Production of a new sucrose derivative by transglycosylation of recombinant Sulfolobus shibatae beta-glycosidase.
The gene encoding beta-glycosidase of the hyperthermophilic archaea Sulfolobus shibatae (SSG) was expressed in Escherichia coli. Recombinant SSG (referred to as rSSG hereafter) was efficiently purified, and its transglycosylation activity was tested with lactose as a donor and various sugars as acceptors. When sucrose was used as an acceptor, we found a distinct intermolecular transglycosylation product and confirmed its presence by TLC and high performance anion exchange chromatography (HPAEC). The sucrose transglycosylation product was isolated by paper chromatography, and its chemical structure was determined by 1H and 13C NMR. The sucrose transfer product was determined to be beta-D-galactopyranosyl-(1-->6)-alpha-D-glucopyranosyl-beta-d-fructofuranoside with a galactose molecule linked to sucrose via a beta-(1-->6)-glycosidic bond.
Park
Na-Young
NY
Department of Food Science and Biotechnology and Institute of Life Sciences and Resources, KyungHee University, Yongin 449-701, Republic of Korea.
Baek
Nam-In
NI
Cha
Jaeho
J
Lee
Soo-Bok
SB
Auh
Joong-Hyuck
JH
Park
Cheon-Seok
CS
D002855:Chromatography, Thin Layer; D005959:Glucosidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006031:Glycosylation; D009682:Magnetic Resonance Spectroscopy; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013395:Sucrose / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D048229:Sulfolobus solfataricus / Q000201:enzymology* / Q000235:genetics
D011994:Recombinant Proteins; D013395:Sucrose; D005959:Glucosidases; C104851:Sulfolobus solfataricus beta-glycosidase