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Raman Deuterium Isotope Probing Reveals Microbial Metabolism at the Single-Cell Level
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-11-28 00:00:00 , DOI: 10.1021/acs.analchem.7b03461 Jiabao Xu 1 , Di Zhu 2 , Aliyu D. Ibrahim 3 , Christopher C. R. Allen 3 , Christopher M. Gibson 4 , Patrick W. Fowler 4 , Yizhi Song 1 , Wei E. Huang 1, 2
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-11-28 00:00:00 , DOI: 10.1021/acs.analchem.7b03461 Jiabao Xu 1 , Di Zhu 2 , Aliyu D. Ibrahim 3 , Christopher C. R. Allen 3 , Christopher M. Gibson 4 , Patrick W. Fowler 4 , Yizhi Song 1 , Wei E. Huang 1, 2
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We illustrate that single-cell Raman microspectroscopy, coupled with deuterium isotope probing (Raman-DIP), provides a culture-independent and nondestructive approach to probe metabolic pathways of carbon substrates at the single-cell level. We found a distinguishable C–D vibration band at 2070–2300 cm–1 in single-cell Raman spectra (SCRS) when Escherichia coli used deuterated glucose and Pseudomonas sp. used deuterated naphthalene as sole carbon sources. The intensity of the C–D band is proportional to the extent of deuteration in the carbon source, and as little as 5% deuteration can be distinguished by analysis of SCRS. It suggests that Raman-DIP could be used to semiquantitatively and sensitively indicate the metabolism of deuterated carbon source in microbes. A lower lipid conversion rate of deuterated naphthalene compared to that of deuterated glucose was observed, presumably owing to different anabolic pathways and membrane alteration. Apart from the C–D band shift from C–H, SCRS also reveal several isotopic shifts of the phenylalanine band, of which the positions correlate well with a computational model. A reduction in phenylalanine deuteration in Pseudomonas sp. compared to that in E. coli is due to the dilution effect of different pathways of phenylalanine biosynthesis in Pseudomonas sp. Collectively, we demonstrate that Raman-DIP can not only indicate metabolic activity using deuterated carbon sources but also reveal different metabolic pathways by analyzing SCRS. By harnessing such low-cost and versatile deuterated substrates, Raman-DIP has the potential to probe a wide range of metabolic pathways and functions at the single-cell level.
中文翻译：
拉曼氘同位素探测揭示了单细胞水平的微生物代谢
我们举例说明，单细胞拉曼光谱，结合氘同位素探测（Raman-DIP），提供了一种独立于培养且非破坏性的方法来探测单细胞水平上碳底物的代谢途径。当大肠杆菌使用氘代葡萄糖和假单胞菌时，我们在单细胞拉曼光谱（SCRS）中发现了2070–2300 cm –1处明显的C–D振动带。sp。使用氘代萘作为唯一碳源。C–D谱带的强度与碳源中氘的程度成正比，而通过SCRS分析可以分辨出只有5％的氘。这表明拉曼-DIP可用于半定量和灵敏地指示氘代碳源在微生物中的代谢。与氘化葡萄糖相比，氘化萘的脂质转化率较低，这可能是由于合成代谢途径和膜改变不同所致。除了从C–H到C–D的频移外，SCRS还揭示了苯丙氨酸带的几种同位素频移，其位置与计算模型很好地相关。减少了假单胞菌sp的苯丙氨酸氘化。相比于大肠杆菌归因于假单胞菌属物种中苯丙氨酸生物合成不同途径的稀释作用。集体地，我们证明拉曼-DIP不仅可以表明使用氘代碳源的代谢活性，而且还可以通过分析SCRS揭示不同的代谢途径。通过利用这种低成本，通用的氘代底物，拉曼DIP有望在单细胞水平上探索广泛的代谢途径和功能。
更新日期：2017-11-28
中文翻译：
拉曼氘同位素探测揭示了单细胞水平的微生物代谢
我们举例说明，单细胞拉曼光谱，结合氘同位素探测（Raman-DIP），提供了一种独立于培养且非破坏性的方法来探测单细胞水平上碳底物的代谢途径。当大肠杆菌使用氘代葡萄糖和假单胞菌时，我们在单细胞拉曼光谱（SCRS）中发现了2070–2300 cm –1处明显的C–D振动带。sp。使用氘代萘作为唯一碳源。C–D谱带的强度与碳源中氘的程度成正比，而通过SCRS分析可以分辨出只有5％的氘。这表明拉曼-DIP可用于半定量和灵敏地指示氘代碳源在微生物中的代谢。与氘化葡萄糖相比，氘化萘的脂质转化率较低，这可能是由于合成代谢途径和膜改变不同所致。除了从C–H到C–D的频移外，SCRS还揭示了苯丙氨酸带的几种同位素频移，其位置与计算模型很好地相关。减少了假单胞菌sp的苯丙氨酸氘化。相比于大肠杆菌归因于假单胞菌属物种中苯丙氨酸生物合成不同途径的稀释作用。集体地，我们证明拉曼-DIP不仅可以表明使用氘代碳源的代谢活性，而且还可以通过分析SCRS揭示不同的代谢途径。通过利用这种低成本，通用的氘代底物，拉曼DIP有望在单细胞水平上探索广泛的代谢途径和功能。
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