CAREER: Structural and mechanisctic studies of a novel group of oxygen-tolerant [FeFe] hydrogenases

职业：一组新型耐氧 [FeFe] 氢化酶的结构和机制研究

• 批准号: 1943748
• 负责人: Alexey Silakov
• 金额: $ 65万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1943748&HistoricalAwards=false
• 关键词: CAREER; Structural; mechanisctic; studies; novel

Hydrogen gas (H2) is growing in demand as an energy resource. It is currently produced from non-renewable sources, which is both costly and environmentally harmful. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Alexey Silakov from Pennsylvania State University at University Park to study a recently discovered bacterial enzyme that contains two iron atoms. This [FeFe] hydrogenase molecule speeds up chemical reactions so that they can efficiently produce H2 gas that can be used as an energy resource. Current, [FeFe] hydrogenases lose their activity upon exposure to even small amounts of oxygen (O2) from the air, hampering their industrial use for energy production and utilization. The [FeFe] hydrogenase employed by the Silakov group remains stable and active after O2 exposure. Dr. Silakov will apply structural and computational approaches to help explain why this [FeFe] hydrogenase is O2 tolerant, setting the stage for their use in environment-friendly and renewable hydrogen production and energy generation. This research project provides graduate and undergraduate students with opportunities to develop experimental and theoretical skills that are critical for their future careers in the sciences. In addition, Dr. Silakov develops lectures and hands-on training in research methods used to study enzymes as a part of the University’s biannual Bioinorganic Workshop. Finally, Dr. Silakov engages high-school students from under-served communities by developing curricular materials on the hydrogen-based economy and by providing laboratory experiences to foster their interest in pursuing careers in STEM fields. In this project, Dr. Silakov investigates a putative subclass of highly O2-tolerant [FeFe] hydrogenases. The discovery these [FeFe] hydrogenases sets a new paradigm for the stability of these highly efficient enzymes in the presence of oxygen. By studying structures of the O2-tolerant [FeFe] hydrogenases and physicochemical properties of the incorporated metallocofactors, this project establishes the connection between the enzymatic function, mechanisms of safe reaction with O2, and the structural determinants of specific phenotypes. The multidisciplinary research plan encapsulates advanced electron paramagnetic resonance and vibrational spectroscopies, electrochemistry, X-ray crystallography, molecular dynamic simulations, and quantum chemical calculations. This research formulates the structure-function relationships that are instrumental for engineering hydrogenases to perform a particular function and provides an in-depth understanding of how different components of the metal-containing enzymes work together. This project builds a rich platform to engage a diverse group of scientists from high-school to graduate students in STEM activities through a series of outreach activities.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.

氢气（H2）作为一种能源的需求正在增长。目前，它是由不可再生资源生产的，既昂贵又对环境有害。有了这个奖项，化学部的生命过程化学计划正在资助位于大学公园的宾夕法尼亚州立大学的Alexey Silakov博士研究最近发现的含有两个铁原子的细菌酶。 这种[FeFe]氢化酶分子加速了化学反应，因此它们可以有效地产生可用作能源的H2气体。 目前，[FeFe]氢化酶在暴露于来自空气的即使少量的氧气（O2）时也会失去活性，阻碍了它们在能源生产和利用方面的工业用途。Silakov组使用的[FeFe]氢化酶在O2暴露后保持稳定和活性。Silakov博士将应用结构和计算方法来帮助解释为什么这种[FeFe]氢化酶是O2耐受性的，为它们在环境友好和可再生氢气生产和能源生产中的应用奠定基础。该研究项目为研究生和本科生提供了发展实验和理论技能的机会，这些技能对他们未来的科学生涯至关重要。此外，Silakov博士还开发了用于研究酶的研究方法的讲座和实践培训，作为大学两年一度的生物无机研讨会的一部分。最后，Silakov博士通过开发关于氢基经济的课程材料和提供实验室经验来培养他们在STEM领域追求职业生涯的兴趣，来吸引来自服务不足社区的高中生。在这个项目中，Silakov博士研究了高度耐氧[FeFe]氢化酶的一个假定亚类。这些[FeFe]氢化酶的发现为这些高效酶在氧气存在下的稳定性提供了新的范例。通过研究耐氧[FeFe]氢化酶的结构和掺入的金属辅因子的理化性质，该项目建立了酶功能，与O2的安全反应机制和特定表型的结构决定因素之间的联系。多学科研究计划包括先进的电子顺磁共振和振动光谱，电化学，X射线晶体学，分子动力学模拟和量子化学计算。这项研究阐述了结构-功能关系，这些关系有助于工程氢化酶执行特定功能，并深入了解含金属酶的不同组分如何协同工作。该项目通过一系列的外展活动，搭建了一个丰富的平台，让从高中到研究生的各种科学家参与STEM活动。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Evidence of Atypical Structural Flexibility of the Active Site Surrounding of an [FeFe] Hydrogenase from Clostridium beijerinkii

• DOI: 10.1021/jacs.2c13458
• 发表时间: 2023-05-10
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Corrigan，Patrick S.;Majer，Sean H.;Silakov，Alexey
• 通讯作者: Silakov，Alexey

Using peptide substrate analogs to characterize a radical intermediate in NosN catalysis

• DOI: 10.1016/bs.mie.2022.02.008
• 发表时间: 2022-04-21
• 期刊: ADVANCES IN BIOMOLECULAR EPR
• 作者: Wang，Bo;Silakov，Alexey;Booker，Squire J.
• 通讯作者: Booker，Squire J.