Skip to main content

HR MAS NMR Spectroscopy of Marine Microalgae

  • Reference work entry
  • First Online:

Abstract

Microalgae provide organic carbon through primary production and although most species are harmless, a few are toxic or otherwise damaging to animal and human life. Microalgae are used to produce biodiesel or feed in aquaculture or health food for human consumption, and species determination and knowledge of nutritional value is therefore needed. Common methods used in studies of microalgae provide only partial information, and sample preparation is often tedious and requires extraction with organic solvents. Application of high-resolution magic angle spinning (HR MAS)1H NMR spectroscopy gave well-resolved spectra directly from whole cells of several different marine microalgae, and comparison with spectra from extraction samples showed that both analyses provide to a large extent the same information on metabolite composition. The application of HR MAS NMR leads to increased signal resolution in semi-solid samples such as whole cells and organs, and this has made NMR a very interesting analytical tool to biologists, e.g., for metabolic profiling of microalgae.

Polyunsaturated fatty acids (PUFAs) eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids were seen in a 1H NMR spectrum of lipophilic extract of the diatom Thalassiosira pseudonana, while in a spectrum of hydrophilic extract we assigned signals to amino acids glutamine (Gln) and glutamic acid (Glu), carbohydrate, and ATP. Furthermore, HR MAS1H NMR spectroscopy in combination with the multivariate statistical principal component analysis (PCA) was used to separate groups of marine microalgae, and our results indicate that it is possible also to classify (or separate) species within the different groups. The PCA outcome was influenced by cellular content of various osmolytes, which indicates that the NMR spectra reflect signals from nuclei in liquid cellular compartments. This can be used to study cellular dynamics and transport of metabolites between different cellular locations, but also indicates that nuclei in other cellular locations are partly suppressed in the NMR analysis. Since the use of HR MAS1H NMR spectroscopy is relatively new in biological studies the methodological possibilities and restrains should be investigated closely, but our results support the use of NMR spectroscopy on whole-cell samples of microalgae.

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

Buying options

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   1,200.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD   1,299.99
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

Learn about institutional subscriptions

References

  1. Sarpal AS, Teixeira CM, Silva PR, da Costa Monteiro TV, da Silva JI, da Cunha VS, Daroda RJ. NMR techniques for determination of lipid content in microalgal biomass and their use in monitoring the cultivation with biodiesel potential. Appl Microbiol Biotechnol. 2016;100(5):2471–85.

    Article  CAS  Google Scholar 

  2. Brown MR, Dunstan GA, Norwood SJ, Miller KA. Effects of harvest stage and light on the biochemical composition of the diatom Thalassiosira pseudonana. J Phycol. 1996;32:64–73.

    Article  CAS  Google Scholar 

  3. McNamee SE, Medlin LK, Kegel J, McCoy GR, Raine R, Barra L, Ruggiero MV, Kooistra WHCF, Montresor M, Hagstrom J and others. Distribution, occurrence and biotoxin composition of the main shellfish toxin producing microalgae within European waters: a comparison of methods of analysis. Harmful Algae. 2016;55:112–20.

    Article  CAS  Google Scholar 

  4. Jeffrey SW, Mantoura RFC, Wright SW, editors. Phytoplankton pigments in oceanography: guidelines to modern methods. Paris: Unesco; 1997.

    Google Scholar 

  5. Mackey MD, Mackey DJ, Higgins HW, Wright SW. CHEMTAX – a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Mar Ecol Prog Ser. 1996;144:265–83.

    Article  CAS  Google Scholar 

  6. Sumner LW, Mendes P, Dixon RA. Plant metabolomics: large-scale phytochemistry in the functional genomics era. Phytochemistry. 2003;62:817–36.

    Article  CAS  Google Scholar 

  7. Meiboom S, Gill D. Modified spin-echo method for measuring nuclear relaxation times. Rev Sci Instrum. 1958;29:688–91.

    Article  CAS  Google Scholar 

  8. Schripsema J, Erkelens C, Verpoorte R. Intra- and extracellular carbohydrates in plant cell cultures investigated by 1H-NMR. Plant Cell Rep. 1991;9:527–30.

