Project description:Saccharomyces cerevisiae has been used as a secretion host for production of various products, including pharmaceuticals. However, few antibody molecules have been functionally expressed in S. cerevisiae due to the incompatible surface glycosylation. Our laboratory previously isolated a group of yeast mutant strains with different α-amylase secretory capacities, and these evolved strains have showed advantages for production of some heterologous proteins. However, it is not known whether these secretory strains are generally suitable for pharmaceutical protein production. Here, three non-glycosylated antibody fragments with different configurations (Ran-Fab fragment Ranibizumab, Pex-the scFv peptide Pexelizumab, and Nan-a single V-type domain) were successfully expressed and secreted in three background strains with different secretory capacities, including HA (wild type), MA (evolved strain), and LA (evolved strain). However, the secretion of Ran and Nan were positively correlated with the strains’ secretory capacity, while Pex was most efficiently secreted in the parental strain. Therefore, transcriptional analysis was performed to explore the fundamental changes triggered by the expression of the different pharmaceutical proteins in these selected yeast strains.
Project description:We have employed whole genome microarray expression profiling as a discovery platform to identify genes implicated in the resistance to cobalt in Saccharomyces cerevisiae. The evolved strains and the wild type were harvested in exponential phase
Project description:In this study, we have developed a highly SO2-stress-resistant yeast (Saccharomyces cerevisiae) strain [F3] using evolutionary engineering, by successive batch selection at gradually increased SO2 levels. The evolved F3 strain was resistant to 1.0 mM SO2 stress, which was strongly inhibitory to the reference strain. Whole-transcriptomic analysis of F3 was performed with respect to its reference strain to determine differences in gene expression levels between the two strains. Saccharomyces cerevisiae
Project description:We have employed whole genome microarray expression profiling as a discovery platform to identify genes implicated in the resistance to cobalt in Saccharomyces cerevisiae. The evolved strains and the wild type were harvested in exponential phase WT (CEN.PK), the evolved strain (CI25E) and the evolved strain delta Cot1 harvested in exponential phase (DO 600nm=3) ; three independant replicate for each.
