Collaborative Research: A Systems Biology Approach for Metabolically Engineering Escherichia coli for Producing Hydrogen via Fermentation

合作研究：通过代谢工程大肠杆菌发酵生产氢气的系统生物学方法

• 批准号: 1212320
• 负责人: Thomas Wood
• 金额: $ 5.99万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1212320&HistoricalAwards=false
• 关键词: Collaborative; Research; Systems; Biology; Approach

CBET-0753702WoodNon-harmful bacteria may be used to produce chemicals for humans and this includes fuels. Molecular hydrogen is an environmentally-clean fuel; hence, if hydrogen can be synthesized efficiently by bacteria, then we may reduce air pollution, reduce our dependence on foreign oil, and avoid greenhouse gases (the carbon dioxide formed by our process will be recycled to plant mass and used again to form hydrogen so there are no net emissions). This research will enable hydrogen to be produced as a clean form of energy from bacteria using fermentation of simple sugars. In fermentative production of hydrogen, glucose is converted into pyruvate which is converted to formate which is then converted to hydrogen. Hence, it is imperative to direct the carbon flux in the bacterial cell to hydrogen formation by preventing competing glucose reactions and those that convert pyruvate to acetate, lactate, and ethanol or that remove formate. To achieve this aim, a systems biology approach will be used to produce more hydrogen that includes (i) altering the metabolism of the bacteria by removing unwanted pathways, (ii) evolving the necessary enzymes using protein engineering, and (iii) modeling all the metabolic pathways of the bacterial cell so that even better bacteria may be generated. We have chosen Escherichia coli as our model system since (i) all 3985 non-lethal single mutations of strain E. coli BW25113 are available; (ii) we may combine mutations rapidly using our novel, successive, virus-based method; (iii) E. coli is the best-studied bacterium so its metabolism is well understood; and (iv) its genome is sequenced so DNA microarrays are available and metabolic flux analysis may be carried out at the genome-scale. Along with the research results, the three professors involved in this research will train undergraduate and graduate students, will make accessible, via the World-Wide Web, specific outcomes of the project, and will disseminate some of the research results to a diverse audience using podcasting.

CBET-0753702木材无害细菌可用于为人类生产化学品，其中包括燃料。 分子氢是一种环保清洁燃料；因此，如果细菌可以有效地合成氢气，那么我们就可以减少空气污染，减少对外国石油的依赖，并避免温室气体排放（我们的过程中形成的二氧化碳将被回收到植物体中并再次用于形成氢气，因此没有净排放）。 这项研究将使细菌能够利用单糖的发酵来生产氢气作为一种清洁能源。 在氢的发酵生产中，葡萄糖转化为丙酮酸，丙酮酸转化为甲酸盐，然后甲酸盐转化为氢。 因此，必须通过防止竞争性葡萄糖反应以及将丙酮酸转化为乙酸盐、乳酸和乙醇或去除甲酸盐的反应，将细菌细胞中的碳通量引导至氢形成。 为了实现这一目标，将使用系统生物学方法来生产更多的氢气，包括（i）通过消除不需要的途径来改变细菌的新陈代谢，（ii）使用蛋白质工程进化必要的酶，以及（iii）对所有氢进行建模细菌细胞的代谢途径，以便可以产生更好的细菌。 我们选择大肠杆菌作为我们的模型系统，因为 (i) 大肠杆菌 BW25113 菌株的所有 3985 个非致死单突变均可用； (ii) 我们可以使用我们新颖的、连续的、基于病毒的方法快速组合突变； (iii) 大肠杆菌是研究最透彻的细菌，因此其新陈代谢已被充分了解； (iv)对其基因组进行测序，以便获得 DNA 微阵列，并可以在基因组规模上进行代谢流分析。 除了研究成果外，参与这项研究的三位教授还将培训本科生和研究生，通过万维网提供该项目的具体成果，并将一些研究成果传播给不同的受众播客。