Enhanced Production of Advanced Biofuels through Model Guided Synthetic Biology

主要研究人员：Mark BlennerNumber：1437836该项目旨在通过了解糖是如何转化为生物柴油来提高基因工程菌生产生物柴油的能力。这将通过基于细胞的微生物监测来实现-S通过合成生物学的原理控制新陈代谢过程，该原理从整体上观察细胞内所有新陈代谢过程如何相互作用。该项目的第二个目标是控制这些代谢过程，以在不断变化的环境或工艺条件下保持较高的生物柴油产量。该项目的结果也可能适用于生产可持续燃料和化学品的其他代谢途径，使这些技术能够更确定、更少地从实验室转移到工业中。该项目的活动还将包括努力吸引南卡罗来纳州可能对科学、技术、工程和数学(STEM)职业感兴趣的妇女。教育和研究将结合起来，为已经在STEM学习的学生提供培训机会，并向可能对STEM更感兴趣的年轻女性进行当地外联。最后，这项研究将被整合到克莱姆森大学提供的一门关于蛋白质和代谢工程的新选修课中。本项目将使用简单的非稳态动力学模型来合理化蛋白质工程和基于合成生物学的对大肠杆菌生物柴油生产的改进。初步的模拟研究发现了由于途径不平衡造成的低效，并建议通过基因和蛋白质工程的努力可以实现更好的通量分布。将使用催化效率预计将提高生物柴油产量的酶。将使用定向进化来设计关键酶，以缓解途径瓶颈。蛋白质水平的反馈控制将被设计出来，并与遗传水平的反馈控制相结合，以允许途径通量保持高水平，尽管存在短期和长期的代谢扰动。基因和蛋白质水平的动态控制相结合，将使生物燃料和其他化学品生产系统能够经受住环境变化和扩大条件的干扰，而不会在产量和效率方面遭受重大损失。该项目的结果可能导致更通用的方法来平衡路径，而不依赖于显式的动力学或通量数据。该项目的活动还将包括努力吸引南卡罗来纳州可能对科学、技术、工程和数学(STEM)职业感兴趣的妇女。教育和研究将结合起来，为已经在STEM学习的学生提供培训机会，并向可能对STEM更感兴趣的年轻女性进行当地外联。最后，这项研究将被整合到克莱姆森大学提供的一门关于蛋白质和代谢工程的新选修课中。

Principal Investigator: Mark BlennerNumber: 1437836This project seeks to improve the production of biodiesel in a genetically engineered strain of bacteria by understanding how sugars are converted into biodiesel. This will be accomplished through cell-based monitoring of the microorganism?s metabolism control processes through the principles of synthetic biology, which look holistically at how all metabolic processes within a cell interact with one another. A second aim of this project is control these metabolic processes to maintain high biodiesel production rates under changing environmental or process conditions. The results of this project may also apply to other metabolic pathways that produce sustainable fuels and chemicals, allowing transfer of these techniques from the laboratory to industry with more certainty and fewer complications. The project activities will also include efforts to engage women potentially interested in science, technology, engineering, and mathematics (STEM) careers in the state of South Carolina. Education and research will be integrated by providing training opportunities for students already in STEM, and local outreach to young women who might become more interested in STEM. Finally, this research will be integrated into a new elective course on protein and metabolic engineering offered at Clemson University. Technical DescriptionThis project will use simple unsteady-state kinetic models to rationalize protein engineering and synthetic biology based improvements to E. coli biodiesel production. Preliminary modeling studies identify inefficiencies due to pathway imbalance, and suggest that better flux distributions can be achieved through genetic and protein engineering efforts. Enzymes with catalytic efficiencies predicted to improve biodiesel production will be used. Critical enzymes will be engineered using directed evolution to relieve pathway bottlenecks. Protein-level feedback control will be engineered and combined with genetic level feedback control to allow pathway fluxes to remain high in spite of short and long time-scale metabolic perturbations. The combination of genetic and protein-level dynamic control will allow biofuel and other chemical producing systems to withstand perturbations from environmental variation and scale-up conditions without suffering large losses in yield and efficiency. The results of this project may lead to more general methods for balancing pathways that does not rely on explicit kinetic or flux data. The project activities will also include efforts to engage women potentially interested in science, technology, engineering, and mathematics (STEM) careers in the state of South Carolina. Education and research will be integrated by providing training opportunities for students already in STEM, and local outreach to young women who might become more interested in STEM. Finally, this research will be integrated into a new elective course on protein and metabolic engineering offered at Clemson University.