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Stable Isotope Probing of Microbial Phospholipid Fatty Acids in Environmental Samples

  • Andrea WatzingerEmail author
  • Rebecca Hood-Nowotny
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2046)

Abstract

Phospholipid fatty acid (PLFA) extracted from environmental samples describe the microbial community pattern and are sensitive to monitor and quantify shifts in the microbial community. Linkage with the stable isotope technique adds a functional perspective and is frequently used to quantify carbon turnover in microbial communities and detect physiological changes. Here we present a PLFA extraction method by using an organic solvent water mixture, followed by lipid separation based on solid-phase extraction and an alkaline methylation. Finally, we provide a protocol for the carbon stable isotope measurements of the extracted fatty acid methyl esters (FAMEs) by gas chromatograph–isotope ratio mass spectrometer (GC-IRMS) and calculation of concentration and δ13CVPDB values.

Key words

Phospholipid fatty acid Stable isotope probing Lipid extraction Separation Solid-phase extraction Alkaline methylation Fatty acid methyl ester Microorganisms Environmental samples GC-IRMS 

Notes

Acknowledgments

The authors would like to thank all the coworkers and students for critical questions and comments when conducting PLFA extraction and GC-IRMS measurements, which over the years improved the protocol. The County of Lower Austria, NÖ Wirtschafts- und Tourismusfonds (WST3-F-5031245/003-2018) is acknowledged for funding.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Soil ResearchUniversity of Natural Resources and Life Sciences ViennaTullnAustria

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