This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sagt, C. M. J. Articles by Verrips, C. T. Search for Related Content PubMed PubMed Citation Articles by Sagt, C. M. J. Articles by Verrips, C. T. Agricola Articles by Sagt, C. M. J. Articles by Verrips, C. T.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2000, p. 4940-4944, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Introduction of an N-Glycosylation Site Increases Secretion of Heterologous Proteins in Yeasts

C. M. J. Sagt,¹ B. Kleizen,¹ R. Verwaal,¹ M. D. M. de Jong,¹ W. H. Müller,¹ A. Smits,² C. Visser,² J. Boonstra,^1,* A. J. Verkleij,¹ and C. T. Verrips^1,2

Department of Molecular Cell Biology and the Institute of Biomembranes, Utrecht University, 3584 CH Utrecht,¹ and Unilever Research Laboratory Vlaardingen, 3133 AT Vlaardingen,² The Netherlands

Received 21 March 2000/Accepted 28 June 2000

Saccharomyces cerevisiae is often used to produce heterologous proteins that are preferentially secreted to increase economic feasibility. We used N-glycosylation as a tool to enhance protein secretion. Secretion of cutinase, a lipase, and llama V_HH antibody fragments by S. cerevisiae or Pichia pastoris improved following the introduction of an N-glycosylation site. When we introduced an N-glycosylation consensus sequence in the N-terminal region of a hydrophobic cutinase, secretion increased fivefold. If an N-glycosylation site was introduced in the C-terminal region, however, secretion increased only 1.8-fold. These results indicate that the use of N glycosylation can significantly enhance heterologous protein secretion.

---

^* Corresponding author. Mailing address: Department of Molecular Cell Biology and the Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. Phone: 31 30 2533189. Fax: 31 30 2513655. E-mail: J.Boonstra{at}bio.uu.nl.

---

Applied and Environmental Microbiology, November 2000, p. 4940-4944, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

• Levasseur, A., Navarro, D., Punt, P. J., Belaich, J.-P., Asther, M., Record, E. (2005). Construction of Engineered Bifunctional Enzymes and Their Overproduction in Aspergillus niger for Improved Enzymatic Tools To Degrade Agricultural By-Products. Appl. Environ. Microbiol. 71: 8132-8140 [Abstract] [Full Text]  
• Masaki, K., Kamini, N. R., Ikeda, H., Iefuji, H. (2005). Cutinase-Like Enzyme from the Yeast Cryptococcus sp. Strain S-2 Hydrolyzes Polylactic Acid and Other Biodegradable Plastics. Appl. Environ. Microbiol. 71: 7548-7550 [Abstract] [Full Text]  
• Valkonen, M., Penttila, M., Saloheimo, M. (2003). Effects of Inactivation and Constitutive Expression of the Unfolded- Protein Response Pathway on Protein Production in the Yeast Saccharomyces cerevisiae. Appl. Environ. Microbiol. 69: 2065-2072 [Abstract] [Full Text]  
• Sagt, C. M. J., Muller, W. H., van der Heide, L., Boonstra, J., Verkleij, A. J., Verrips, C. T. (2002). Impaired Cutinase Secretion in Saccharomyces cerevisiae Induces Irregular Endoplasmic Reticulum (ER) Membrane Proliferation, Oxidative Stress, and ER-Associated Degradation. Appl. Environ. Microbiol. 68: 2155-2160 [Abstract] [Full Text]