Skip to main content

Isolation and Analysis of Membrane Phospholipids as Biomarkers

  • Protocol
  • First Online:
Hydrocarbon and Lipid Microbiology Protocols

Part of the book series: Springer Protocols Handbooks ((SPH))

  • 642 Accesses

Abstract

The protocol describes the analysis and data interpretation of membrane phospholipid fatty acid (PLFA) profiles as biomarkers for the characterisation of microbial communities in environmental samples, with special emphasis on soils and sediments contaminated with organic hydrocarbons. This simple method in microbial ecology studies provides an estimate of the viable microbial biomass, first indications of the structure of the microbial community, as well as the physiological status of the microbiota. In environmental biotechnological applications, PLFA analysis is a useful tool for monitoring bioremediation and allows direct validation of existing biological activities as well as hints for shifts in the microbiota caused by successful biological treatment strategies.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Frostegard A, Baath E, Tunlid A (1993) Shifts in the structure of soil microbial communities in limed forests as revealed by phospholipid fatty-acid analysis. Soil Biol Biochem 25:723–730

    Article  Google Scholar 

  2. Frostegard A, Tunlid A, Baath E (1996) Changes in microbial community structure during long-term incubation in two soils experimentally contaminated with metals. Soil Biol Biochem 28:55–63

    Article  CAS  Google Scholar 

  3. Amann RI, Ludwig W, Schleifer KH (1995) Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 59:143–169

    CAS  PubMed Central  Google Scholar 

  4. Frostegard A, Tunlid A, Baath E (1993) Phospholipid fatty acid composition, biomass, and activity of microbial communities from 2 soil types experimentally exposed to different heavy metals. Appl Environ Microbiol 59:3605–3617

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Frostegård Å, Tunlid A, Bååth E (2011) Use and misuse of PLFA measurements in soils. Soil Biol Biochem 43:1621–1625

    Article  Google Scholar 

  6. MacNaughton SJ, Stephen JR, Venosa AD, Davis GA, Chang YJ, White DC (1999) Microbial population changes during bioremediation of an experimental oil spill. Appl Environ Microbiol 65:3566–3574

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Villanueva L, Navarrete A, Urmeneta J, White DC, Guerrero R (2004) Combined phospholipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis of bacterial diversity and physiological status in an intertidal microbial mat. Appl Environ Microbiol 70:6920–6926

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. White DC, Stair JO, Ringelberg DB (1996) Quantitative comparisons of in situ microbial biodiversity by signature biomarker analysis. J Ind Microbiol 17:185–196

    Article  CAS  Google Scholar 

  9. Zelles L (1997) Phospholipid fatty acid profiles in selected members of soil microbial communities. Chemosphere 35:275–294

    Article  CAS  PubMed  Google Scholar 

  10. Pennanen T, Frostegard A, Fritze H, Baath E (1996) Phospholipid fatty acid composition and heavy metal tolerance of soil microbial communities along two heavy metal-polluted gradients in coniferous forests. Appl Environ Microbiol 62:420–428

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Quezada M, Buitrón G, Moreno-Andrade MIG, López-Marín LM (2007) The use of fatty acid methyl esters as biomarkers to determine aerobic, facultatively aerobic and anaerobic communities in wastewater treatment systems. FEMS Microbiol Lett 266:75–82

    Article  CAS  PubMed  Google Scholar 

  12. Syakti AD, Mazzella N, Nerini D, Guiliano M, Bertrand JC, Doumenq P (2006) Phospholipid fatty acids of a marine sedimentary microbial community in a laboratory microcosm: responses to petroleum hydrocarbon contamination. Org Geochem 37:1617–1628

    Article  CAS  Google Scholar 

  13. White DC (1993) In situ measurement of microbial biomass, community structure and nutritional-status. Phil Trans R Soc Lond A 344:59–67

    Article  CAS  Google Scholar 

  14. Kaur A, Chaudhary A, Kaur A, Choudhary R, Kaushik R (2005) Phospholipid fatty acid - a bioindicator of environment monitoring and assessment in soil ecosystem. Curr Sci 89:1103–1112

    CAS  Google Scholar 

  15. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917

    Article  CAS  PubMed  Google Scholar 

  16. Morrison WR, Smith LM (1964) Preparation of fatty acid methyl esters and dimethylacetals from lipids with boron fluoride methanol. J Lipid Res 5:600–608

    CAS  PubMed  Google Scholar 

  17. Piotrowska-Seget Z, Mrozik A (2003) Signature lipid biomarker (SLB) analysis in determining changes in community structure of soil microorganisms. Polish J Environ Stud 12:669–675

    CAS  Google Scholar 

  18. Kieft TL, Ringelberg DB, White DC (1994) Changes in ester-linked phospholipid fatty acid profiles of subsurface bacteria during starvation and desiccation in a porous-medium. Appl Environ Microbiol 60:3292–3299

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Kabelitz N et al (2009) Enhancement of the microbial community biomass and diversity during air sparging bioremediation of a soil highly contaminated with kerosene and BTEX. Appl Microbiol Biotechnol 82:565–577

    Article  CAS  PubMed  Google Scholar 

  20. de Carvalho CCCR, Marques M, Hachicho N, Heipieper H (2014) Rapid adaptation of Rhodococcus erythropolis cells to salt stress by synthesizing polyunsaturated fatty acids. Appl Microbiol Biotechnol 98:5599–5606

    PubMed  Google Scholar 

  21. Keweloh H, Heipieper HJ (1996) Trans unsaturated fatty acids in bacteria. Lipids 31:129–137

    Article  CAS  PubMed  Google Scholar 

  22. Unell M, Kabelitz N, Jansson JK, Heipieper HJ (2006) Adaptation of the psychrotroph Arthrobacter chlorophenolicus A6 to growth temperature and the presence of phenols by changes in the anteiso/iso ratio of branched fatty acids. FEMS Microbiol Lett 266:138–143

    Article  Google Scholar 

  23. Margesin R, Hammerle M, Tscherko D (2007) Microbial activity and community composition during bioremediation of diesel-oil-contaminated soil: effects of hydrocarbon concentration, fertilizers, and incubation time. Microb Ecol 53:259–269

    Article  CAS  PubMed  Google Scholar 

  24. Vestal JR, White DC (1989) Lipid analysis in microbial ecology: quantitative approaches to the study of microbial communities. Bioscience 39:535–541

    Article  CAS  PubMed  Google Scholar 

  25. Ramos JL, Gallegos MT, Marques S, Ramos-Gonzalez MI, Espinosa-Urgel M, Segura A (2001) Responses of Gram-negative bacteria to certain environmental stressors. Curr Opin Microbiol 4:166–171

    Article  CAS  PubMed  Google Scholar 

  26. Heipieper HJ, Meinhardt F, Segura A (2003) The cis-trans isomerase of unsaturated fatty acids in Pseudomonas and Vibrio: biochemistry, molecular biology and physiological function of a unique stress adaptive mechanism. FEMS Microbiol Lett 229:1–7

    Article  CAS  PubMed  Google Scholar 

  27. Kool DM, Zhu BL, Rijpstra WIC, Jetten MSM, Ettwig KF, Damste JSS (2012) Rare branched fatty acids characterize the lipid composition of the intra-aerobic methane oxidizer “Candidatus Methylomirabilis oxyfera”. Appl Environ Microbiol 78:8650–8656

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Londry KL, Jahnke LL, Marais DJD (2004) Stable carbon isotope ratios of lipid biomarkers of sulfate-reducing bacteria. Appl Environ Microbiol 70:745–751

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Lerch TZ, Dignac MF, Barriuso E, Mariotti A (2011) Effect of glucose on the fatty acid composition of Cupriavidus necator jmp134 during 2,4-dichlorophenoxyacetic acid degradation: implications for lipid-based stable isotope probing methods. Appl Environ Microbiol 77:7296–7306

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Yan X, Gurtler J, Fratamico P, Hu J, Gunther NW, Juneja V, Huang L (2011) comprehensive approaches to molecular biomarker discovery for detection and identification of Cronobacter spp. (Enterobacter sakazakii) and Salmonella spp. Appl Environ Microbiol 77:1833–1843

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hermann J. Heipieper .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer-Verlag Berlin Heidelberg

About this protocol

Cite this protocol

Hachicho, N., Heipieper, H.J. (2014). Isolation and Analysis of Membrane Phospholipids as Biomarkers. In: McGenity, T., Timmis, K., Nogales , B. (eds) Hydrocarbon and Lipid Microbiology Protocols . Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8623_2014_17

Download citation

  • DOI: https://doi.org/10.1007/8623_2014_17

  • Published:

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-49135-5

  • Online ISBN: 978-3-662-49137-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics