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KMID : 0545120070170040695
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 4 p.695 ~ p.700
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Characterization of Cyclofructans from Inulin by Saccharomyces cerevisiae Strain Displaying Cell-Surface Cycloinulooligosaccharide Fructanotransferase
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Kim Hyun-Jin
Lee Jae-Hyung
Kim Hyun-Chul
Lee Jin-Woo
Kim Yeon-Hee
Nam Soo-Wan
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Abstract
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The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus macerans (GenBank access code AF222787) was expressed on the cell surface of Saccharomyces cerevisiae by fusing with Aga2p linked to the membrane-anchored protein Aga1p. The surface display of CFTase was confirmed by immunofluorescence microscopy and enzymatic assay. The optimized reaction conditions of surface-displayed CFTase were as follows; pH, 8.0; temperature, 50oC; enzyme amount, 30 milliunit; substrate concentration, 5%; inulin source, Jerusalem artichoke. As a result of the reaction with inulin, cycloinulohexaose was produced as a major product along with cycloinuloheptilose and cycloinulooctaose as minor products.
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KEYWORD
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Cycloinulooligosaccharide fructanotransferase, Saccharomyces cerevisiae, cell surface display, cyclofructan, inulin
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