Design of a bacterial consortium for consolidated bioprocessing

主要研究者：Schmidt-Dannert，Claudia提案编号：1235714机构：明尼苏达大学双城分校标题：用于综合生物加工的细菌财团的设计生物技术生物过程通常使用代谢工程微生物菌株的纯培养物。相比之下，环境过程通常利用天然微生物聚生体中存在的代谢多样性来完成单个生物体无法单独完成的任务。寻求开发用于生物燃料生产的整合生物工艺，其中将柠檬酸木质纤维素生物质的水解和随后的糖转化为生物燃料相结合，已经重振了对混合培养物的使用的研究。目前正在寻求不同的策略，以获得菌株，都是木质纤维素和先进的生物燃料生产商，但到目前为止，没有一个工程菌株是有效的在这两个tasks.The工程的生物网络与新出现的属性在合成生物学领域的应用提供了工具和策略，它成为可能的设计和研究合成微生物财团。通过将代谢功能区室化到单独的微生物种群中，每个菌株的代谢网络可以针对共同导致产生所需化合物的不同任务进行优化。因此，在生物过程中共培养的不同工程化微生物菌株可以被认为是单独的代谢模块，其可以与另外的模块交换或增强。然而，这种混合微生物群落工程的先决条件是建立种群控制系统。本项目将设计一个原型合成微生物群落，作为综合生物加工的概念验证。从拟议的研究结果可以指导其他生物过程的开发使用工程微生物财团。具体来说，本项目将设计一个由B组成的混合文化。subtilis、不透明红球菌（Rhodococcus opacus）和E.大肠杆菌进行生物柴油的从头生产。工业蛋白表达宿主B。枯草芽孢杆菌将被改造为分泌纤维素分解酶，用于从生物质中释放糖，而产油R. opacus和产乙醇E.大肠杆菌菌株一起将糖转化为脂肪酸乙酯（生物柴油）。开发这样一个联合体的关键是设计和配置适当的系统，以根据工艺要求控制每个联合体成员的生长。该项目将利用细菌群体感应系统的组成部分设计这种种群控制系统。本研究将建立一个系统模拟模型，以指导我们设计这些控制系统。本项目将为工程微生物菌群在生物加工中的开发和应用提供概念验证。因此，混合培养物在生物技术中的应用可能会重新焕发活力，并从前DNA技术转移到合成生物学领域。从这个应用程序的结果将提供的见解和模型的人口控制系统的设计需要调整人口规模的要求，一个特定的过程。拟议的研究将影响生物技术和合成生物学领域。这项研究的结果可能会为生物燃料生产和其他生物生产过程带来新的和更有效的方法。 拟议的研究将为一名博士后研究人员和四名本科生提供遗传和代谢工程方面的培训。

PI: Schmidt-Dannert, ClaudiaProposal Number: 1235714Institution: University of Minnesota-Twin CitiesTitle: Design of a bacterial consortium for consolidated bioprocessingBiotechnological bioprocesses typically use pure cultures of metabolically engineered microbial strains. In contrast, environmental processes generally utilize the metabolic diversity present in native microbial consortia to accomplish tasks that not one single organism can do alone. The quest for the development of consolidated bioprocesses for biofuel production, where hydrolysis of recalcitrant lignocellulosic biomass and subsequent sugar conversion into biofuels are combined, has reinvigorated investigations into the use of mixed cultures. Currently different strategies are being pursued to obtain strains that are both lignocellulolytic and advanced biofuel producers, but so far none of the engineered strains is efficient at both tasks.The engineering of biological networks with new emergent properties employed in the field of synthetic biology has provided tools and strategies with which it becomes possible to design and investigate synthetic microbial consortia. By compartmentalizing metabolic functions into separate microbial populations, the metabolic network of each strain can be optimized for distinct tasks that together result in the production of a desired compound. The different engineered microbial strains co-cultured in a bioprocess can therefore be considered as individual metabolic modules that may be exchanged or augmented with additional modules. Prerequisite for the engineering of such mixed microbial communities, however, is the establishment of population control systems.This project will design a prototype synthetic microbial community as a proof-of-concept for consolidated bioprocessing. Results from the proposed research can guide the development of other bioprocesses using engineered microbial consortia. Specifically, this project will engineer a mixed culture consisting of B. subtilis, Rhodococcus opacus and E. coli for de novo biodiesel production. The industrial protein expression host B. subtilis will be engineered to secrete cellulolytic enzymes for sugar release from biomass, while the oleaginous R. opacus and the ethanologenic E. coli strain together will convert sugars into fatty acid ethyl esters (biodiesel). Key to the development of such a consortium is the engineering and configuration of adequate systems to control growth of each consortium member in accordance to the requirements of the process. The project will design such population control systems using components of bacterial quorum sensing systems. A system simulation model will be built to guide us in the design these control systems.The project will provide a proof-of-concept for the development and application of engineered microbial consortia for bioprocessing. The use of mixed cultures for biotechnology may thus be revitalized and moved from the pre-DNA technology into the synthetic biology area. Results from this application will provide insights and a model for the design of population control systems needed to adjust population sizes to the requirements of a specific process. The proposed research will impact the field of biotechnology and synthetic biology. Findings of this research may lead to new and more efficient approaches for biofuel production and other bioproduction processes. The proposed research will provide training in genetic and metabolic engineering for one postdoctoral researcher and four undergraduate students.