CAREER: Engineering redox metabolism using unnatural cofactors

可以使用微生物从可再生资源中合成化学品。这种方法可以帮助化学工业变得更可持续。广泛实施这一战略的一个主要障碍是电池的生产效率较低。克服这一问题的标准方法是改变细胞新陈代谢中的碳流。在这个项目中，通过电池的电子流量将被调整。这将通过创造人造电子穿梭分子来实现，这种分子被称为辅因子。这是一个创新的战略。它可以极大地提高利用微生物制造化学品的效率。外联活动将与研究任务结合起来。因此，该项目代表着一种尝试：(I)允许生物制造更好地满足国家对能源、食品、商品和药品的需求；(Ii)促进STEM的本科生和研究生教育；以及(Iii)激励奥兰治县的女高中生从事工程工作。分解代谢和合成代谢共存而不受干扰，因为每个分解代谢都有一个单独的氧化还原辅因。第三，独立的氧化还原辅因子可以使特定的途径与核心代谢隔绝。通过建立新的氧化还原辅因子，并在产物途径中进化酶以唯一需要该辅因子，可以提高生产率。该项目将确定烟酰胺单核苷酸(NMN)为第三个氧化还原辅因子。这将通过一个分三步走的实验计划来实现。首先，将开发用于NMN相关电子电路工程的高通量选择平台。然后，将构建能够高产NMN的大肠杆菌和酿酒酵母菌株。最后，依赖于NMN的甲酸脱氢酶将被设计成提供独立于宿主代谢背景的NMN还原能力。成功完成这些目标将产生两种与工业相关的菌株，能够作为依赖NMN的途径和产品的底盘。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Chemicals can be synthesized from renewable resources using microbes. This approach could help the chemical industry become more sustainable. A major hurdle to the widespread implementation of this strategy is low production efficiency on the part of the cells. The standard approach to overcome this has been to modify the flow of carbon through cell metabolism. In this project, the flow of electrons through the cell will be adjusted. This will be accomplished by creating artificial electron shuttle molecules, known as cofactors. This is an innovative strategy. It could greatly increase the efficiency of using microbes to manufacture chemicals. Outreach activities will be integrated with the research tasks. As a result, the project represents an attempt to (i) allow biomanufacturing to better meet the Nation's needs for energy, food, commodities, and medicine; (ii) contribute to undergraduate and graduate education in STEM; and (iii) motivate female high school students in Orange County to pursue a career in engineering.Catabolism and anabolism co-exist without interference because each has a separate redox cofactor. A third, independent redox cofactor may enable a specific pathway to be insulated from core metabolism. Increased productivity might be achieved by establishing the new redox cofactor and evolving enzymes in the product pathway to require that cofactor exclusively. This project will establish nicotinamide mononucleotide (NMN) as the third redox cofactor. This will be accomplished through a three-step experimental plan. First, high-throughput selection platforms for engineering of NMN-dependent electron circuits will be developed. Then, strains of E. coli and S. cerevisiae capable of overproduction of NMN will be constructed. Finally, NMN-dependent formate dehydrogenase will be engineered to deliver NMN reducing power independently of the host's metabolic background. Successful completion of these objectives will result in two industrially-relevant strains capable of acting as a chassis for NMN-dependent pathways and products.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.