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

在化学系生命过程化学项目的支持下，马奎特大学的Brian Bennett教授和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，该酶催化腈化合物与水(水合)的反应，得到相应的酰胺类化合物。氨酶在温和的条件下水合一系列具有严格选择性的天然和合成腈。因此，这些酶有几个工业和制药应用，包括从环境中去除化学物质和杀虫剂。为了扩大NHase的生物催化潜力，更好地了解其催化机理是必不可少的。贝内特、菲德勒和霍尔茨教授将采用实验和计算相结合的方法来检测和探测在水合反应中产生的中间产物。此外，还将检查含有钴或铁离子的酶活性部位的组装。该项目将为本科生和研究生级别的科学家提供特殊的培训。强调绿色化学和生物物理方法将成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。这项研究项目试图通过结合动力学、光谱、生化、计算和X射线结晶学方法从分子水平上深入了解NHase的催化作用。这些研究将(I)检验相互竞争的主流机理假说，并通过询问观察到的中间体来完善催化机理，(Ii)确定质子转移的催化作用和活性中心中可电离基团的质子化状态，以及(Iii)阐明金属基(Fe或Co)活性中心成熟过程中的步骤。从研究中获得的见解有望帮助开发用于自然界和工业中的水解反应的绿色催化方法。这一奖项反映了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。

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.