Skip to main content
SpringerLink
Log in

Partial purification and characterization of free and immobilized lipases fromMucor miehei

  • Published:
Lipids

An Erratum to this article was published on 01 September 1987

Abstract

An extracellular lipase, a glycoprotein, produced by fermentation with a selected strain ofMucor miehei has been partially purified in two forms, A and B. The two forms have a high degree of antigenic identity and have similar pH-activity profiles with tributyroylglycerol as a substrate with optima at pH 7. They differ as follows: A, in contrast to B, requires activation at alkaline pH before analysis; A binds with concanavalin-A more completely than B; the net charges are slightly different at pH 8; and the isoelectric points are different. Our results indicate that the B lipase is formed by partial deglycosylation of the A lipase and that this influences the activity toward emulsions.

In addition, the two enzymes have been immobilized by adsorption. These preparations and the soluble forms were highly specific for primary esters of triacylglycerols (TG); they usually hydrolyzed TG of 12∶0, 14∶0, 16∶0, and 18∶1 more rapidly than those of 4∶0, 6∶0, and 8∶0 and 10∶0 in mixtures of monoacid TG (4∶0 to 18∶1); and they were not stereospecific for TG. Immobilization altered the specificity of the preparations somewhat, in that slightly more 14∶0 and 16∶0 were released.

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

TG:

triacylglycerol

DG:

diacylglycerol

MG:

monoacylglycerol

conA:

concanavalin A

TLC:

thin layer chromatography

GLC:

gas liquid chromatography

LU:

lipase unit

NLU:

Nova lipase unit

pI:

isoelectric point

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

IEF:

isoelectric focusing

CIE:

crossed immunoelectrophoresis

CIAE:

crossed immunoaffinity electrophoresis

References

  1. Kilara, A. (1985)Process Biochem. 20, 36–45.

    Google Scholar 

  2. Moskowitz, G.J., Cassaigne, R., West, I.R., and Feldman, L.I. (1977)J. Agric. Food Chem. 25, 1146–1150.

    Article  PubMed  CAS  Google Scholar 

  3. Eigtved, P., Hansen, T.T., and Huge-Jensen, B. (1985)Proceedings of the 13th Scandinavian Symposium on Lipids (Lipidforum) pp. 135–148, Scandinavian Forum Lipid Res. Technol., Goteborg, Sweden.

    Google Scholar 

  4. Nielsen, T. (1985)Fette Seifen Anstrichm. 87, 15–19.

    Article  CAS  Google Scholar 

  5. Lavayre, J., and Baratti, J. (1982)Biotechnol. Bioeng. 24, 1007–1013.

    Article  CAS  PubMed  Google Scholar 

  6. Roland, J.F., Wargel, R.J., Alm, W.L., Kiang, S.P., and Bliss, F.M. (1984)Appl. Biochem. Biotechnol. 9, 15–26.

    CAS  Google Scholar 

  7. Brockman, H.L. (1981)Methods Enzymol. 71, 619–627.

    Article  PubMed  CAS  Google Scholar 

  8. Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. (1951)J. Biol. Chem. 193, 265–275.

    PubMed  CAS  Google Scholar 

  9. Bio-Rad Model 220 and 221,Vertical Slab Electrophoresis Cell Instruction Manual, April 1977.

  10. LKB manual 1804.Instructions. LKB Ampholine PAG Plates for Analytical Electrofucusing on Polyacrylamide Gels.

  11. Hojby, N., and Axelsen, N.H. (1983)Scand. J. Immunol. 17, Suppl. 10, 125–134.

    Google Scholar 

  12. Kroll, J. (1983)Scand. J. Immunol. 17, Suppl. 10, 135–139.

    Article  Google Scholar 

  13. Jensen, R.G., DeJong, F.A., and Clark, R.M. (1983)Lipids 18, 239–252.

    PubMed  CAS  Google Scholar 

  14. Jensen, R.G., Tardiff, G.S., and Clark, R.M. (1985)J. Pediatr. Gastroenterol. Nutr. 4, 580–582.

    Article  PubMed  CAS  Google Scholar 

  15. Wang, C.-S., Kuksis, A., and Managanaro, F. (1982)Lipids 17, 278–284.

    PubMed  CAS  Google Scholar 

  16. Jensen, R.G., DeJong, F.A., Clark, R.M., Palmgren, L.G., Lia, T.H., and Hamosh, M. (1982)Lipids 17, 570–572.

    PubMed  CAS  Google Scholar 

  17. Jensen, R.G., and Pitas, R.E. (1976)Adv. Lipid Res. 14, 213–247.

    CAS  Google Scholar 

  18. Wang, C-S., Kuksis, A., Manganaro, F., Myher, J.J., Downs, D., and Bass, H.B. (1983)J. Biol. Chem. 258, 9197–9202.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Novo Industri A/S, DK-2880, Bagsvaerd, Denmark

    Birgitte Huge-Jensen

  2. Department of Nutritional Sciences, University of Connecticut, 06268, Storrs, Connecticut

    Donna Rubano Galluzzo & Robert G. Jensen

Authors
  1. Birgitte Huge-Jensen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Donna Rubano Galluzzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Robert G. Jensen
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Scientific Contribution No. 1192, Storrs Agricultural Experiment Station, University of Connecticut, Storrs, Connecticut.

An erratum to this article is available at http://dx.doi.org/10.1007/BF02533951.

About this article

Cite this article

Huge-Jensen, B., Galluzzo, D.R. & Jensen, R.G. Partial purification and characterization of free and immobilized lipases fromMucor miehei . Lipids 22, 559–565 (1987). https://doi.org/10.1007/BF02537281

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02537281

Keywords

Search

Navigation