CAREER: Connecting eukaryotic electron transfer components to nitrogenase using a bacterial chassis

职业：使用细菌底盘将真核电子传递组件连接到固氮酶

• 批准号: 2338085
• 负责人: Kathryn Fixen
• 金额: $ 103.62万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2029-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2338085&HistoricalAwards=false
• 关键词: CAREER; Connecting; eukaryotic; electron; transfer

Important advancements in our understanding of biological nitrogen fixation, bolstered by emerging synthetic biology tools, suggest we are closer than ever to engineering plants to fix nitrogen. Engineering plants to fix nitrogen could improve the sustainability of the bioeconomy. However, we lack the knowledge to predict the behavior and specificity of electron carriers such as ferredoxin, which are needed to power nitrogenase. There is a critical need to test eukaryotic electron transfer components for their ability to interact with nitrogenase and measure how changes in the cellular redox environment sustain electron flow. The overall objective of the research proposed here is to use a bacterial chassis to rapidly define how eukaryotic electron transfer components can participate in electron delivery to nitrogenase and invent a powerful platform for evolution of synthetic electron flow pathways. The research aims synergize with educational goals by incorporating a semester-long project that focuses on bioengineering nitrogen fixation into existing courses using a novel culturally responsive pedagogical framework.The central hypothesis for the project is that electron transfer to nitrogenase is one of the primary constraints preventing introduction of this enzyme into eukaryotic systems, but it is possible to select for variants in eukaryotic electron transfer components to overcome this bottleneck. To test this hypothesis, the investigator proposes to develop a new tool to analyze electron flow to nitrogenase. This tool will use a bacterial chassis to test eukaryotic electron transfer components at physiological levels and evolve these electron transfer components for enhanced electron flow to nitrogenase. Such a contribution would be significant because it would enable more accurate predictions regarding nitrogenase functionality within plant organelles and would establish a robust platform for optimizing electron transfer to nitrogenase. This would not only make the goal of engineering plants to fix nitrogen more attainable, but it would also further our understanding of the determinants of electron flow and how electron transfer pathways can be optimized for biotechnological purposes. This project is supported by the Systems and Synthetic Biology Cluster of the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

通过新兴的合成生物学工具支持生物氮固定的重要进步，这表明我们比以往任何时候都更接近工厂以固定氮。固定氮的工程工厂可以改善生物经济的可持续性。但是，我们缺乏预测电子载体（例如铁氧还蛋白）的行为和特异性的知识，这些电子载体是为氮酶供电的。迫切需要测试真核电子转移成分与氮相互作用并测量细胞氧化还原环境中的变化如何维持电子流的能力。这里提出的研究的总体目的是使用细菌底盘快速定义真核电子传递成分如何参与电子传递到氮酶，并为合成电子流途径进化的强大平台发明了强大的平台。 The research aims synergize with educational goals by incorporating a semester-long project that focuses on bioengineering nitrogen fixation into existing courses using a novel culturally responsive pedagogical framework.The central hypothesis for the project is that electron transfer to nitrogenase is one of the primary constraints preventing introduction of this enzyme into eukaryotic systems, but it is possible to select for variants in eukaryotic electron transfer要克服这种瓶颈的组件。为了检验这一假设，研究者建议开发一种新工具来分析电子流向氮酶。该工具将使用细菌底盘在生理水平上测试真核电子转移成分，并进化这些电子转移成分，以增强电子流向氮酶。这样的贡献将是重要的，因为它可以对植物细胞器中的氮酶功能进行更准确的预测，并建立一个可靠的平台，以优化电子转移到氮酶。这不仅将使工厂更具固定氮的目的，而且还将进一步了解我们对电子流的决定因素以及如何为生物技术目的优化电子传输途径。该项目得到了分子和蜂窝生物科学划分的系统和合成生物学集群的支持。该奖项反映了NSF的法定任务，并使用基金会的知识分子优点和更广泛的影响审查标准，认为值得通过评估来获得支持。