Expression of the human blood coagulation protein Factor XIIIa in Saccharomyces cerevisiae: dependence of the expression levels from host-vector systems and medium conditions

• Published in: Applied microbiology and biotechnology Vol. 34; no. 6; pp. 756 - 764
• Main Authors: Broker, M, Bauml, O, Gotting, A, Ochs, J, Bodenbenner, M, Amann, E
• Format: Journal Article
• Language: English
• Published: Germany 01.03.1991
• Subjects: blood coagulation factors; Chromosomes, Fungal - physiology; Cloning, Molecular; Culture Media; DNA, Fungal - chemistry; expression vectors; fermentation; galactose; Galactose - genetics; Galactosidases - biosynthesis; Galactosidases - genetics; Gene Expression; genetic regulation; genetic transformation; glucose; humans; Milk Proteins - genetics; Plasmids; promoter regions; Promoter Regions, Genetic; Recombination, Genetic; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; structural genes; transcription (genetics); Transglutaminases - biosynthesis; Transglutaminases - genetics; vectors; Whey Proteins
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/BF00169346

The human blood coagulation protein Factor XIIIa (FXIIIa) was expressed in Saccharomyces cerevisiae employing Escherichia coli-yeast shuttle vectors based on a 2-micro plasmid. Several factors affecting high production yield of recombinant FXIIIa were analysed. The use of the regulatable GAL-CYC1 hybrid promoter resulted in higher FXIIIa expression when compared with the constitutive ADCI promoter. Screening for suitable yeast strains for expression of FXIIIa under the transcriptional control of the GAL-CYC1 hybrid promoter revealed a broad spectrum of productivity. No obvious correlation between the expression rate and the genetic markers of the strains could be identified. The medium composition markedly influenced the FXIIIa expression rates. The expression of FXIIIa was strictly regulated by the carbon source. Glucose as the only sugar and energy source repressed the synthesis of FXIIIa, whereas addition of galactose induced FXIIIa expression. Special feeding schemes resulted in a productivity of up to 100 mg FXIIIa/1 in shake flasks.