Engineering Dynamic Control of Natural Product Biosynthesis in Bacteria

细菌天然产物生物合成的工程动态控制

• 批准号: 10469475
• 负责人: Yajun Yan
• 金额: $ 35.09万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10469475
• 关键词: 5-Hydroxytryptophan; Acids; Address; Anabolism; Antisense RNA; Bacteria; Biological; Biosensor; Cells; Decision Making; Development; Down-Regulation; Engineering; Escherichia coli; Flavanones; Flavonoids; Gene Expression; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic; Growth; Indole Alkaloids; Intelligence; Lead; Logic; Metabolic; Metabolism; Natural Products; Output; Pharmacologic Substance; Phenotype; Plants; Production; Property; Protein Engineering; Regulation; Research; Research Activity; Salicylic Acids; Techniques; Terpenes; Therapeutic Effect; Tryptophan; Up-Regulation; base; biological systems; environmental change; gene repression; improved; insight; large scale production; metabolic engineering; microbial; naringenin; novel; promoter; real world application; response; synthetic biology; tool

Project Summary Engineering natural product biosynthesis in bacteria represents a promising strategy to produce these high-value and pharmaceutically important compounds. However, to achieve economically viable production is quite difficult and challenging. To address this challenge, establishing artificial logic and dynamic control functions in cells by mimicking natural regulations in biological systems has been developed and becomes a powerful approach to enable superior microbial production. This approach render great biological robustness to the cells and allows them to autonomously adjust their metabolic states and activities by sensing the cellular and environmental changes. However, compared with natural regulations that exert simultaneous and orthogonal up- and down- regulation on various gene targets, the current dynamic control techniques mainly relies on simple gene circuits to perform mono-function, either up- or down-regulation, on a limited set of genes, which are still rudimentary and less sophisticated. To bridge these gaps, development of new genetic tools and their use in novel control strategies are highly desired. Recently, the PI’s lab has achieved the biosynthesis of natural products with defined pharmaceutical properties and therapeutic effects such as 4-hydroxycoumarin and 5-hydroxytryptophan in E. coli through synthetic biology, metabolic engineering and protein engineering approaches and developed antisense RNA tools for intervening cellular metabolism to enhance the biosynthesis of plant polyketides, such as flavanone naringenin. In this MIRA proposal, we aim at exploring a new and general strategy for developing more sophisticated dynamic control network to greatly enhance natural product biosynthesis in bacteria. The dynamic control network is expected to have the capability of implementing autonomous and intelligent up- regulation and down-regulation functions in response to cellular and environmental changes and maintain the cells at optimal production states throughout all growth stages to achieve maximal production efficiency. Our recent progress on the biosynthesis of 4-hydroxycoumarin, 5-hydroxytryptophan and the development of antisense RNAs tools will serve as the groundwork for us to understand how to efficiently establish and implement the dynamic control network and decision-making strategies in cells for real-world applications. Specifically, four coherent projects with distinct research activities will be pursued, which include: 1) developing salicylic acid, p-coumaric acid and tryptophan responsive promoters for up-regulation of gene expression; 2) expanding antisense RNA tools for controllable down-regulation of gene expression; 3) developing and characterizing promoter and antisense RNA based dynamic control network; 4) applying dynamic control network to improve biosynthesis of 4-hydroxycoumarin, flavanone naringenin and 5-hydroxytryptophan in E. coli.

项目摘要 细菌中的工程天然产品生物合成代表了产生这些高价值的希望策略 和物理上重要的化合物。但是，要实现经济可行的生产非常困难 和挑战。为了应对这一挑战，通过通过 模仿生物系统中的自然法规已经开发出来，并成为一种有力的方法 实现优质微生物产生。这种方法为细胞提供了极大的生物鲁棒性，并允许 他们通过感测细胞和环境来自主调整其代谢状态和活动 更改。但是，与对上下简单和正交的自然法规相比 对各种基因靶标的调节，当前的动态控制技术主要依赖于简单的基因电路 在有限的基因上执行上调或下调的单功能，这些基因仍然是基本的 而且不那么精致。弥合这些差距，开发新的遗传工具及其在新颖控制中的使用 策略是高度期望的。最近，Pi的实验室实现了天然产品的生物合成 定义的药物特性和治疗作用，例如4-羟基丙木蛋白和5-羟基化的效果 在大肠杆菌中通过合成生物学，代谢工程和蛋白质工程方法开发 反义RNA工具用于干预细胞代谢以增强植物聚酮化合物的生物合成，例如 作为黄酮Naringenin。在此Mira提案中，我们旨在探索一种新的一般策略 更复杂的动态控制网络可大大增强细菌中天然产物生物合成。 预计动态控制网络具有实施自主和智能上升的能力 调节和下调功能响应细胞和环境变化，并维持 在所有生长阶段，最佳生产状态的细胞以达到最大的生产效率。我们的 4-羟基木菊，5-羟基tryptypophan的生物合成的最新进展和 反义RNA工具将成为我们了解如何有效建立和 在现实世界应用中，在单元格中实施动态控制网络和决策策略。 具体而言，将进行四个具有不同研究活动的连贯项目，其中包括：1）开发 水杨酸，p-墨酸和色氨酸反应启动子，用于上调基因表达； 2） 扩展反义RNA工具，以控制基因表达的下调； 3）发展和 表征启动子和基于反义RNA的动态控制网络； 4）应用动态控制网络 为了改善大肠杆菌中4-羟基丙烯蛋白酶，黄酮烯酸酯和5-羟基tryptypophan的生物合成。