Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research
  • Published:

Selective Release of Recombinant Protein Particles (VLPs) from Yeast Using a Pure Lytic Glucanase Enzyme

Bio/Technology volume 11, pages 214–217 (1993)Cite this article

Abstract

We have used a pure lytic glucanase enzyme to selectively release recombinant 60 nm protein particles (virus like particles or VLPs) from yeast cells. Although the protease components of the lytic enzyme complexes were found to degrade the VLPs, purified glucanase enzymes from these complexes (derived from Cytophaga sp. and Oerskovia sp.) produced cell lysis without degradation and released the VLPs in the absence of mercaptoethanol, a reducing agent commonly used in cell lysis. The Oerskovia glucanase enzyme released the recombinant protein particles selectively as it only produced ca. 17% cell lysis compared to the use of the total lytic enzyme preparation. The use of osmotic supports did not improve the recovery of VLPs, however, treatment of the enzymatically lysed pellet with Triton X-100 did increase the amount of VLPs released. This ‘selectivity’, which results in the release of the recombinant particles with only a fraction of contaminating proteins, represents an improvement over presently used mechanical or enzymatic cell disruption processes.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Andrews, B.A., Huang, R.-B. and Asenjo, J.A. 1990. Differential product release from yeast cells by selective enzymatic lysis. 21–28. In: Separations for Biotechnology 2.Pyle, D. L. (Ed.). Elsevier Applied Science, Cambridge, UK.

    Chapter  Google Scholar 

  2. Huang, R.-B., Andrews, B.A. and Asenjo, J.A. 1991. Differentia] product release from yeast: A new technique for selective product recovery from microbial cells. Biotechnol. Bioeng. 38: 977–985.

    Article  CAS  Google Scholar 

  3. Cameron, J.R., Loh, E.Y. and Davis, R.W. 1979. Evidence for transposition of dispersed repetitive DNA families in yeast. Cell 16: 739–751.

    Article  CAS  Google Scholar 

  4. Garfinkel, D.J., Boeke, J.D. and Fink, G.R. 1985. Ty element transposition: Reverse transcriptase and virus-like particles. Cell 42: 507–517.

    Article  CAS  Google Scholar 

  5. Kingsman, S.M. and Kingsman, A.J. 1988. Polyvalent recombinant antigens: A new vaccine strategy. Vaccine 7: 304–306.

    Article  Google Scholar 

  6. Boeke, J.D., Garfinkel, D.J., Styles, C.A. and Fink, G.R. 1985. Ty elements transpose through an RNA intermediate. Cell 70: 491–500.

    Article  Google Scholar 

  7. Adams, S.E., Kingsman, S.M. and Kingsman, A.J. 1987. The yeast Ty element: Recent advances in the study of a model retro-element. Bioessays 7: 3–9.

    Article  CAS  Google Scholar 

  8. Asenjo, J.A., Andrews, B.A. and Pitts, J.M. 1988. Design of enzyme systems for selective product release from microbial cells; isolation of a recombinant protein from yeast. Ann. N.Y. Acad. Sci. 52: 140–152.

    Article  Google Scholar 

  9. Huang, R.-B. 1990. Selective separation of yeast proteins by differential product release, enzymatic lysis and aqueous two-phase systems. Ph.D. Thesis, University of Reading, UK.

  10. Ventom, A.M. and Asenjo, J.A. 1990. Purification of the major glucanase of Oerskovia xanthineolytica LL-GI09. Biotechnol. Techniques 4: 165–170.

    CAS  Google Scholar 

  11. Ventom, A.M. and Asenjo, J.A. 1990. Two extracellular proteases from Oerskovia xanthineolyticaLL-G109. Biotechnol. Techniques 4: 171–176.

    CAS  Google Scholar 

  12. Ventom, A.M. and Asenjo, J.A. 1991. Characterization of yeast lytic enzymes from Oerskovia xanthineolyticaLL-G109. Enzyme Microb. Technol. 13: 71–75.

    Article  CAS  Google Scholar 

  13. Andrews, B.A. and Asenjo, J.A. 1987. Enzymatic lysis and disruption of microbial cells. Trends in Biotechnol. 5: 273–277.

    Article  CAS  Google Scholar 

  14. Varela, O. 1991. Maximization of enzyme (glucanase) production using optimized fed-batch cultures. M.Sc. Dissertation, University of Reading-Universidad Autonoma de Barcelona (ERASMUS programme).

  15. Shen, S.H., Chretien, P., Bastien, L. and Slilaty, S.N. 1991. Primary sequence of the glucanase gene from Oerskovia xanthineolytica. J. Biol. Chem. 266: 1058–1063.

    CAS  PubMed  Google Scholar 

  16. Andrews, B.A. and Asenjo, J.A. 1986. Synthesis and regulation of extracellular β(1–3) glucanase and protease by Cytophaga sp. in batch and continuous culture. Biotechnol. Bioeng. 28: 1366–1375.

    Article  CAS  Google Scholar 

  17. Andrews, B.A. and Asenjo, J.A. 1987. Continuous culture studies of synthesis and regulation of extracellular β(1–3) glucanase and protease enzymes from Oerskovia xanthineolytica. Biotechnol. Bioeng. 30: 628–637.

    Article  CAS  Google Scholar 

  18. Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354.

    Article  CAS  Google Scholar 

  19. Hawkes, R. 1987. The dot immunobinding assay. Methods in Enzymol. 121: 484–491.

    Article  Google Scholar 

Download references

Author information

Author notes

  1. A. M. Ventom

    Present address: Oxford Glycosystems, Oxford, England

  2. R. -B. Huang

    Present address: The Research Centre, Guanxi Agricultural College, Nanning, Guanxi, 530005, People's Republic of China

  3. J. A. Asenjo and B. A. Andrews: Corresponding authors.

Authors and Affiliations

  1. Biochemical Engineering Laboratory, University of Reading, P.O. Box 226, Reading, RG6 2UQ, England

    J. A. Asenjo & B. A. Andrews

Authors
  1. J. A. Asenjo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. M. Ventom
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. -B. Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B. A. Andrews
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Asenjo, J., Ventom, A., Huang, RB. et al. Selective Release of Recombinant Protein Particles (VLPs) from Yeast Using a Pure Lytic Glucanase Enzyme. Nat Biotechnol 11, 214–217 (1993). https://doi.org/10.1038/nbt0293-214

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0293-214

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing