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Smith, T.W. and Hippler, M.F. orcid.org/0000-0002-3956-3922 (2017) Cavity-Enhanced Raman Spectroscopy in the Biosciences: In situ, Multicomponent and Isotope Selective Gas Measurements to Study Hydrogen Production and Consumption by Escherichia coli. Analytical Chemistry. ISSN 1520-6882
Abstract
Recently we introduced Cavity-Enhanced Raman Spectroscopy (CERS) with optical feedback cw-diode lasers as a sensitive analytical tool. Here we report improvements made on the technique and its first application in the biosciences for in situ, multicomponent and isotope selective gas measurements to study hydrogen production and consumption by Escherichia coli. Under anaerobic conditions, cultures grown on rich media supplemented with D-glucose or glycerol produce H2 and simultaneously consume some of it. By introducing D2 in the headspace, hydrogen production and consumption could be separated due to the distinct spectroscopic signatures of isotopomers. Different phases with distinctly different kinetic regimes of H2 and CO2 production and D2 consumption were identified. Some of the D2 consumed is converted back to H2 via H/D exchange with the solvent. HD was formed only as a minor component. This reflects either that H/D exchange at hydrogenase active sites is rapid compared to the rate of recombination, rapid recapture of HD occurs after the molecule is formed, or that the active sites where D2 oxidation and proton reduction occur are physically separated. Whereas in glucose supplemented cultures, addition of D2 led to an increase in H2 produced while the yield of CO2 remained unchanged, with glycerol, addition of D2 led not only to increased yields of H2 but also significantly increased CO2 production, reflecting an impact on fermentation pathways. Addition of CO was found to completely inhibit H2 production and significantly reduce D2 oxidation, indicating at least some role for O2-tolerant Hyd-1 in D2 consumption.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2016 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) |
Funding Information: | Funder Grant number NATURAL ENVIRONMENT RESEARCH COUNCIL NE/I000844/1 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 19 Jan 2017 12:29 |
Last Modified: | 10 Jan 2018 01:38 |
Published Version: | https://doi.org/10.1021/acs.analchem.6b04924 |
Status: | Published |
Publisher: | American Chemical Society |
Refereed: | Yes |
Identification Number: | 10.1021/acs.analchem.6b04924 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:110719 |
Available Versions of this Item
- Cavity-Enhanced Raman Spectroscopy in the Biosciences: In situ, Multicomponent and Isotope Selective Gas Measurements to Study Hydrogen Production and Consumption by Escherichia coli. (deposited 19 Jan 2017 12:29) [Currently Displayed]