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KMID : 1234420090370040355
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Korean Journal of Microbiololgy and Biotechnology
2009 Volume.37 No. 4 p.355 ~ p.360
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Cell Surface Display of Arylsulfatase Gene from Pseudoalteromonas carageenovora in Saccharomyces cerevisiae
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Cho Eun-Soo
Kim Hyun-Jin
Jung So-A
Kim Jeong-Hwan
Kim Yeon-Hee
Nam Soo-Wan
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Abstract
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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 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-methylumbelliferyl-sulfate 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. The DNA ladder in agarose prepared from agar by this recombinant arylsulfatase showed similar resolution and migration patterns with a commercial agarose. This results revealed that arylsulfatase expressed on the cell surface of S. cerevisiae could be applicable to the economic production of electrophoretic-grade agarose.
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KEYWORD
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Cell surface display, Saccharomyces cerevisiae, arylsulfatase, Aga2p, agarose
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