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Journal of Applied Phycology

, Volume 24, Issue 3, pp 535–540 | Cite as

Profiling whole microalgal cells by high-resolution magic angle spinning (HR-MAS) magnetic resonance spectroscopy

  • Nadine MerkleyEmail author
  • Raymond T. Syvitski
Article

Abstract

Microalgae are a rich source of high value compounds such as carbohydrates, lipids, proteins and bioactive compounds. In particular, microalgae have been identified as a potentially important resource for carbon-capture and as a feedstock for green biofuels. Successful cultivation of microalgae can occur under a variety of nutrient and environmental conditions with each condition producing a unique distribution of compounds. In order to steer the cultivation towards a particular distribution of compounds, rapid and accurate methods for compound identification are required. Current methods for determining the absolute quantity of each component are time consuming and arduous making cultivation optimization impractical. High-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy offers a robust and rapid screening method capable of ascertaining the absolute quantity of each component with minimal sample manipulation. Sample preparation consists of harvested, centrifuged and freeze-dried whole-cell Nannochloropsis granulata from large-scale photobioreactors being accurately weighed and rehydrated with deuterium oxide and placed in an HR-MAS rotor. One-dimensional HR-MAS NMR spectra were recorded under quantitative conditions to determine the lipid and carbohydrate profile of the microalgae. The total time per sample for preparation, data acquisition and analysis was approximately 1 h. Changes in resonance profiles corresponding to varying proportions of saturated and polyunsaturated fatty acids were correlated to the time of harvest. In addition, standard two dimensional experiments were used to identify the major carbohydrate components. HR-MAS NMR spectroscopy has been used to profile the lipid and carbohydrate content of N. granulata and we have begun to establish methodologies for quality analysis/quality control for cultivation of various microalgal strains.

Keywords

High-resolution magic angle spinning Nannochloropsis granulata Nuclear magnetic resonance spectroscopy Whole-cell lipid profiling Quality assurance/quality control 

Notes

Acknowledgements

Samples of N. granulata were obtained from Pat McGinn and Jenny MacPherson of the Institute for Marine Biosciences Marine Research Station. The authors thank Stephen O’Leary, Pat McGinn and Katie Dickinson for valuable discussions. Magnetic Resonance data was collected at the Biomolecular Magnetic Resonance Facility (BMRF) housed at the National Research Council of Canada's Institute for Marine Biosciences. The He-cooled probe for the 700 MHz NMR spectrometer at the NRC-IMB was provided by Dalhousie University through an Atlantic Canada Opportunities Agency Grant. This is NRC publication number 2011–54066.

