UNS: Engineering Stable Two- and Three-Component Bacterial Consortia

自然产生的微生物群落是共同完成复杂任务的微生物群落。该项目旨在利用一系列具有良好特征的遗传相互作用来产生稳定、可调的微生物群落。这一想法的应用将在生物可再生能源生产的两个方面得到证明。在第一个演示中，将讨论混合糖的同时利用问题。木质纤维素生物质含有多种类型的糖，将这些糖转化为生物可再生燃料和化学品将最佳地利用所有这些糖。将建立一个能够利用三种最常见的生物质衍生糖的微生物联合体。第二个演示涉及一个联合体，其中所有成员消耗相同的糖（葡萄糖），但生产两种不同的产品。这项工作将开发新技术，使生物可再生燃料和化学品的生产得到改善。该项目将为本科生和研究生的研究人员提供培训，并为K12的外展活动提供基础。本提案的目标是重新设计一个现有的产生联合体的大肠杆菌遗传回路，以形成微生物联合体，利用混合底物和生产混合产品。这个联盟将不同于现有的联盟，因为它将是(i)等基因和（ii）自我调整。虽然这个联合体以离散的表达状态存在，但其成员在基因上都是相同的，并且不需要为了维持稳定的种群而对营养缺陷特征进行工程改造。自调谐行为将确保种群分布将响应外部或代谢信号而发生变化。这种等基因群体获得这种离散和自我调节行为的能力是基于在发病机制中pap和膜操纵子DNA复制后不久发生的一组复杂的相互作用。这项工作涉及生物分子过程的基础工程研究，可能导致生物技术和生物能源工业的使能技术的发展。具体来说，将展示生物质衍生糖的利用和生物可再生化学品的生产。这项工作的具体目标包括(i)报告系统的构建和在各种生长条件下的等基因、自调谐联合体的表征；（ii）同时自我调节葡萄糖、木糖和阿拉伯糖消耗的系统工程；（iii）联合生产琥珀酸甲酯和1,4-丁二醇（BDO）系统的工程设计。该奖项由CBET部门的生物技术和生化工程项目颁发，由分子和细胞生物学部门的系统和合成生物学项目共同资助。

1511646 Jarboe, Laura Naturally-occurring microbial consortia are communities of microorganisms that work together to perform complex tasks. This project aims to use a well-characterized series of genetic interactions to generate stable, tunable microbial consortia. The application of this idea will be demonstrated in two aspects of the production of biorenewables. In the first demonstration, the problem of simultaneous utilization of mixed sugars will be developed. Lignocellulosic biomass contains a variety of sugar types and conversion of these sugars to biorenewable fuels and chemicals would optimally utilize all of these sugars. A microbial consortium capable of utilizing the three most common biomass-derived sugars will be established. The second demonstration involves a consortium in which all members consume the same sugar (glucose), but produce two distinct products. This work will develop new technology that will enable improved production of biorenewable fuels and chemicals. This project will provide training for undergraduate and graduate researchers and provide a basis for K12 outreach activities. The goal of this proposal is to redesign an existing consortium-generating Escherichia coli genetic circuit to form microbial consortia for utilization of mixed substrates and production of mixed products. This consortium will be distinct from existing consortia in that it will be (i) isogenic and (ii) self-tuning. While the consortium exists in discrete expression states, the members are all genetically identical and will not require the engineering of auxotrophic features in order to maintain a stable population. The self-tuning behavior will ensure that the population distribution will shift in response to external or metabolic signals. The ability of this isogenic population to attain such discrete and self-tuning behavior is based on a complex set of interactions that occur shortly after DNA replication in the pap and fim operons in the context of pathogenesis. This work involves fundamental engineering research of biomolecular processes that could lead to the development of enabling technology for biotechnology and bioenergy industries. Specifically, the utilization of biomass-derived sugars and the production of biorenewable chemicals will be demonstrated. The specific aims of this work include (i) the construction of reporter systems and characterization of the isogenic, self-tuning consortium in a variety of growth conditions; (ii) the engineering of the system for simultaneous, self-tuning consumption of glucose, xylose and arabinose; (iii) the engineering of the system for co-production of methylsuccinate and 1,4-butanediol (BDO).This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology.