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Recent advances in the analysis of the site-specific isotopic fractionation of metabolites such as fatty acids using anisotropic natural-abundance 2H NMR spectroscopy: application to conjugated linolenic methyl esters

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Abstract

The full elucidation of the enzymatic mechanisms leading to polyunsaturated ω-3 to ω-5 fatty acids (PUFAs) occurring in plants or microorganisms by analyzing their site-specific isotopic fractionation profiles is a challenging task. Isotropic SNIF-NMR® method is an historical, powerful tool for the determination of (2H/1H) ratios. However, the absence of accessible isotopic data on the enantiotopic hydrogen sites (CH2 groups) prevents the study of the enzymatic reaction stereoselectivity. Natural-abundance deuterium (NAD) 2D NMR experiment using chiral liquid crystals (CLC) as solvent is a new tool in this field, overcoming this limitation. In this work, we have explored various possibilities for optimizing the enantio-discrimination properties of CLC by changing the nature of the polypeptide and/or increasing the polarity of the organic co-solvents. We report also the first applications of TMU as co-solvent for preparing enantio-discriminating, homogenous polypeptide mesophases. The various experimental NAD NMR results recorded at an optimal sample temperature are discussed and compared in terms of number of discriminated 2H sites and magnitude of spectral separation for different PUFAs such as the linoleic and linolenic acids. The comparison of all NMR results shows that optimal results are obtained when CLC mixtures made of poly-γ-benzyl-l-glutamate (PBLG) and high polarity co-solvents are used. As new challenging examples of applications, we report the preliminary analytical results obtained from two ω-5 conjugated linolenic acids: the α-eleostearic acid (9Z, 11E, 13E) and the punicic acid (9Z, 11E, 13Z). NMR data are discussed in terms of molecular orientational ordering parameters and isotopic distribution.

The new challenges of the site-specific isotopic fractionation analysis of fatty acids using the natural-abundance deuterium NMR in polypeptide aligned media

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References

  1. Martin GJ (1995) In: Wada E (ed) Stable isotopes in the biosphere. Kyoto University Press, Japan, pp 36–58

    Google Scholar 

  2. Duan J-R, Billault I, Mabon F, Robins RJ, Akoka S (2002) ChemBioChem 3:752–759

    Article  CAS  Google Scholar 

  3. Billault I, Guiet S, Mabon F, Robins RJ (2001) ChemBioChem 2:425–431

    Article  CAS  Google Scholar 

  4. Lesot P, Aroulanda C, Billault I (2004) Anal Chem 76:2827–2835

    Article  CAS  Google Scholar 

  5. Baillif V, Robins RJ, Billault I, Lesot P (2006) J Am Chem Soc 128:11180–11187

    Article  CAS  Google Scholar 

  6. Lesot P, Baillif V, Billault I (2008) Anal Chem 80:2963–2972

    Article  CAS  Google Scholar 

  7. Baillif V, Robins RJ, Le Feunteun, S, Lesot P, Billault I (2009) J Biol Chem 284:10783–110792

    Article  CAS  Google Scholar 

  8. Sarfati M, Lesot P, Merlet, D, Courtieu, J (2000) Chem Commun 2069-2081 and references therein

  9. Lesot P, Sarfati M, Courtieu J (2003) Chem Eur J 9:1724–1745

    Article  CAS  Google Scholar 

  10. Aroulanda C, Merlet D, Courtieu J, Lesot P (2001) P J Am Chem Soc 123:12059–12066

    Article  CAS  Google Scholar 

  11. Lafon O, Lesot P, Fan C-A, Kagan HB (2007) Chem Eur J 13:3772–3786

    Article  CAS  Google Scholar 

  12. Serhan Z, Martel L, Billault I, Lesot P (2010) Chem Commun 46:6599–6511

    Article  CAS  Google Scholar 

  13. Solgadi A, Jean L, Lasne M-C, Rouden J, Courtieu J, Meddour A (2007) Tetrahedron: Asymmetry 18:1511–1516

    Article  CAS  Google Scholar 

  14. Aroulanda C, Sarfati M, Courtieu J, Lesot P (2001) Enantiomer 6:281–287

    CAS  Google Scholar 

  15. Lesot P, Lafon O, Aroulanda C, Dong R (2008) Chem Eur J 14:4082–4092

    Article  CAS  Google Scholar 

  16. Harwood J (1997) In: Dey P, Harbone J (eds) Plant lipid metabolism in plant biochemistry. Academic, San Diego, pp 237–272

    Google Scholar 

  17. Ohlrogge J, Browse J (1995) Plant J 7:957–970

    CAS  Google Scholar 

  18. Scheim DE Lipids in Health and Disease, (Open access), (2009) 8:54 doi:101186/1476-511X-8-54 from http://www.lipidworld.com/content/8/1/54

