Abstract
Introduction
Quantification of tetrahydrofolates (THFs), important metabolites in the Wood–Ljungdahl pathway (WLP) of acetogens, is challenging given their sensitivity to oxygen.
Objective
To develop a simple anaerobic protocol to enable reliable THFs quantification from bioreactors.
Methods
Anaerobic cultures were mixed with anaerobic acetonitrile for extraction. Targeted LC–MS/MS was used for quantification.
Results
Tetrahydrofolates can only be quantified if sampled anaerobically. THF levels showed a strong correlation to acetyl-CoA, the end product of the WLP.
Conclusion
Our method is useful for relative quantification of THFs across different growth conditions. Absolute quantification of THFs requires the use of labelled standards.
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Acknowledgements
This study was funded by a Grant from the Australian Research Council, partly funded by LanzaTech (ARC LP140100213). Elements of this research utilised equipment and support provided by the QLD node of Metabolomics Australia, an initiative of the Australian Government being conducted as part of the NCRIS National Research Infrastructure for Australia.
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RSPL, KV, MPH, LKN, EM declare that they have no conflict of interest. LanzaTech has interest in commercial gas fermentation with C. autoethanogenum. RT, SDS, MK are employees of LanzaTech.
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de Souza Pinto Lemgruber, R., Valgepea, K., Hodson, M.P. et al. Quantitative analysis of tetrahydrofolate metabolites from clostridium autoethanogenum. Metabolomics 14, 35 (2018). https://doi.org/10.1007/s11306-018-1331-2
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DOI: https://doi.org/10.1007/s11306-018-1331-2