Collaborative Research: Mechanistic Studies on Fe-type Nitrile Hydration Catalysts

丁腈广泛用于生产特殊化学品，如聚酰胺尼龙。腈水合酶（nase）在中性水中的环境温度下催化腈水合成酰胺。目前用于将腈水化成酰胺的工业条件使用强酸或强碱，并且通常与要生产的化合物的敏感结构不相容。因此，nase作为生物催化剂在制备有机化学中引起了极大的兴趣。尽管nase酶在工业上具有重要意义，但人们对其工作原理的细节仍然知之甚少。本研究计划的目的是填补这一知识空白。这项工作的跨学科性质将作为一个平台，增加女性和传统上代表性不足的研究生和本科生参与生物物理学、化学和生物化学的科学事业。作为拟议项目的一部分，两位PI将参与SMART项目，该项目指导密尔沃基地区公立高中学生进行研究项目。有了这个奖项，化学部门的生命过程化学项目正在资助博士。来自马凯特大学的Richard C. Holz和Brian Bennett来阐明nase酶的催化机制。将采用跨学科的方法，包括动力学，光谱学，生物化学，计算化学，以及x射线和中子晶体学。提出的实验方法旨在解决三个关键问题：i)什么是腈水化的过渡态结构？ii)在整个反应过程中，催化重要物质的质子化状态是什么？以及iii)在所谓的真核nase中是什么控制着金属中心的组装？这项研究的成功完成将极大地促进腈基手性药物和工业上重要的特种化学品的智能设计和制造，具有比目前更高的合成灵活性和更低的环境影响。

Nitriles are used extensively in the production of specialty chemicals like the polyamide nylon. Nitrile hydratases (NHases) catalyze the hydration of nitriles to amides at ambient temperatures in neutral water. The currently employed industrial conditions for the hydration of nitriles to amides use either strong acids or bases, and are often incompatible with the sensitive structures of the compounds to be produced. NHases have therefore attracted substantial interest as biocatalysts in preparative organic chemistry. Despite the industrial importance of NHase enzymes, the details of how they work remain poorly understood. The objective of this research proposal is to fill this knowledge gap. The interdisciplinary nature of the effort will serve as a platform to increase the participation of women and classically underrepresented graduate and undergraduate students in scientific careers in biophysics, chemistry, and biochemistry. As part of the proposed project, both PI's will participate in the SMART program that mentors Milwaukee area public high school students in research projects.With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Drs. Richard C. Holz and Brian Bennett from Marquette University to elucidate the catalytic mechanism of NHase enzymes. An interdisciplinary approach will be used that will incorporate kinetics, spectroscopy, biochemistry, computational chemistry, and both X-ray and neutron crystallography. The proposed experimental approach is designed to address three key questions: i) What is the transition-state structure of nitrile hydration? ii) What are the protonation states of catalytically important species throughout the reaction? and iii) What controls metallocenter assembly in a purported eukaryotic NHase? The successful completion of this research will greatly benefit society by facilitating the intelligent design and manufacture of nitrile-based chiral pharmaceuticals and industrially important specialty chemicals, with enhanced synthetic flexibility and lower environmental impact, than at present.