    Article  CAS  Google Scholar 

  9. Potts BC, Deese AJ, Stevens GJ, Reily MD, Robertson DG, Theiss J. NMR of biofluids and pattern recognition: assessing the impact of NMR parameters on the principal component analysis of urine from rat and mouse. J Pharm Biomed Anal. 2001;26:463–76.

    Article  CAS  Google Scholar 

  10. Chauton MS, Størseth TR, Johnsen G. High-resolution magic angle spinning 1H NMR analysis of whole cells of Thalassiosira pseudonana (Bacillariophyceae): broad range analysis of metabolic composition and nutritional value. J Appl Phycol. 2003;15(6):533–42.

    Article  Google Scholar 

  11. Chauton MS, Optun OI, Bathen TF, Volent Z, Gribbestad IS, Johnsen G. HR MAS1H NMR spectroscopy analysis of whole cells of marine microalgae. Mar Ecol Prog Ser. 2003;256:57–62.

    Article  CAS  Google Scholar 

  12. Chauton MS, Størseth TR, Krane J. HR MAS DEPT 13C NMR analysis of whole cells of Chaetoceros muelleri (Bacillariophyceae), and comparison with 13C NMR and DEPT13C NMR analysis of lipophilic extracts. J Phycol. 2004;40:611–8.

    Article  CAS  Google Scholar 

  13. Hedges JI, Baldock JA, Gélinas Y, Lee C, Peterson ML, Wakeham SG. The biochemical and elemental compositions of marine plankton: a NMR perspective. Mar Chem. 2002;78:47–63.

    Article  CAS  Google Scholar 

  14. Pollesello P, Toffanin R, Murano E, Paoletti S, Rizzo R, Kvam BJ. Lipid extracts from different algal species: 1H and13C-NMR spectroscopic studies as a new tool to screen differences in the composition of fatty acids, sterols and carotenoids. J Appl Phycol. 1992;4(4):315–22.

    Article  CAS  Google Scholar 

  15. Størseth TR, Hansen K, Skjermo J, Krane J. Characterization of a β-D-(1 → 3)-glucan from the marine diatom Chaetoceros mülleri by high-resolution magic-angle spinning NMR spectroscopy on whole algal cells. Carbohydr Res. 2004;339:421–4.

    Article  CAS  Google Scholar 

  16. Davey PT, Hiscox WC, Lucker BF, O’Fallon JV, Chen SL, Helms GL. Rapid triacylglyceride detection and quantification in live micro-algal cultures via liquid state H-1 NMR. Algal Res-Biomass Biofuels Bioprod. 2012;1(2):166–75.

    Google Scholar 

  17. Arnold AA, Genard B, Zito F, Tremblay R, Warschawski DE, Marcotte I. Identification of lipid and saccharide constituents of whole microalgal cells by 13C solid-state NMR. Biochim Biophys Acta Biomembr. 2015;1848(1, Part B):369–77.

    Article  CAS  Google Scholar 

  18. Merkley N, Syvitski RT. Profiling whole microalgal cells by high-resolution magic angle spinning (HR-MAS) magnetic resonance spectroscopy. J Appl Phycol. 2012;24(3):535–40.

    Article  CAS  Google Scholar 

  19. Zea Obando C, Linossier I, Kervarec N, Zubia M, Turquet J, Faÿ F, Rehel K. Rapid identification of osmolytes in tropical microalgae and cyanobacteria by 1H HR-MAS NMR spectroscopy. Talanta. 2016;153:372–80.

    Article  CAS  Google Scholar 

  20. Gao CF, Xiong W, Zhang YL, Yuan WQ, Wu QY. Rapid quantitation of lipid in microalgae by time-domain nuclear magnetic resonance. J Microbiol Methods. 2008;75(3):437–40.

