Skip to main content
SpringerLink
Log in

Purification of plasmalogens usingRhizopus delemar lipase andNaja naja naja phospholipase A2

  • Methods
  • Published:
Lipids

Abstract

Bovine heart ChoGpl (choline glycerophospholipid) and bovine brain EtnGpl (ethanolamine glycerophospholipid) contain diacyl, alkenylacyl and alkylacyl analogs. Purification of plasmalogens was achieved usingR. delemar lipase andN. naja naja phospholipase A2 digestion. TheR. delemar lipase hydrolyzes the acyl bond at the 1-position of 1,2-diacyl glycerophospholipids. TheN. naja naja phospholipase A2 has greater activity with diacyl and alkylacyl than with alkenylacyl glycerophospholipids. These enzymes were mainly used to remove diacyl and alkylacyl analogs respectively. When the diacyl types were removed by double incubation withR. delemar lipase, the plasmalogen content was 94.2%±0.21% (mean±S.E.M., n=4) for PlsCho (plasmenylcholine) and 94.9%±0.19% (mean±S.E.M., n=3) for PlsEtn (plasmenylethanolamine). Recoveries were 74% and 88% respectively. These partially purified plasmalogens were treated withN. naja naja phospholipase A2. Finally, 97.7%±0.24% (mean±S.E.M., n=4) and 98.8%±0.27% (mean±S.E.M., n=3) pure plasmalogens were obtained for PlsCho and PlsEtn respectively. Plasmalogens were recovered in an overall yield of 7.7%±0.7% (mean±S.E.M., n=4) and 10.2%±1.2% (mean±S.E.M., n=3) for PlsCho and PlsEtn.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ChoGpl:

choline glycerophospholipid, 1,2-diradyl-sn-glycero-3-phosphocholine

EtnGpl:

ethanolamine glycerophospholipid, 1,2-diradyl-sn-glycero-3-phosphoethanolamine

PlsCho:

plasmenylcholine, 1-alk-1′-enyl-2-acyl-sn-glycero-3-phosphocholine

PlsEtn:

plasmenylethanolamine, 1-alk-1′-enyl-2-acyl-sn-glycero-3-phosphoethanolamine

PakCho:

phosphalkanylcholine, 1-alkyl-2-acyl-sn-glycero-3-phosphocholine

PakEtn:

phosphalkanylethanolamine, 1-alkyl-2-acyl-sn-glycero-3-phosphoethanolamine

References

  1. Horrocks, L.A., and Sharma, M. (1982) inPhospholipids, New Comprehensive Biochemistry (Hawthorne, J.N., and Ansell, G.B., eds.) Vol. 4, pp. 51–93, Elsevier Biomedical Press, Amsterdam.

    Google Scholar 

  2. Gottfried, E.L., and Rapport, M.M. (1962)J. Biol. Chem. 237, 329–333.

    PubMed  CAS  Google Scholar 

  3. Warner, H.R., and Lands, W.E.M. (1963)J. Am. Chem. Soc. 85, 60–64.

    Article  CAS  Google Scholar 

  4. Lands, W.E.M., and Hart, P. (1965)Biochim. Biophys. Acta 98, 532–538.

    PubMed  CAS  Google Scholar 

  5. Renkonen, O. (1963)Acta Chem. Scand. 17, 634–640.

    Article  CAS  Google Scholar 

  6. Frosolono, M.F., and Marsh, M. (1973)Chem. Phys. Lipids 10, 203–214.

    Article  PubMed  CAS  Google Scholar 

  7. Woelk, H., Debuch, H., and Porcellati, G. (1973)Hoppe-Seyler's Z. Physiol. Chem. 354, 1265–1270.

    PubMed  CAS  Google Scholar 

  8. De Haas, G.H., Sarda, L., and Roger, J. (1965)Biochim. Biophys. Acta 106, 638–640.

    PubMed  Google Scholar 

  9. Slotboom, A.J., de Haas, G.H., Bonsen, P.P.M., Burbach-Westerhuis, G.J., and van Deenen, L.L.M. (1970)Chem. Phys. Lipids 4, 15–29.

