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-0753702WOODNON HARMFUL细菌可用于为人类生产化学物质，其中包括燃料。 分子氢是一种环境清洁的燃料。因此，如果可以通过细菌有效地合成氢，那么我们可能会减少空气污染，减少对异物的依赖并避免温室气体（我们的过程形成的二氧化碳将被回收以种植质量并再次用于形成氢，以形成氢气，因此没有净发射量）。 这项研究将使氢以细菌的清洁形式使用简单糖的发酵生产。 在氢的发酵生产中，葡萄糖被转化为丙酮酸，然后转化为甲酸，然后转化为氢。 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，乳酸和乙醇的葡萄糖反应，将细菌细胞中的碳通量引导至氢形成。 为了实现这一目标，将使用一种系统生物学方法来生产更多的氢气，包括（i）通过去除不需要的途径来改变细菌的代谢，（ii）使用蛋白质工程来进化必要的酶，（iii）建模细菌细胞的所有代谢途径，从而可以产生更好的细菌。 我们选择了大肠杆菌作为模型系统，因为（i）所有3985个非致命的菌株大肠杆菌BW25113的非致命单突变； （ii）我们可以使用我们的新型，基于病毒的方法快速结合突变； （iii）大肠杆菌是研究最佳的细菌，因此它的新陈代谢得到了充分的理解。 （iv）它的基因组是测序的，因此可以使用DNA微阵列，并且可以在基因组规模进行代谢通量分析。 除研究结果外，参与这项研究的三位教授将培训本科生和研究生，通过全球网络，该项目的具体成果使访问权限，并将使用播客将一些研究结果传播给多样化的受众。