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

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2005, p. 4359-4363, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4359-4363.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Secretion of Human Serum Albumin by Kluyveromyces lactis Overexpressing KlPDI1 and KlERO1

Tiziana Lodi, Barbara Neglia, and Claudia Donnini*

Department of Genetics, Anthropology, and Evolution, University of Parma, Parma, Italy

Received 20 October 2004/ Accepted 4 March 2005

The control of protein conformation during translocation through the endoplasmic reticulum is often a bottleneck for heterologous protein production. The core pathway of the oxidative folding machinery includes two conserved proteins: Pdi1p and Ero1p. We increased the dosage of the genes encoding these proteins in the yeast Kluyveromyces lactis and evaluated the secretion of heterologous proteins. KlERO1, an orthologue of Saccharomyces cerevisiae ERO1, was cloned by functional complementation of the ts phenotype of an Scero1 mutant. The expression of KlERO1 was induced by treatment of the cells with dithiothreitol and by overexpression of human serum albumin (HSA), a disulfide bond-rich protein. Duplication of either PDI1 or ERO1 led to a similar increase in HSA yield. Duplication of both genes accelerated the secretion of HSA and improved cell growth rate and yield. Increasing the dosage of KlERO1 did not affect the production of human interleukin 1ß, a protein that has no disulfide bridges. The results confirm that the ERO1 genes of S. cerevisiae and K. lactis are functionally similar even though portions of their coding sequence are quite different and the phenotypes of mutants overexpressing the genes differ. The marked effects of KlERO1 copy number on the expression of heterologous proteins with a high number of disulfide bridges suggests that control of KlERO1 and KlPDI1 is important for the production of high levels of heterologous proteins of this type.

* Corresponding author. Mailing address: Department of Genetics, Anthropology, and Evolution, University of Parma, Parco Area delle Scienze 11/A, I-43100, Parma, Italy. Phone: 39 0521 905602. Fax: 39 0521 905604. E-mail: claudia.donnini{at}unipr.it.

Applied and Environmental Microbiology, August 2005, p. 4359-4363, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4359-4363.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Raimondi, S., Zanni, E., Talora, C., Rossi, M., Palleschi, C., Uccelletti, D. (2008). SOD1, a New Kluyveromyces lactis Helper Gene for Heterologous Protein Secretion. Appl. Environ. Microbiol. 74: 7130-7137 [Abstract] [Full Text]  
  • Gasser, B., Sauer, M., Maurer, M., Stadlmayr, G., Mattanovich, D. (2007). Transcriptomics-Based Identification of Novel Factors Enhancing Heterologous Protein Secretion in Yeasts. Appl. Environ. Microbiol. 73: 6499-6507 [Abstract] [Full Text]  
  • Read, J. D., Colussi, P. A., Ganatra, M. B., Taron, C. H. (2007). Acetamide Selection of Kluyveromyces lactis Cells Transformed with an Integrative Vector Leads to High-Frequency Formation of Multicopy Strains. Appl. Environ. Microbiol. 73: 5088-5096 [Abstract] [Full Text]  
  • Wentz, A. E., Shusta, E. V. (2007). A Novel High-Throughput Screen Reveals Yeast Genes That Increase Secretion of Heterologous Proteins. Appl. Environ. Microbiol. 73: 1189-1198 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»