Advertisement

Biotechnology Letters

, Volume 26, Issue 11, pp 933–937 | Cite as

Flow process for electroextraction of intracellular enzymes from the fission yeast, Schizosaccharomyces pombe

  • V. Ganeva
  • B. Galutzov
  • J. Teissié
Article

Abstract

Flow treatment of the yeast, Schizosaccharomyces pombe, with high intensity electric field pulses released intracellular enzymes such as glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase. Over 70% of the total activity was liberated within 4 h after pulse application. The optimal field intensities were considerably higher than that needed for irreversible plasma membrane permeabilization.

cytoplasmic enzyme electro extraction electropulsation flow process Schizosaccharomyces pombe 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. De Nobel JG, Klis FM, Munnik T, Priem J, Van den Ende H (1990) An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. Yeast 6: 483–490.Google Scholar
  2. Ganeva V, Galutzov B (1999) Electropulsation as an alternative method for protein extraction from yeast. FEMS Lett. 174: 279–284.Google Scholar
  3. Ganeva V, Galutzov B, Teissie J (1995) Electric field mediated loading of macromolecules in in intact years cells is critically controlled at the wall level. Biochim. Biophys. Acta 1240: 329–336.Google Scholar
  4. Ganeva V, Galutzov B, Teissie J (2002) Electroinduced release of invertase from Saccharomyces cerevisiae. Biotechnol. Lett. 24: 1853–1856.Google Scholar
  5. Ganeva V, Galutzov B, Teissie J (2003) High yield electroextraction of proteins from yeast by flow process. Anal. Biochem. 315: 77–84.Google Scholar
  6. Kirschner K, Voigt B (1968) Purity criteria for the crystallizable isoenzyme of D-glyceraldehyde-3-phosphate dehydrogenase from baker's yeast. Hoppe Seyler's Z. Physiol. Chem. 349:632–643.Google Scholar
  7. Kopecka M (1995) Ultrastructure of the cell wall of Schizosaccharomyces pombe following treatment with various glucanases. J. Struct. Biol. 114: 140–152.Google Scholar
  8. Kulbe K, Bojanovski M (1982) 3-Phosphoglycerate kinase from bovine liver and yeast. Meth. Enzymol. 90: 115–120.Google Scholar
  9. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 22: 680–685.Google Scholar
  10. Nesterenko MV, Tilley M, Upton SJ (1994) A simple modification of Blum's silver stain method allows for 30 min detection of proteins in polyacrylamide gels. J. Biochem. Biophys. Meth. 28: 239–242.Google Scholar
  11. Neumann E, Sprafke A, Boldt E, Wolf H (1992) Biophysical consideration of membrane electroporation. In: Chang DC, Chassy BM, eds. Guide to Electroporation and Electrofusion. NewYork: Academic Press, pp. 77–91.Google Scholar
  12. Rols MP, Teissie J (1990) Electropermeabilization of mammalian cells. Quantitative analysis of the phenomenon. Biophys. J. 58: 1089–1098.Google Scholar
  13. Zlotnik H, Fernandez MP, Bowers B, Cabib E (1984) Saccharomyces cerevisiae mannoproteins form an external cell wall layer that determines wall porosity. J. Bacteriol. 159: 1018–1026.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • V. Ganeva
    • 1
  • B. Galutzov
    • 1
  • J. Teissié
    • 2
  1. 1.Faculty of BiologyUniversity of SofiaSofiaBulgaria
  2. 2.IPBS CNRS UMR5089Toulouse cédex 4France

Personalised recommendations