Renato de Souza Pinto Lemgruber1, Kaspar Valgepea1, Mark P Hodson1,2, Ryan Tappel3, Sean D Simpson3, Michael Köpke3, Lars K Nielsen1, Esteban Marcellin4,5. 1. Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, 4072, Australia. 2. Metabolomics Australia, AIBN, The University of Queensland, Brisbane, Australia. 3. LanzaTech Inc., Skokie, IL, 60077, USA. 4. Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, 4072, Australia. e.marcellin@uq.edu.au. 5. Metabolomics Australia, AIBN, The University of Queensland, Brisbane, Australia. e.marcellin@uq.edu.au.
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.
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.
Entities:
Keywords:
Acetogens; Gas fermentation; Metabolome; Tetrahydrofolates; Wood–Ljungdahl pathway
Authors: Lee C Garratt; Catharine A Ortori; Gregory A Tucker; Fred Sablitzky; Malcolm J Bennett; David A Barrett Journal: Rapid Commun Mass Spectrom Date: 2005 Impact factor: 2.419