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 Donnini, C. Articles by Palleschi, C. Search for Related Content PubMed PubMed Citation Articles by Donnini, C. Articles by Palleschi, C. Agricola Articles by Donnini, C. Articles by Palleschi, C.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, May 2004, p. 2632-2638, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2632-2638.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Improved Production of Heterologous Proteins by a Glucose Repression-Defective Mutant of Kluyveromyces lactis

Department of Genetics Anthropology Evolution, University of Parma, Parma,¹ Department of Developmental and Cell Biology, University of Rome La Sapienza, Rome,² Department of General Physiology and Biochemistry, University of Milan, Milan, Italy³

Received 31 October 2003/ Accepted 27 January 2004

The secreted production of heterologous proteins in Kluyveromyces lactis was studied. A glucoamylase (GAA) from the yeast Arxula adeninivorans was used as a reporter protein for the study of the secretion efficiencies of several wild-type and mutant strains of K. lactis. The expression of the reporter protein was placed under the control of the strong promoter of the glyceraldehyde-3-phosphate dehydrogenase of Saccharomyces cerevisiae. Among the laboratory strains tested, strain JA6 was the best producer of GAA. Since this strain is known to be highly sensitive to glucose repression and since this is an undesired trait for biomass-oriented applications, we examined heterologous protein production by using glucose repression-defective mutants isolated from this strain. One of them, a mutant carrying a dgr151-1 mutation, showed a significantly improved capability of producing heterologous proteins such as GAA, human serum albumin, and human interleukin-1ß compared to the parent strain. dgr151-1 is an allele of RAG5, the gene encoding the only hexokinase present in K. lactis (a homologue of S. cerevisiae HXK2). The mutation in this strain was mapped to nucleotide position +527, resulting in a change from glycine to aspartic acid within the highly conserved kinase domain. Cells carrying the dgr151-1 allele also showed a reduction in N- and O-glycosylation. Therefore, the dgr151 strain may be a promising host for the production of heterologous proteins, especially when the hyperglycosylation of recombinant proteins must be avoided.

This article has been cited by other articles:

• Lodi, T., Neglia, B., Donnini, C. (2005). Secretion of Human Serum Albumin by Kluyveromyces lactis Overexpressing KlPDI1 and KlERO1. Appl. Environ. Microbiol. 71: 4359-4363 [Abstract] [Full Text]