Procacci Barbara, Rutherford Samantha H, Greetham Gregory M, Towrie Michael, Parker Anthony W, Robinson Camilla V, Howle Christopher R, Hunt Neil T
Department of Chemistry and York Biomedical Research Institute, University of York, Heslington, York YO10 5DD, UK.
Department of Physics, University of Strathclyde, SUPA, 107 Rottenrow East, Glasgow G4 0NG, UK.
Spectrochim Acta A Mol Biomol Spectrosc. 2021 Mar 15;249:119319. doi: 10.1016/j.saa.2020.119319. Epub 2020 Dec 14.
Ultrafast 2D-IR spectroscopy is a powerful tool for understanding the spectroscopy and dynamics of biological molecules in the solution phase. A number of recent studies have begun to explore the utility of the information-rich 2D-IR spectra for analytical applications. Here, we report the application of ultrafast 2D-IR spectroscopy for the detection and classification of bacterial spores. 2D-IR spectra of Bacillus atrophaeus and Bacillus thuringiensis spores as dry films on CaF windows were obtained. The sporulated nature of the bacteria was confirmed using 2D-IR diagonal and off-diagonal peaks arising from the calcium dipicolinate CaDP·3HO biomarker for sporulation. Distinctive peaks, in the protein amide I region of the spectrum were used to differentiate the two types of spore. The identified marker modes demonstrate the potential for the use of 2D-IR methods as a direct means of spore classification. We discuss these new results in perspective with the current state of analytical 2D-IR measurements, showing that the potential exists to apply 2D-IR spectroscopy to detect the spores on surfaces and in suspensions as well as in dry films. The results demonstrate how applying 2D-IR screening methodologies to spores would enable the creation of a library of spectra for classification purposes.
超快二维红外光谱是理解溶液相中生物分子光谱和动力学的有力工具。最近的一些研究已开始探索信息丰富的二维红外光谱在分析应用中的效用。在此，我们报告超快二维红外光谱在细菌芽孢检测和分类中的应用。获得了萎缩芽孢杆菌和苏云金芽孢杆菌芽孢在CaF窗口上作为干膜的二维红外光谱。利用由吡啶二羧酸钙CaDP·3H₂O芽孢形成生物标志物产生的二维红外对角峰和非对角峰，证实了细菌的芽孢化性质。光谱中蛋白质酰胺I区域的独特峰用于区分两种芽孢。所识别的标记模式表明二维红外方法作为芽孢分类直接手段的潜力。我们结合分析二维红外测量的当前状况来探讨这些新结果，表明存在应用二维红外光谱检测表面、悬浮液以及干膜中的芽孢的潜力。结果表明，将二维红外筛选方法应用于芽孢将如何能够创建用于分类目的的光谱库。
