CAREER: Engineering redox metabolism using unnatural cofactors

职业：使用非天然辅助因子工程氧化还原代谢

• 批准号: 1847705
• 负责人: Han Li
• 金额: $ 50.22万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1847705&HistoricalAwards=false
• 关键词: CAREER; Engineering; redox; metabolism; using

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.

可以使用微生物从可再生资源合成化学品。这种方法可以帮助化学工业变得更加可持续。该策略广泛实施的主要障碍是细胞的生产效率较低。克服这一点的标准方法是通过细胞代谢来改变碳的流动。在这个项目中，将调整电子通过电池的流动。这将通过创建人造电子班车分子（称为辅助因子）来实现。这是一种创新的策略。它可以大大提高使用微生物制造化学物质的效率。外展活动将与研究任务集成。结果，该项目代表了（i）允许生物制造能够更好地满足国家对能源，食品，商品和医学的需求； （ii）为STEM的本科和研究生教育做出了贡献； （iii）激励奥兰治县的女高中生从事工程职业。稳定症和代谢性共存而不会干扰，因为每个人都有一个单独的氧化还原辅助因子。第三个独立的氧化还原辅助因子可以使特定的途径与核心代谢隔离。通过在产品途径中建立新的氧化还原辅因子和不断发展的酶来实现生产率提高，以仅要求辅助因子。该项目将建立烟酰胺单核苷酸（NMN）作为第三个氧化还原辅助因子。这将通过三步的实验计划来完成。首先，将开发用于NMN依赖电子电路工程的高通量选择平台。然后，将构建能够过度生产NMN的大肠杆菌和酿酒酵母的菌株。最后，将设计依赖NMN的甲酸脱氢酶，以独立于宿主的代谢背景来提供NMN降低功率。这些目标的成功完成将导致两种与工业相关的菌株，能够充当NMN依赖性途径和产品的底盘。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛的审查标准通过评估来进行评估的。

项目成果

Metabolic engineering of Escherichia coli for optimized biosynthesis of nicotinamide mononucleotide, a noncanonical redox cofactor

• DOI: 10.1186/s12934-020-01415-z
• 发表时间: 2020-07-27
• 期刊: MICROBIAL CELL FACTORIES
• 影响因子: 6.4
• 作者: Black, William B.;Aspacio, Derek;Li, Han
• 通讯作者: Li, Han

Directed evolution of phosphite dehydrogenase to cycle noncanonical redox cofactors via universal growth selection platform.

• DOI: 10.1038/s41467-022-32727-w
• 发表时间: 2022-08-26
• 期刊: Nature communications
• 影响因子: 16.6

In Vivo, High-Throughput Selection of Thermostable Cyclohexanone Monooxygenase (CHMO)

• DOI: 10.3390/catal10080935
• 发表时间: 2020-08
• 期刊: Catalysts (Basel, Switzerland)
• 作者: Sarah Maxel;Linyue Zhang;Edward King;Ana Paula Acosta;R. Luo;Han Li