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J. Microbiol. Biotechnol. 2007; 17(4): 695-700

Published online April 28, 2007

Copyright © The Korean Society for Microbiology and Biotechnology.

Characterization of Cyclofructans from Inulin by Saccharomyces cerevisiae Strain Displaying Cell-Surface Cycloinulooligosaccharide Fructanotransferase

Kim, Hyun-Jin , Jae Hyung Lee , Hyun-Chul Kim 1, Jin-Woo Lee 2, Yeon-Hee Kim 3 and Soo-Wan Nam 4*

Department of Biomaterial Control (BK21 Program), Dong-Eui University, Busan 614-714, Korea, 1Bioneer Corp., Daejeon 306-220, Korea, 2Department of Biotechnology, Dong-A University, Busan 614-714, Korea, 3Department of Biotechnology, Osaka University, Osaka 565-0871, Japan, 4Department of Biomaterial Control (BK21 Program), Dong-Eui University, Busan 614-714, Korea Department of Biotechnology and Bioengineering, Dong-Eui University, Busan 614-714, Korea

Abstract

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, $50^{\circ}C$; 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 cycloinuloheptaose and cycloinulooctaose as minor products.

Keywords: Cycloinulooligosaccharide fructanotransferase, Saccharomyces cerevisiae, cell surface display, cyclofructan, inulin