Characterization of cyclofructans from inulin by Saccharomyces cerevisiae Strain displaying cell-surface cycloinulooligosaccharide fructanotransferase.
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 degrees 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.
Kim
Hyun-Jin
HJ
Department of Biomaterial Control (BK21 Program), Dong-Eui University, Busan 614-714, Korea.
Lee
Jae Hyung
JH
Kim
Hyun-Chul
HC
Lee
Jin-Woo
JW
Kim
Yeon-Hee
YH
Nam
Soo-Wan
SW
D005630:Fructans / Q000378:metabolism*; D006602:Hexosyltransferases / Q000502:physiology*; D007150:Immunohistochemistry; D007444:Inulin / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D005630:Fructans; D007444:Inulin; D006602:Hexosyltransferases; C062589:cycloinulo-oligosaccharide fructanotransferase