Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Immobilization of β-galactosidase retained in yeast: adhesion of the cells on a support

Summary

Treatment of Kluyveromyces cells with several polycations partially neutralized their negative charge, even after washing; chitosan at 0.5 mg/ml rendered the zeta potential positive. Adhesion of the cells to various supports (glass, polycarbonate, polystyrene) was promoted by treating the cells or the support with chitosan; this confirms the importance of electrostatic interactions in the adhesion process. When the support was treated, a dense and regular monolayer of adhering cells was obtained; when the cells were treated the adhering layer was more heterogeneous due to floc formation. Cell ghosts of K. lactis were prepared by CHCl3/ethanol permeabilization and glutaraldehyde treatment. Treating the ghosts with chitosan provided immobilization of 1.9 to 7.3×106 ghosts per cm2 according to the support; treating the support allowed to obtain 5.5 to 5.7×106 ghosts per cm2. The lactase activity of the immobilized ghosts was found to be comparable to that of ghosts in suspension, i.e. in the range of 0.9 to 2.2 U/mg dry cell weight, at 30°C, corresponding to 0.02 to 0.14 U/cm2 of support. A test in microreactor with a lactase solution showed that 50% of the initial activity was still found, after about 25 days of continuous operation at 30°C.

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

References

  1. Banerjee M, Chakravarty A, Majumdar K (1984) Immobilization of yeast cells containing β-Galactosidase. Biotechnol Bioeng 24:1839–1850

  2. Changui Ch, Doren A, Stone WEE, Mozes N, Rouxhet PG (1987) Surface properties of polycarbonate and promotion of yeast cell adhesion. J Chim Phys 84:275–281

  3. Decleire M, Van Huynh N, Motte J-C, De Cat W (1985) Hydrolysis of whey by whole cells of Kluyveromyces bulgaricus immobilized in calcium alginate gels and in hen egg white. Appl Microbiol Biotechnol 22:438–441

  4. Gekas V (1985) Hydrolysis of lactose: a literature review. Process Biochem Feb pp 2–11

  5. Hattori R, Hattori T (1985) Adsorptive phenomena involving bacterial cells and an anion exchange resin. J Gen Appl Microbiol 31:147–165

  6. Jaspers HTA, Christianse K, van Steveninck J (1975) An improved method for the preparation of yeast enzymes in situ. Biochem Biophys Res Comm 65:1434–1437

  7. Knorr D (1984) Use of chitinous polymers in food. Food Technol 1:85–97

  8. Knorr D, Teutonico RA (1986) Chitosan immobilization and permeabilization of Amaranthus tricolor cells. J Agr Food Chem 34:96–97

  9. Kokufuta E, Matsumoto W, Nakamura I (1982) Immobilization of Nitrosomonas europaea cells with polyelectrolyte complex. Biotechnol Bioeng 24:1591–1603

  10. Linko P, Poutanen K, Weckstrom L, Linko Y-Y (1980) Preparation and kinetic behavior of immobilized whole cell biocatalysts. Biochimie 62:387–394

  11. Rouxhet PG, Van Haecht JL, Didelez J, Gerard P, Briquet M (1981) Immobilization of yeast cells by entrapment and adhesion using siliceous materials. Enz Microbiol Technol 3:49–54

  12. Rouxhet PG (1985) New ways of enzyme utilization-immobilization of biocatalysts. Cerevisia 4:167–184

  13. Savage DC, Fletcher M (1985) Bacterial adhesion. Plenum Press, New York

  14. Stanley WL, Watters GG, Chan B, Mercer JM (1975) Lactase and other enzymes bound to chitin with glutaraldehyde. Biotechnol Bioeng 17:315–318

  15. Synowiecki J, Sikorski ZE, Naczk M (1981) The activity of immobilized enzymes on different krill chitin preparations. Biotechnol Bioeng 23:2211–2216

  16. Thonard Ph, Custinne M, Paquot M (1982) Zeta potential of yeast cell: application in cell immobilization. Enz Microbiol Technol 4:191–194

  17. Van Haecht JL, Bolipombo M, Rouxhet PG (1985) Immobilization of Saccharomyces cerevisiae by adhesion: treatment of the cells by Al ions. Biotechnol Bioeng 27:217–224

  18. Vlach D, Prenosil JE (1984) Yeast cells with high β-Galactosidase activity and their immobilization. J Molec Catal 26:173–185

  19. Vorlop KD, Klein J (1981) Formation of spherical chitosan biocatalysts by inotropic gelation. Biotechnol Lett 3:3–14

  20. Weetall HH (1985) Enzyme immobilized on inorganic supports. Trends Biotechnol 3:276–280

  21. Woychik JH, Wondolowski MV, Dahl KJ (1974) Purification and applications of immobilized β-Calactosidase of Saccharomyces lactis. In: Olson AC (ed) Immobilized enzymes in food and microbial processes, Plenum Press, New York

  22. Yoshiota T, Shimamura M (1986) Studies of polystyrene-based ion-exchange fiber. V. Immobilization of microorganisms by adsorption on a novel fiber-form anion-exchanger. Bull Chem Soc Japan 59:77–82

Download references

Author information

Correspondence to P. G. Rouxhet.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Champluvier, B., Kamp, B. & Rouxhet, P.G. Immobilization of β-galactosidase retained in yeast: adhesion of the cells on a support. Appl Microbiol Biotechnol 27, 464–469 (1988). https://doi.org/10.1007/BF00451614

Download citation

Keywords

  • Chitosan
  • Immobilization
  • Polystyrene
  • Ghost
  • Lactis