earticle

영어

Cell surface display engineering is expected to be useful for the segregation of recombinant polypeptides and for the construction of microbial biocatalysts. Expression of proteins on the cell surface of Saccharomyces cerevisiae offers more advantages than other microbial systems, since it allows the folding and glycosylation of expressed heterologous eukaryotic proteins and can be subjected to many genetic manipulations. In this study, the arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was expressed on the cell surface of S. cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The arylsulfatase gene was subcloned into the surface display vector, pCTcon (GAL1 promoter). The constructed plasmid, pCTAST (7.1 kb), was introduced to S. cerevisiae EBY100 cell, and yeast transformants on YPDG plate showed the hydrolyzing activity for 4-methylumbelliferylsulfate
and p-nitrophenyl-sulfate. When S. cerevisiae EBY100/pCTAST was grown on YPDG medium, the arylsulfatase activity of cell pellet reached about 1.2 unit/ml, whereas no extracellular arylsulfatase activity was detected.