The Optimization of Saccharomyces cerevisiae Expression System by Mutagenesis of Promoter and Host Strain

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An expression system with high efficiency is very important for recombinant proteins production in biopharmaceutical field. Saccharomyces cerevisiae is a food-graded eukayotic organism. The features of short generation time,simple culture condition and well-characterized manipulation techniques make yeast S. cerevisiae an attractive cell factory for production of heterologous protein. Here,for the purpose to improve the efficiency of p HR expression system which was constructed in our lab previously.,the promoter of p HR expression vector( PTEF) and host strain Y16 were modified by the way of error-prone PCR and mutagenesis respectively. After several rounds of screening,a mutant PTEFv1 with higher efficiency than the mother promoter PTEFwas obtained.Two modified yeast strains Y16-E14 and Y16-E19 were identified with higher productivity of heterologous protein than yeast Y16. Then PTEFv1 and yeast Y16-E14 were used to construct the novel p HR-N expression system. To evaluate the ability of p HR-N expression system,yeast green fluorescent protein( GFP) and human serum albumin( HSA) were chosed to be expressed intracellularly and extracellularly respectively. The results showed that p HR-N system had higher ability to produce either intracellular GFP or extracellular HSA than p HR system.The p HR-N yeast expression system provides a valuable resource for future application in recombinant protein production.