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

生命过程化学(CLP)项目的这一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(CHE-1058357)教授合作，对含铁和钴的氰基水合酶(NHASS)的机制进行时间分辨脉冲电子顺磁共振(EPR)光谱研究。氨酶催化氰化物立体定向水合反应的机理具有重要意义，该反应在制药工业中具有重要的实用价值和经济价值。同位素标记的(H-2，C-13，N-15)底物和NHase酶将通过快速冷冻-猝灭来孵育和收集，以捕获预稳态和稳态中间体，并将用脉冲EPR方法进行询问。模拟将被用来获得自旋哈密顿参数，从中可以阐明中间体的结构，并推导出机理。预计这项研究将为设计化学催化剂和工程酶铺平道路，这些催化剂和工程酶的性质可以针对特定的底物和期望的反应性质进行定制。这将大大增加药物和中间体的范围，这些药物和中间体依赖于能够有利可图地推向市场的立体选择性丁腈水合作用，并刺激对这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.