Department Applied Biology-Microbiology, University of Perugia, Borgo 20 Giugno, 74, I-06121 Perugia, Italy.
Analyst. 2011 Jun 7;136(11):2339-49. doi: 10.1039/c0an00515k. Epub 2011 Apr 15.
Fourier Transform InfraRed spectroscopy (FTIR) is an emerging technique in biology. One of the analytical problems inherent with this approach is the extreme variability typical of biological systems. Extrinsic fonts of variations are the growth media and the growth phase of the microbial culture. In order to gain knowledge on the relations between these two factors and the spectral variability, metabolomic changes in Saccharomyces cerevisiae cells during growth were analyzed with FTIR spectroscopy. Spectral variations reflected the changes occurring in all cellular compartments and components during the different growth stages and describe the metabolomic evolution of cell cultures during growth. Three different media were tested. Different mathematical and statistical treatments were used to describe and evaluate the changes occurring during growth. Variations were mainly concentrated in the exponential phase, whereas fewer changes occurred in stationary phase cells. The three different media caused different metabolomic behaviours during growth. The mathematical distance between the peak areas of amide I, amide II and of two distinct peaks in the carbohydrates region could give a synthetic description of growth and produced similar trends in all the conditions tested. The distance between the two carbohydrate peaks could be used to determine the mid exponential phase and the entrance in the stationary phase. Growth phase was shown to influence the sensitivity of a FTIR-based cell stress bioassay. The knowledge of the metabolomic variations during growth could be used in the future to optimize FTIR applications in microbiology.
傅里叶变换红外光谱(FTIR)是生物学中的一项新兴技术。这种方法固有的分析问题之一是生物系统的极端可变性。外在的变异来源是生长培养基和微生物培养的生长阶段。为了了解这两个因素与光谱可变性之间的关系,我们使用 FTIR 光谱法分析了酿酒酵母细胞在生长过程中的代谢组变化。光谱变化反映了在不同生长阶段所有细胞区室和成分中发生的变化,并描述了细胞培养在生长过程中的代谢组演变。我们测试了三种不同的培养基。使用不同的数学和统计处理方法来描述和评估生长过程中发生的变化。变化主要集中在指数生长期,而在静止期细胞中发生的变化较少。三种不同的培养基在生长过程中引起不同的代谢行为。酰胺 I、酰胺 II 和碳水化合物区域中两个不同峰的峰面积之间的数学距离可以对生长进行综合描述,并在所有测试条件下产生相似的趋势。两个碳水化合物峰之间的距离可用于确定指数中期和进入静止期。生长阶段表明会影响基于 FTIR 的细胞应激生物测定的灵敏度。未来,可以利用生长过程中的代谢组变化知识来优化 FTIR 在微生物学中的应用。