Ash Philip A, Vincent Kylie A
Department of Chemistry, Inorganic Chemistry Laboratory, University of Oxford, South Parks Road, Oxford, OX1 3QR, UK.
Adv Biochem Eng Biotechnol. 2016;158:75-110. doi: 10.1007/10_2016_3.
A more complete understanding of bioelectrochemical interfaces is of increasing importance in both fundamental studies and biotechnological applications of proteins. Bioelectrochemical methods provide detailed information about the activity or rate of a process, but in situ spectroscopic methods are needed to gain direct structural insight into functionally relevant states. A number of methods have been reported that allow electrochemical and spectroscopic data to be collected from the same electrode, providing direct spectroscopic 'snapshots' of protein function, and here we focus on the application of infrared and Raman spectroscopies to the study of electrode-immobilised species. The ability to probe coordination at metal centres, protonation changes in amino acid side chains, reaction-induced changes in organic cofactors or substrates, protein orientation and subtle changes in protein secondary structure simultaneously, rapidly and at room temperature means that vibrational spectroscopic approaches are almost uniquely applicable to answering a wide range of questions in bioelectrochemistry.
在蛋白质的基础研究和生物技术应用中，更全面地理解生物电化学界面变得越来越重要。生物电化学方法可提供有关过程活性或速率的详细信息，但需要原位光谱方法才能直接深入了解功能相关状态的结构。已经报道了许多方法，这些方法能够从同一电极收集电化学和光谱数据，提供蛋白质功能的直接光谱“快照”，在这里我们重点关注红外光谱和拉曼光谱在固定于电极上的物种研究中的应用。能够在室温下同时、快速地探测金属中心的配位、氨基酸侧链的质子化变化、反应诱导的有机辅因子或底物的变化、蛋白质取向以及蛋白质二级结构的细微变化，这意味着振动光谱方法几乎是唯一适用于回答生物电化学中广泛问题的方法。
