CAREER: Chemically-Enabled Strategies for the Discovery and Characterization of Novel Enzymatic Function

职业：新酶功能的发现和表征的化学策略

• 批准号: 1454007
• 负责人: Emily Balskus
• 金额: $ 65万
• 依托单位: Harvard University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-15 至 2020-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1454007&HistoricalAwards=false
• 关键词: CAREER; Chemically; Enabled; Strategies; Discovery

Microorganisms are not only the most abundant form of life on Earth, but they also possess the greatest diversity of metabolic functions. Gaining access to their collective chemical arsenal would significantly enhance our ability to produce essential small molecules ranging from commodity chemicals to life-saving pharmaceuticals. Recent advances in DNA sequencing technologies have delivered a wealth of microbial genomes that provide an unprecedented opportunity for the discovery of new enzymes. The goal of this project is to identify unusual and potentially useful enzymatic chemistry encoded in these genomic data. The discovery of previously uncharacterized enzymatic activity and its introduction into our reaction repertoire will ultimately lead to more efficient and environmentally friendly routes for small molecule synthesis. The specific C-C bond forming enzymes described in this work will be important additions to the synthetic toolkits of both organic chemists and synthetic biologists. These research efforts will also be integrated into a new laboratory-based freshman seminar course that will engage students in the discovery of new enzymes from human gut microbes. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Emily Balskus from Harvard University to discover and characterize enzymes that mediate an unusual C-C bond-forming macrocyclizing dimerization step used in the biosynthesis of the cylindrocyclophane family of cyanobacterial natural products. Using a genome mining approach that is guided by chemical knowledge, the PI has uncovered the biosynthetic gene cluster encoding the enzymes responsible for this reaction. The functions and mechanisms of these enzymes will now be elucidated using in vivo and in vitro biochemical characterization methods, an effort that will be enabled by chemical synthesis of crucial substrates. Important mechanistic insights will also be obtained by solving the crystal structures of the enzymes in collaboration with structural biologists. The central educational objective of this proposal is to create a new research-based course for first-year undergraduates focused on using a screening approach to discover novel enzymes from the human gut microbiota. This class will bring the most exciting aspects of scientific training to the forefront of the college curriculum, encouraging interest in further undergraduate research and scientific careers. Overall, the work detailed in this proposal represents an integrated research and education effort at the interface of chemistry and biology.

微生物不仅是地球上最丰富的生命形式，而且还具有代谢功能最大的多样性。获得其集体化学库的获取将显着增强我们生产从商品化学物质到挽救生命的药物等必不可少的小分子的能力。 DNA测序技术的最新进展提供了大量的微生物基因组，为发现新酶提供了前所未有的机会。该项目的目的是确定这些基因组数据中编码的异常和潜在有用的酶学化学。以前未表征的酶活性的发现及其引入我们的反应曲目最终将导致小分子合成的更有效且对环境友好的途径。这项工作中描述的特定的C-C键形成酶将是有机化学家和合成生物学家的合成工具包的重要补充。这些研究工作还将集成到新的基于实验室的新生研讨会课程中，该课程将吸引学生从人类肠道微生物中发现新酶。通过该奖项，化学过程的化学过程计划正在为哈佛大学的艾米莉·巴尔斯库斯（Emily Balskus）博士提供资金，以发现和表征酶，这些酶介导了在生物合成中使用cylindrocyclopophane族的生物合成中使用的异常C-C键形成的宏环化二聚步骤。使用化学知识指导的基因组挖掘方法，PI发现了编码负责该反应的酶的生物合成基因簇。现在，将使用体内和体外生化特征方法阐明这些酶的功能和机制，这将通过化学合成至关重要的底物来实现这一努力。还将通过与结构生物学家合作解决酶的晶体结构来获得重要的机械见解。该提案的核心教育目标是为一年级的大学生创建一个新的基于研究的课程，专注于使用筛查方法来发现人类肠道菌群的新酶。该课程将使科学培训的最激动人心的方面成为大学课程的最前沿，鼓励人们对进一步的本科研究和科学职业的兴趣。总体而言，本提案中详细介绍的工作代表了化学和生物学界面的一体综合研究和教育工作。