Collaborative Research: CAS: Mechanistic Studies on Fe-Type Nitrile Hydration Catalysts

合作研究：CAS：铁型腈水合催化剂的机理研究

• 批准号: 2204023
• 负责人: Brian Bennett
• 金额: $ 39.11万
• 依托单位: Marquette University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2204023&HistoricalAwards=false
• 关键词: Collaborative; Research; CAS; Mechanistic; Studies

With the support of the Chemistry of Life Processes program in the Division of Chemistry, Professors Brian Bennett and Adam Fiedler from Marquette University and Professor Richard Holz from the Colorado School of Mines will investigate the chemical function of the enzyme nitrile hydratase (NHase), which catalyzes the reaction of nitrile compounds with water (hydration) to give the corresponding amides. NHases hydrate a range of natural and synthetic nitriles with strict selectivity under mild conditions. As such, these enzymes have several industrial and pharmaceutical applications, including the removal of chemicals and pesticides from the environment. To expand the biocatalytic potential of NHases, a better understanding of the catalytic mechanism is essential. Professors Bennett, Fiedler and Holz will employ a combination of experimental and computational approaches to detect and probe intermediates generated during the hydration reaction. In addition, the assembly of the enzyme active site, which contains a cobalt or iron ion, will be examined. The project will provide exceptional training for undergraduate and graduate-level scientists. The emphasis on green chemistry and biophysical methods will serve as the basis for community engagement activities. The award will also enhance the research and educational infrastructures at both Marquette University and the Colorado School of Mines. This research project seeks to gain molecular-level insights into NHase catalysis through a combination of kinetic, spectroscopic, biochemical, computational, and X-ray crystallographic methods. The studies will (i) test competing prevailing mechanistic hypotheses and refine the catalytic mechanism by interrogating observed intermediates, (ii) determine the catalytic function of proton transfer and the protonation states of ionizable groups in the active site, and (iii) elucidate steps in the maturation process of the metal-based (Fe or Co) active sites. The insights gained from the studies are expected to aid in the development of green catalytic approaches to hydrolytic reactions in nature and industry.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.

在化学系生命过程化学项目的支持下，马凯特大学的 Brian Bennett 和 Adam Fiedler 教授以及科罗拉多矿业学院的 Richard Holz 教授将研究腈水合酶 (NHase) 的化学功能。催化腈化合物与水的反应（水合），生成相应的酰胺。 NHases 在温和条件下以严格的选择性水合一系列天然和合成的腈。 因此，这些酶具有多种工业和制药应用，包括去除环境中的化学品和农药。 为了扩大 NHases 的生物催化潜力，更好地了解催化机制至关重要。 Bennett、Fiedler 和 Holz 教授将采用实验和计算相结合的方法来检测和探测水合反应过程中产生的中间体。 此外，还将检查含有钴或铁离子的酶活性位点的组装。 该项目将为本科生和研究生水平的科学家提供特殊的培训。 对绿色化学和生物物理方法的重视将作为社区参与活动的基础。 该奖项还将加强马凯特大学和科罗拉多矿业学院的研究和教育基础设施。该研究项目旨在通过结合动力学、光谱、生物化学、计算和 X 射线晶体学方法，获得对 NHase 催化的分子水平见解。 这些研究将（i）测试相互竞争的普遍机制假设并通过询问观察到的中间体来完善催化机制，（ii）确定质子转移的催化功能和活性位点中可电离基团的质子化状态，以及（iii）阐明金属基（Fe 或 Co）活性位点的成熟过程。从研究中获得的见解预计将有助于开发自然和工业中水解反应的绿色催化方法。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力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

Examination of the Catalytic Role of the Axial Cystine Ligand in the Co-Type Nitrile Hydratase from Pseudonocardia thermophila JCM 3095

轴向胱氨酸配体在嗜热假诺卡氏菌 JCM 3095 共型腈水合酶中的催化作用的检查

• DOI: 10.3390/catal11111381
• 发表时间: 2021-11
• 期刊: Catalysts
• 影响因子: 3.9
• 作者: Ogutu, Irene R.;St. Maurice, Martin;Bennett, Brian;Holz, Richard C.
• 通讯作者: Holz, Richard C.

Identification of an Intermediate Species along the Nitrile Hydratase Reaction Pathway by EPR Spectroscopy

通过 EPR 光谱鉴定腈水合酶反应途径的中间物质

• DOI: 10.1021/acs.biochem.1c00574
• 发表时间: 2021-12
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Karunagala Pathiranage, Wasantha Lankathilaka;Gumataotao, Natalie;Fiedler, Adam T.;Holz, Richard C.;Bennett, Brian
• 通讯作者: Bennett, Brian