统合成的最多样化的化合物之一，已被用作抗菌，抗真菌和抗癌药物治疗人类疾病的抗菌药物。尽管许多萜烯作为治疗靶标具有很高的价值，但由于萜烯核心中存在几个手性中心，并且需要对官能团的差异保护和脱旋触摸，因此被证明是难以捉摸的。在过去的十年中，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取得了巨大进展，并且可以访问少量的萜烯，足以确认许多药物候选者的治疗值。但是，未来，我们将需要具有成本效益和绿色合成方法，以产生足以用于药物开发和商业化的治疗萜烯。为了解决这个问题，我们提出了一个绿色化学平台，用于合成萜烯烯烃和治疗价值的能键烯。首先，通过在工程微生物宿主中的某些已知和新型萜烯合酶的表达，我们将生成构成治疗化合物核心结构的萜烯烯烃。其次，我们将使用高效的细菌细胞色素P450单加氧酶的突变体功能化这些萜烯烯烃。最后，我们将优化这些酶在微生物菌株中的表达，我们已经设计为过量产生通用萜烯前体，以实现萜烯烯烃的高水平产生和治疗价值的氧化萜烯。该提案的成功完成将提供一个微生物平台，用于经济和清洁的萜烯烯烃和治疗价值的氧化萜烯。这项工作对人类健康有三个重大影响。首先，将微生物萜烯用作完整的治疗萜和萜类似物的合成中的晚期中间体，从而加速了基于萜烯的治疗剂的发现。其次，与传统的有机合成方法相比，萜烯的微生物生产更具成本效益和环境友好，这些方法依靠昂贵的起始材料，有毒的溶剂和催化剂。最后，所提出的平台的一般性将通过我们的宿主菌株中新酶的引入和诱变来简化其他功能化萜烯的合成。公共卫生相关性：萜烯家族中有许多天然产品，具有有希望或经过证实的治疗特征。由于自然来源的产量通常很低，而且典型的化学合成途径通常具有挑战性且昂贵，因此我们旨在促进经济和清洁酶介导的许多这些产品的合成。此外，我们的计划可以通过使用突变酶来绿色合成用于药物发现和改进程序的产品类似物。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Functional characterization of four sesquiterpene synthases from Ricinus communis (castor bean).

DOI：
10.1016/j.phytochem.2012.02.022
发表时间：
2012-06
期刊：
Phytochemistry
影响因子：
3.8
作者：
Xinkai Xie;J. Kirby;J. Keasling
通讯作者：
Xinkai Xie;J. Kirby;J. Keasling

Versatile synthesis of probes for high-throughput enzyme activity screening.

用于高通量酶活性筛选的探针的多功能合成。

DOI：
10.1007/s00216-013-6888-z
发表时间：
2013
期刊：
Analytical and bioanalytical chemistry
影响因子：
4.3
作者：
deRond,Tristan;Peralta-Yahya,Pamela;Cheng,Xiaoliang;Northen,TrentR;Keasling,JayD
通讯作者：
Keasling,JayD