Engineering microbial co-cultures for complex natural product biosynthesis

用于复杂天然产物生物合成的工程微生物共培养

• 批准号: 1706058
• 负责人: Haoran Zhang
• 金额: $ 29.91万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1706058&HistoricalAwards=false
• 关键词: Engineering; microbial; co; cultures; complex

Many naturally-occurring pharmaceutical molecules (for example: antibiotics, anti-cancer drugs) are the result of a long sequence of reactions that add, piece by piece, different subunits to the growing molecule, in a fashion similar to an industrial assembly line. When production occurs in a single organism, it places a large burden on the organism to provide all of the raw materials and energy needed to complete construction. This project will investigate a strategy that creates organisms capable of efficiently producing a particular subunit of the larger drug molecule, "subcontracts" production of subunits to multiple organisms, then brings the subunits together to create the final product. If successful, this could dramatically improve the efficiency of large molecule drug production, and could also increase the speed of identification of more effective drug molecules. This project will also expand the development of a STEM-trained workforce through targeted outreach and engagement of high school and college students from under-represented populations in the research project.This proposed study aims to adapt engineered E. coli strains to constitute co-cultures for production of three representative natural products with characteristically long and complicated biosynthetic pathways. Each biosynthetic pathway is divided into two independent modules, each of which is accommodated in a different, specialized E. coli strain. Such designed pathway modularization allows for reduction of metabolic burden and improvement of biosynthetic performance of the co-culture strains, as each strain is only responsible for one portion of the biosynthesis. The involved strains will also be metabolically engineered individually to meet the needs of the designated pathway enzymes and maximize their bioconversion capabilities. Moreover, the biosynthetic strength of the individual pathway modules will be balanced for bioproduction optimization through manipulation of the relative ratio of the co-culture populations. The microbial systems will be scaled up to investigate population stability and bioproduction behavior under high cell density conditions. This project offers outstanding potential to design and engineer the production of complex molecules. The fundamental knowledge gained regarding the dynamic regulation of population ratios and metabolic activity in mixed populations will provide insight into the behavior of natural microbial mixed populations, which will have implications for a wide variety of industrial and environmentally-important processes.

许多天然存在的药物分子（例如：抗生素、抗癌药物）是一长串反应的结果，这些反应以类似于工业装配线的方式将不同的亚基逐个添加到不断生长的分子中。当生产发生在单一有机体中时，它会给该有机体带来很大的负担，以提供完成建设所需的所有原材料和能源。该项目将研究一种策略，该策略创建能够有效生产较大药物分子的特定亚基的生物体，将亚基的生产“分包”给多个生物体，然后将亚基组合在一起以创建最终产品。如果成功，这可以显着提高大分子药物生产的效率，并且还可以提高识别更有效药物分子的速度。该项目还将通过有针对性地推广和吸引来自代表性不足人群的高中生和大学生参与研究项目，扩大受过 STEM 培训的劳动力队伍的发展。这项拟议的研究旨在改造工程大肠杆菌菌株以构成共培养物用于生产三种具有代表性的天然产物，这些天然产物具有漫长而复杂的生物合成途径。每个生物合成途径分为两个独立的模块，每个模块都容纳在不同的、专门的大肠杆菌菌株中。这种设计的途径模块化可以减少共培养菌株的代谢负担并提高生物合成性能，因为每种菌株仅负责生物合成的一部分。所涉及的菌株也将单独进行代谢工程，以满足指定途径酶的需求并最大限度地提高其生物转化能力。此外，通过操纵共培养群体的相对比例，可以平衡各个途径模块的生物合成强度，以实现生物生产优化。微生物系统将扩大规模，以研究高细胞密度条件下的群体稳定性和生物生产行为。该项目为复杂分子的设计和工程生产提供了巨大的潜力。获得的关于混合种群中种群比例和代谢活动的动态调节的基础知识将有助于深入了解自然微生物混合种群的行为，这将对各种工业和环境重要的过程产生影响。

项目成果

Establishing microbial co‐cultures for 3‐hydroxybenzoic acid biosynthesis on glycerol

建立用于甘油生物合成 3-羟基苯甲酸的微生物共培养物

• DOI: 10.1002/elsc.201800195
• 发表时间: 2019-05-01
• 期刊: Engineering in Life Sciences
• 影响因子: 2.7
• 作者: Yiyao Zhou;Zhenghong Li;Xiaonan Wang;Haoran Zhang
• 通讯作者: Haoran Zhang

Constructing E. coli Co‐Cultures for De Novo Biosynthesis of Natural Product Acacetin

构建用于天然产物金合欢素从头生物合成的大肠杆菌培养物

• DOI: 10.1002/biot.202000131
• 发表时间: 2020-06-23
• 期刊: Biotechnology Journal
• 影响因子: 4.7
• 作者: Xiaonan Wang;Alan Shao;Zhenghong Li;Lizelle Policarpio;Haoran Zhang
• 通讯作者: Haoran Zhang

Balancing the non-linear rosmarinic acid biosynthetic pathway by modular co-culture engineering

通过模块化共培养工程平衡非线性迷迭香酸生物合成途径

• DOI: 10.1016/j.ymben.2019.03.002
• 发表时间: 2019-07
• 期刊: Metabolic Engineering
• 影响因子: 8.4
• 作者: Li, Zhenghong;Wang, Xiaonan;Zhang, Haoran
• 通讯作者: Zhang, Haoran

De novo biosynthesis of complex natural product sakuranetin using modular co-culture engineering

使用模块化共培养工程从头生物合成复杂天然产物樱花素

• DOI: 10.1007/s00253-020-10576-1
• 发表时间: 2020-04-13
• 期刊: Applied Microbiology and Biotechnology
• 影响因子: 5
• 作者: Xiaonan Wang;Zhenghong Li;Lizelle Policarpio;M. Koffas;Haoran Zhang
• 通讯作者: Haoran Zhang

Utilizing cross-species co-cultures for discovery of novel natural products

利用跨物种共培养来发现新型天然产物

• DOI: 10.1016/j.copbio.2021.01.023
• 发表时间: 2021-06
• 期刊: Current Opinion in Biotechnology
• 影响因子: 7.7
• 作者: Zhuang, Lei;Zhang, Haoran
• 通讯作者: Zhang, Haoran