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

这项生命过程化学（CLP）项目的合作研究奖支持威斯康星医学院生物物理系的Brian Bennett教授与芝加哥洛约拉大学的Richard C. Holz教授（CHE-1058357）合作，对含铁和含钴腈水合酶（nases）的机制进行时间分辨脉冲电子顺磁共振（EPR）光谱研究。nase对腈的立体特异水化反应机理的研究具有重要意义，在制药工业中具有重要的实用和经济意义。同位素标记的（H-2, C-13, N-15）底物和nase酶将通过快速冻灭培养和收集，以捕获预稳态和稳态中间体，这些将通过脉冲EPR方法进行检测。模拟将用于获得自旋哈密顿参数，从中可以阐明中间体的结构，并推断出一种机制。预计这项研究将为化学催化剂和工程酶的设计铺平道路，这些酶的性质可以根据特定的底物和所需的反应性质进行定制。这将大大增加依赖于立体选择性腈水化的药物和中间体的范围，这些药物和中间体可以有利可图地推向市场，并刺激对该类新化合物的进一步研究。

This collaborative research award in the Chemistry of Life Processes (CLP) program supports work by Prof. Brian Bennett of the Department of Biophysics at the Medical College of Wisconsin to carry out a time-resolved pulsed-electron paramagnetic resonance (EPR) spectroscopic study of the mechanism of iron- and cobalt-containing nitrile hydratases (NHases), in collaboration with Prof. Richard C. Holz (CHE-1058357) at Loyola University, Chicago. The mechanism of the stereospecific hydration of nitriles by NHases is of fundamental interest, and the reaction is of great practical and economic importance in the pharmaceutical industry. Isotopically labeled (H-2, C-13, N-15) substrates and NHase enzyme will be incubated and collected by rapid-freeze-quench to trap pre-steady-state and steady-state intermediates, and these will be interrogated by pulsed EPR methods. Simulations will be used to obtain spin-Hamiltonian parameters from which the structures of the intermediates can be elucidated, and a mechanism deduced.It is anticipated that this research will pave the way for the design of chemical catalysts and engineered enzymes whose properties can be tailored toward specific substrates and desired reaction properties. This will greatly increase the range of pharmaceuticals and intermediates that rely on stereoselective nitrile hydration that can be profitably brought to market, and stimulate further research into new compounds of this class.