  19. Grossmann ME, Mizuno NK, Schuster T, Margot P (2010) Int J Oncol 36:421–426

    Google Scholar 

  20. Lesot P, Courtieu J (2009) Prog Nucl Magn Reson Spect 55:128–159

    Google Scholar 

  21. The Handbook of Chemistry, Ed. 1977

  22. For TMU see: Riddick JA, Bunger WB, Sakano TK. In: Organic solvent, 4edn. Wiley Interscience NY, 1986

  23. Barwick VJ (1997) Trends Anal Chem 6:293–309

    Article  Google Scholar 

  24. The value is calculated for a single 2H site and the sites with the maximal number of equivalent 2H. For calculation the density of all components is assumed to be equal to 1.

  25. Marx A, Böttcher B, Thiele C (2010) Chem Eur J 16:1656–1663

    Article  CAS  Google Scholar 

  26. Kris-Etherton PM, Harris WS, Appel LJ, for the Nutrition Committee (2002) Fish consumption, fish oil, omega-3 fatty acids and cardiovascular diseases. Circulation 106:2747–2757

    Article  Google Scholar 

  27. Connor WE (2000) Am J Clin Nutr 71(1Suppl):171S–175S

    CAS  Google Scholar 

  28. Yasui Y, Hosokawa M, Sahara T, Suzuki R, Ohgiya S, Kohno H, Tanaka T, Miyashita K (2005) Prostaglandins Leukot Essent Fat Acids 73:113–119

    Article  CAS  Google Scholar 

  29. Dyer JM, Chapital DC, Kuan J-CW, Mullen RT, Turner C, McKeon TA, Pepperman AB (2002) Plant Physiol 130:2027–2038

    Article  CAS  Google Scholar 

  30. Liu L, Hammond EG, Nikolau BJ (1997) Plant Physiol 113:1343–1349

    CAS  Google Scholar 

  31. Crombie L, Holloway SJ (1985) J Chem Soc Perkin Trans 1:2425–2434

    Article  Google Scholar 

  32. Behrouzian B, Buist PH (2002) Curr Opin Chem Biol 6:577–582

    Article  CAS  Google Scholar 

  33. Reed DW, Savile CK, Qiu X, Buist PH, Covello PS (2002) Eur J Biochem 269:5024–5029

    Article  CAS  Google Scholar 

  34. Emsley JW, Lesot P, Courtieu J, Merlet D (2004) Phys Chem Chem Phys 6:5331–5337

    Article  CAS  Google Scholar 

  35. Zannoni C (1995) In: Nuclear magnetic resonance of liquid crystals. NATO ASI Series Reidel, Dordrecht, 1985, pp 1

  36. Emsley JW (1995) In: Encyclopedia of nuclear magnetic resonance. Wiley, Chichester, pp 2788

  37. Burnell EE, de Lange CA (2003) In: NMR of ordered liquids. Kluwer, Dordrecht, pp 5

  38. Aroulanda C, Lafon O, Lesot P (2009) J Phys Chem B 113:10628–10640

    Article  CAS  Google Scholar 

  39. Lafon O, Lesot P, Merlet D, Courtieu J. (2004) J Magn Reson 171: 135–142

    Google Scholar 

Download references

Acknowledgments

P. L. gratefully acknowledges Profs. M. Longeri and A. Loewenstein as well as Dr. P. Judeinstein for scientific discussions and/or their help during the preparation of this manuscript. The authors also thank CNRS for its financial support. Z. S. thanks the MNERS for her Ph.D granting.

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Authors and Affiliations

  1. Université de Paris Sud 11, CNRS UMR 8182 ICMMO, RMN en Milieu Orienté, Bât. 410, 91405, Orsay Cedex, France

    Philippe Lesot & Zeinab Serhan

  2. Université de Nantes, Elucidation of Biosynthesis by Isotopic Spectrometry Group, CEISAM, UMR CNRS 6230, CNRS UMR 6230, 2 rue de la Houssinière, BP 92208, 44322, Nantes Cedex, France

    Isabelle Billault

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  1. Philippe Lesot
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  2. Zeinab Serhan
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  3. Isabelle Billault
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Correspondence to Philippe Lesot.

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Lesot, P., Serhan, Z. & Billault, I. Recent advances in the analysis of the site-specific isotopic fractionation of metabolites such as fatty acids using anisotropic natural-abundance 2H NMR spectroscopy: application to conjugated linolenic methyl esters. Anal Bioanal Chem 399, 1187–1200 (2011). https://doi.org/10.1007/s00216-010-4394-0

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  • DOI: https://doi.org/10.1007/s00216-010-4394-0

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