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A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

Published on: June 1, 2017

Quantitative analysis of tetrahydrofolate metabolites from clostridium autoethanogenum

Renato de Souza Pinto Lemgruber¹, Kaspar Valgepea¹, Mark P Hodson^1,2

¹Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, 4072, Australia.

Metabolomics : Official Journal of the Metabolomic Society

|March 5, 2019

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.

Keywords:

Acetogens Gas fermentation Metabolome Tetrahydrofolates Wood–Ljungdahl pathway

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