    Article  CAS  Google Scholar 

  21. Nuzzo G, Gallo C, D’Ippolito G, Cutignano A, Sardo A, Fontana A. Composition and quantitation of microalgal lipids by ERETIC H-1 NMR method. Mar Drugs. 2013;11(10):3742–53.

    Article  CAS  Google Scholar 

  22. Turpin DH, Harrison PJ. Fluctuations in free amino acid pools of Gymnodinium simplex (Dinophyceae) in response to ammonia perturbation: evidence for glutamine synthetase pathway. J Phycol. 1978;14:461–4.

    Article  Google Scholar 

  23. Derrien A, Coiffard LJM, Coiffard C, De Roeck-Holtzhauer Y. Free amino acid analysis of five microalgae. J Appl Phycol. 1998;10:131–4.

    Article  CAS  Google Scholar 

  24. Granum E. Metabolism and function of β-1,3-glucan in marine diatoms. PhD thesis, NTNU, Trondheim; 2002.

    Google Scholar 

  25. Nicholson JK, Foxall PJD. 750 MHz1H and1H-13C NMR spectroscopy of human blood plasma. Anal Chem. 1995;67:793–811.

    Article  CAS  Google Scholar 

  26. Sitter B, Sonnewald U, Spraul M, Fjosne HE, Gribbestad IS. High-resolution magic angle spinning MRS of breast cancer tissue. NMR Biomed. 2002;15(5):327–37.

    Article  CAS  Google Scholar 

  27. Willker W, Leibfritz D. Assignment of mono- and polyunsaturated fatty acids in lipids of tissues and body fluids. Magn Reson Chem. 1998;36(S1):S79–84.

    Article  CAS  Google Scholar 

  28. Lindon JC, Nicholson JK, Everett JR. NMR spectroscopy of biofluids. Ann Rep NMR Spectrosc. 1999;38:1–88.

    Article  CAS  Google Scholar 

  29. Ward JL, Harris C, Lewis J, Beale MH. Assessment of 1H NMR spectroscopy and multivariate analysis as a technique for metabolite fingerprinting of Arabidopsis thaliana. Phytochemistry. 2003;62:949–57.

    Article  CAS  Google Scholar 

  30. Chatham JC, Forder JR. Lactic acid and protein interactions: implications for the NMR visibility of lactate in biological systems. Biochim Biophys Acta. 1999;1426:177–84.

    Article  CAS  Google Scholar 

  31. Babuska R, Alic L, Lourens MS, Verbraak AFM, Bogaard J. Estimation of respiratory parameters via fuzzy clustering. Artif Intell Med. 2001;21:91–105.

    Article  CAS  Google Scholar 

  32. Martens H, Næs T. Multivariate calibration. New York: John Wiley & Sons; 1991.

    Google Scholar 

  33. Keller MD, Kiene RP, Matrai PA, Bellows WK. Production of glycine betaine and dimethylsulfoniopropionate in marine phytoplankton. II. N-limited chemostat cultures. Mar Biol. 1999;135:249–57.

    Article  CAS  Google Scholar 

  34. Bundy JG, Spurgeon DJ, Svendsen C, Hankard PK, Osborn D, Lindon JC, Nicholson JK. Earthworm species of the genus Eisenia can be phenotypically differentiated by metabolic profiling. FEBS Lett. 2002;521:115–20.

    Article  CAS  Google Scholar 

  35. Defernez M, Colquhoun IJ. Factors affecting the robustness of metabolite fingerprinting using1H NMR spectra. Phytochemistry. 2003;62:1009–17.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Matilde Skogen Chauton or Trond Størseth .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer International Publishing AG, part of Springer Nature

About this entry

Check for updates. Verify currency and authenticity via CrossMark

Cite this entry

Chauton, M.S., Størseth, T. (2018). HR MAS NMR Spectroscopy of Marine Microalgae. In: Webb, G. (eds) Modern Magnetic Resonance. Springer, Cham. https://doi.org/10.1007/978-3-319-28388-3_82

Download citation

Publish with us

Policies and ethics