References

  1. Bondu S, Kervarec N, Deslandes E, Pichon R (2008) The use of HRMAS NMR spectroscopy to study the in vivo intra-cellular carbon/nitrogen ratio of Solieria chordalis (Rhodophyta). J Appl Phycol 20:673–679CrossRefGoogle Scholar
  2. Broberg A, Kenne L, Pedersén M (1998) In situ identification of major metabolites in the red alga Gracilariopsis lemaneiformis using high-resolution magic angle spinning nuclear magnetic resonance spectroscopy. Planta 206:300–307CrossRefGoogle Scholar
  3. Brown MR (1991) The amino-acid and sugar composition of 16 species of microalgae used in mariculture. J Exp Mar Biol Ecol 145:79–99CrossRefGoogle Scholar
  4. Burton IW, Quilliam MA, Walter JA (2005) Quantitative 1H NMR with external standards: use in preparation of calibration solutions for algal toxins and other natural products. Anal Chem 77:3123–3131PubMedCrossRefGoogle Scholar
  5. Chauton MS, Optun OI, Bathen TF, Volent Z, Gribbestad IS, Johnsen G (2003a) HR MAS 1H NMR spectroscopy analysis of microalgal whole cells. Mar Ecol Prog Ser 256:57–62CrossRefGoogle Scholar
  6. Chauton MS, Størseth TR, Johnsen G (2003b) 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 15:533–542CrossRefGoogle Scholar
  7. Chauton MS, Størseth TR, Krane J (2004) High-resolution magic angle spinning NMR analysis of Chaetoceros muelleri (Bacillariophyceae) and comparison with 13 C-NMR and distortionless enhancement by polarization transfer 13 C-NMR analysis of lipophilic extracts. J Phycol 40:611–618CrossRefGoogle Scholar
  8. Chen W, Zhang C, Song L, Sommerfield M, Hu Q (2009) A high throughput Nile Red method for quantitative measurement of neutral lipids in microalgae. J Microbiol Methods 77:41–47PubMedCrossRefGoogle Scholar
  9. Cooksey KE, Guckert JB, Williams SA, Callis PR (1987) Fluorometric determination of the neutral lipid content of microalgal cells using Nile Red. J Microbiol Methods 6:333–345CrossRefGoogle Scholar
  10. Dittami SM, Aas HTN, Paulsen BS, Boyen C, Edvardsen B, Tonon T (2011) Mannitol in six autotrophic stramenopiles and Micromonas. Plant Signal Behav 6:1237–1239PubMedCrossRefGoogle Scholar
  11. Dunsten GA, Volkman JK, Barret SM, Garland CD (1993) Changes in the lipid composition and maximisation of the polyunsaturated fatty acid content of three microalgae grown in mass culture. J Appl Phycol 5:71–83CrossRefGoogle Scholar
  12. Guillard RRL (1975) Culture of phytoplankton for feeding marine invertebrates. In: Smith WL, Chanley MH (eds) Culture of marine invertebrate animals. Plenum, New York, pp 26–60Google Scholar
  13. Hara A, Radin NS (1978) Lipid extraction of tissues with a low-toxicity solvent. Anal Biochem 90:420–426PubMedCrossRefGoogle Scholar
  14. Hanoulle X, Wieruszeski J-M, Rousselot-Pailley P, Landrieu I, Baulard AR, Lippens G (2005) Monitoring of the ethionamide pro-drug activation in mycobacteria by 1H high resolution magic angle spinning NMR. Biochem Biophys Res Commun 331:452–458PubMedCrossRefGoogle Scholar
  15. Koskela H (2009) Quantitative 2D NMR studies. Annu Rep NMR Spectrosc 66:1CrossRefGoogle Scholar
  16. Laurens LML, Wolfrum EJ (2011) Feasibility of spectroscopic characterization of algal lipids: chemometric correlation of NIR and FTIR spectra with exogenous lipids in algal biomass. Bioenerg Res 4:22–35CrossRefGoogle Scholar
  17. Li W (2006) Multidimensional HRMAS NMR: a platform for in vivo studies using intact bacterial cells. Analyst 131:777–781PubMedCrossRefGoogle Scholar
  18. Lindon JC, Beckonert OP, Holmes E, Nicholson JK (2009) High-resolution magic angle spinning NMR spectroscopy: application to biomedical studies. Prog NMR Spectrosc 5:79–100CrossRefGoogle Scholar
  19. Nestor G, Bankefors J, Schlechtriem C, Brännäs E, Pickova J, Sandström C (2010) High-resolution 1H magic angle spinning NMR spectroscopy of intact Arctic Char (Salvelinus alpinus) muscle. Quantitative analysis of n-3 fatty acids, EPA and DHA. J Agric Food Chem 58:10799–10803CrossRefGoogle Scholar
  20. Power WP (2003) High resolution magic angle spinning – Applications to solid phase synthetic systems and other semi-solids. Annu Rep NMR Spectrosc 51:241–248Google Scholar
  21. Weybright P, Millis K, Campbell N, Cory DG, Singer S (1998) Gradient, high-resolution, magic angle spinning 1H nuclear magnetic resonance spectroscopy of intact cells. Magn Reson Med 39:337–344PubMedCrossRefGoogle Scholar

Copyright information

© Her Majesty the Queen in Right of Canada as represented by the National Research Council of Canada 2011

Authors and Affiliations

  1. 1.Institute for Marine Biosciences, National Research Council of CanadaHalifaxCanada

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