Xu, J., Zhu, D., Ibrahim, A.D. et al. (5 more authors) (2017) Raman Deuterium Isotope Probing Reveals Microbial Metabolism at the Single-Cell Level. Analytical Chemistry, 89 (24). pp. 13305-13312. ISSN 0003-2700
Abstract
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2017 American Chemical Society. This is an author produced version of a paper subsequently published in Analytical Chemistry. Uploaded in accordance with the publisher's self-archiving policy. |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > Department of Chemistry (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 20 Dec 2017 13:30 |
Last Modified: | 14 Jan 2020 14:33 |
Published Version: | https://doi.org/10.1021/acs.analchem.7b03461 |
Status: | Published |
Publisher: | American Chemical Society |
Refereed: | Yes |
Identification Number: | 10.1021/acs.analchem.7b03461 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:125344 |