Unravelling enzymatic hydrogen production mechanisms with ultrafast 2D-IR spectroscopy

利用超快二维红外光谱揭示酶促产氢机制

• 批准号: 2107430
• 金额: --
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2107430
• 关键词: Unravelling; enzymatic; hydrogen; production; mechanisms

The hydrogenases are families of metalloenzymes that efficiently catalyse the reversible cleavage of H2 into protons and electrons and so offer ideal prototype catalysts for sustainable energy generation involving H2. Before we can develop technological materials based upon the hydrogenases however, a detailed understanding of the mechanism of H2 production and usage is required. This multidisciplinary project will combine ultrafast 2D-IR spectroscopy (Hunt) with electrochemical and biochemical methods (Parkin) to understand structural changes and the role of dynamic processes in the action of these important enzymes. 2D-IR is a new ultrafast laser spectroscopy method that spreads the IR spectrum of a molecule over a second frequency axis [1]. This multidimensionality allows access to a wealth of new spectral information but the most powerful aspect of 2D-IR is the ability to determine molecular structure and changes in that structure with a time resolution of 100 fs (10-13 s). This opens up the possibility of observing reaction steps, solvent motion or H-bond vibrations that are central to the hydrogenase mechanism in real time. This is highly complementary to the insight into ultrafast electron movement in hydrogenases which can has been obtained in the Parkin group using Fourier transform large amplitude alternating current voltammetry [2]. Although synthetic models of the hydrogenases have been studied with 2D-IR [3,4], this collaboration offers the unique opportunity to compare these results with the full enzyme systems for the first time. By studying a range of site mutations and using electrochemical methods to prepare catalytic intermediates we will use 2D-IR to understand the way in which the protein scaffold interacts with the active site to influence or control the enzyme mechanism. This is a particularly important issue since biomimetic model compounds lacking the protein scaffold show very low catalytic efficiency, emphasising its importance [4], but the precise molecular function that it plays in the enzyme cycle is unknown.(1) Hunt, N. T. Chem Soc Rev 2009, 38, 1837.(2) Adamson, H.; Robinson, M.; Wright, J. J.; Flanagan, L. A.; Walton, J.; Elton, D.; Gavaghan, D. J.; Bond, A. M.; Roessler, M. M.; Parkin, A. Journal of the American Chemical Society 2017, 139, 10677.(3) Fritzsch, R.; Brady, O.; Adair, E.; Wright, J. A.; Pickett, C. J.; Hunt, N. T. Journal of Physical Chemistry Letters 2016, 7, 2838.(4) Frederix, P. W. J. M.; Adamczyk, K.; Wright, J. A.; Tuttle, T.; Ulijn, R. V.; Pickett, C. J.; Hunt, N. T. Organometallics 2014, 33, 5888.

氢化酶是金属酶的家族，其有效地催化H2可逆裂解成质子和电子，因此为涉及H2的可持续能源产生提供理想的原型催化剂。然而，在我们开发基于氢化酶的技术材料之前，需要详细了解H2生产和使用的机制。这个多学科项目将联合收割机超快2D-IR光谱（亨特）与电化学和生物化学方法（帕金）相结合，以了解结构变化和动态过程在这些重要酶的作用。2D-IR是一种新的超快激光光谱学方法，其将分子的IR光谱扩展到第二频率轴上[1]。这种多维性允许访问丰富的新光谱信息，但2D-IR最强大的方面是能够以100 fs（10-13 s）的时间分辨率确定分子结构和该结构的变化。这开辟了观察反应步骤，溶剂运动或氢键振动的可能性，在真实的时间是氢化酶机制的核心。这是对氢化酶中超快电子运动的深入了解的高度补充，这可以在Parkin组中使用傅立叶变换大幅度交流伏安法[2]获得。虽然氢化酶的合成模型已经用2D-IR [3，4]进行了研究，但这项合作首次提供了将这些结果与完整酶系统进行比较的独特机会。通过研究一系列位点突变和使用电化学方法制备催化中间体，我们将使用2D-IR来了解蛋白质支架与活性位点相互作用以影响或控制酶机制的方式。这是一个特别重要的问题，因为缺乏蛋白质支架的仿生模型化合物显示出非常低的催化效率，强调了其重要性[4]，但它在酶循环中发挥的精确分子功能是未知的。(1)Hunt，N. T. Chem Soc Rev 2009，38，1837。(2)Adamson，H.;罗宾逊; Wright，J. J.;弗拉纳根湖一、Walton，J.; Elton，D.; Gavaghan，D. J.道：邦德，A. M.; Roessler，M. M.;帕金，A。Journal of the American Chemical Society 2017，139，10677. (3)Fritzsch，R.;布雷迪，O.; Adair，E.; Wright，J. A.;皮克特角J.道：Hunt，N. T. Journal of Physical Chemistry Letters 2016，7，2838. (4)Frederix，P. W. J. M.; Adamczyk，K.; Wright，J. A.; Tuttle，T.;乌利因河五、皮克特角J.道：Hunt，N. T.有机金属2014，33，5888。