    Article  CAS  Google Scholar 

  10. Cox, J.W. (1977)Fed. Proc. 36, 852.

    Google Scholar 

  11. Paltauf, F. (1977)Lipids 13, 165–166.

    Article  Google Scholar 

  12. Pietruszko, R., and Gray, G.M. (1962)Biochim. Biophys. Acta 56, 232–239.

    Article  PubMed  CAS  Google Scholar 

  13. Pugh, E.L., Kates, M., and Hanahan, D.J. (1977)J. Lipid Res. 18, 710–716.

    PubMed  CAS  Google Scholar 

  14. Nakagawa, Y., and Horrocks, L.A. (1983)J. Lipid Res. 24, 1268–1275.

    PubMed  CAS  Google Scholar 

  15. Blank, M.L., Cress, E.A., and Snyder, F. (1987)J. Chromatogr. 392, 421–425.

    Article  PubMed  CAS  Google Scholar 

  16. Dembitsky, V.M. (1988)J. Chromatogr. 436, 467–473.

    Article  PubMed  CAS  Google Scholar 

  17. Waku, K., and Nakazawa, Y. (1972)J. Biochem. 72, 149–155.

    PubMed  CAS  Google Scholar 

  18. Sun, G.Y., deSousa, B.N., Danopoulos, V., and Horrocks, L.A. (1983)Int. J. Devl. Neurosci. 1, 59–64.

    Article  CAS  Google Scholar 

  19. Rouser, G., Fleischer, S., and Yamamoto, A. (1970)Lipids 5, 494–496.

    Article  PubMed  CAS  Google Scholar 

  20. Jones, M., Keenan, R.W., and Horowitz, P. (1982)J. Chromatogr. 237, 522–524.

    Article  CAS  Google Scholar 

  21. Horrocks, L.A., Yeo, Y.K., Harder, H.W., Mozzi, R., and Goracci, G. (1986)Adv. Cyclic Nucleotide Protein Phosphorylation Res. 20, 263–292.

    PubMed  CAS  Google Scholar 

  22. Hanahan, D.J., and Kumar, R. (1987)Prog. Lipid Res. 26, 1–28.

    Article  PubMed  CAS  Google Scholar 

  23. Freeman, N.M., and Carey, E.M. (1983)Anal. Biochem. 128, 377–383.

    Article  PubMed  CAS  Google Scholar 

  24. Arthur, G., Page, L., Mock, T., and Choy, P.C. (1986)Biochem. J. 236, 475–480.

    PubMed  CAS  Google Scholar 

  25. Hirashima, Y., Jurkowitz-Alexander, M.S., Farooqui, A.A., and Horrocks, L.A. (1989)Anal. Biochem. 176, 180–184.

    Article  PubMed  CAS  Google Scholar 

  26. Jurkowitz-Alexander, M., Ebata, H., Mills, J.S., Murphy, E.J., and Horrocks, L.A. (1989)Biochim. Biophys. Acta 1002 203–212.

    PubMed  CAS  Google Scholar 

  27. Hirashima, Y., Farooqui, A.A., and Horrocks, L.A. (1989)Biochem. J. 260, 605–608.

    PubMed  CAS  Google Scholar 

  28. Francescangeli, E., Porcellati, S., Horrocks, L.A., and Goracci, G. (1987)J. Liquid Chromatog. 10, 2799–2808.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiological Chemistry, The Ohio State University, 1645 Neil Avenue, 43210, Columbus, OH

    Yutaka Hirashima, Akhlaq A. Farooqui, Eric J. Murphy & Lloyd A. Horrocks

Authors
  1. Yutaka Hirashima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Akhlaq A. Farooqui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eric J. Murphy
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lloyd A. Horrocks
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Hirashima, Y., Farooqui, A.A., Murphy, E.J. et al. Purification of plasmalogens usingRhizopus delemar lipase andNaja naja naja phospholipase A2 . Lipids 25, 344–348 (1990). https://doi.org/10.1007/BF02544345

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02544345

Keywords

Search

Navigation