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Enhancement of beta-glucosidase activity on the cell-surface of sake yeast by disruption of SED1.
- Source :
-
Journal of bioscience and bioengineering [J Biosci Bioeng] 2010 May; Vol. 109 (5), pp. 442-6. Date of Electronic Publication: 2009 Dec 04. - Publication Year :
- 2010
-
Abstract
- We determined the genetic background that would result in a more optimal display of heterologously expressed beta-glucosidase (BGL) on the cell surface of yeast Saccharomyces cerevisiae. Amongst a collection of 28 strains carrying deletions in genes for glycosylphosphatidyl inositol (GPI)-anchored proteins, the Delta sed1 and Delta tos6 strains had significantly higher BGL-activity whilst maintaining wild type growth. Absence of Sed1p, which might facilitate incorporation of anchored BGL on the cell-surface, could also influence the activity of BGL on the cell surface with the heterologous gene being placed under the control of the SED1 promoter. For the evaluation of its industrial applicability we tested this system in heterologous and homogenous SED1-disruptants of sake yeast, a diploid S. cerevisiae strain, in which either the SED1 ORF or the complete gene including the promoter was deleted by use of the high-efficiency loss of heterozygosity method. Evaluation of disruptants displaying BGL showed that deletion of the SED1 ORF enhanced BGL activity on the cell surface, while additional deletion of the SED1 promoter increased further BGL activity on the cell surface. Compared to heterozygous disruption, homozygous disruption resulted generally in a higher BGL activity. Thus, homozygous deletion of both SED1 gene and promoter resulted in the most efficient display of BGL reaching a 1.6-fold increase of BGL-activity compared to wild type.<br /> ((c) 2009 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)
- Subjects :
- Enzyme Activation genetics
Gene Silencing physiology
Membrane Glycoproteins metabolism
Oryza microbiology
Saccharomyces cerevisiae classification
Saccharomyces cerevisiae Proteins metabolism
Species Specificity
beta-Glucosidase chemistry
Cell Membrane genetics
Genetic Enhancement methods
Membrane Glycoproteins genetics
Promoter Regions, Genetic genetics
Saccharomyces cerevisiae physiology
Saccharomyces cerevisiae Proteins genetics
beta-Glucosidase physiology
Subjects
Details
- Language :
- English
- ISSN :
- 1347-4421
- Volume :
- 109
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of bioscience and bioengineering
- Publication Type :
- Academic Journal
- Accession number :
- 20347765
- Full Text :
- https://doi.org/10.1016/j.jbiosc.2009.11.003