title: Biochemical Characterization of Recombinant UDP-Glucose:Sterol 3-O-Glycosyltransferase from Micromonospora rhodorangea ATCC 31603 and Enzymatic Biosynthesis of Sterol-3-O-β-Glucosides.
abstract: A uridine diphosphate-glucose:sterol glycosyltransferase-encoding gene was isolated and cloned from the established fosmid library of Micromonospora rhodorangea ATCC 27932 that usually produces the aminoglycoside antibiotic geneticin. The gene consists of 1,185 base pairs and encodes a 41.4 kDa protein, which was heterologously expressed in Escherichia coli BL21(DE3). In silico analyses of the deduced gene product suggested that it is a member of the family 1 glycosyltransferases. The recombinant protein MrSGT was able to catalyze the transfer of a glucosyl moiety onto the C-3 hydroxy function in sterols (β-sitosterol, campesterol, and cholesterol), resulting in the corresponding steryl glucosides (β-sitosterol-3-O-β-D-glucoside, campesterol-3-O-β-D-glucoside, and cholesterol-3-O-β-D-glucoside). This enzyme prefers phytosterols to cholesterol, and also shows substrate flexibility to some extent, in that it could recognize a number of acceptor substrates.
authors.affiliation: Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University, Asan 31460, Republic of Korea.
authors.forename: Nguyen Huu
authors.initials: NH
authors.lastname: Hoang
authors.affiliation: School of Biosystem and Biomedical Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Sung-Yong
authors.initials: SY
authors.lastname: Hong
authors.affiliation: Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University, Asan 31460, Republic of Korea.
authors.forename: Nguyen Lan
authors.initials: NL
authors.lastname: Huong
authors.affiliation: School of Biosystem and Biomedical Science, Korea University, Seoul 02841, Republic of Korea.
authors.forename: Je Won
authors.initials: JW
authors.lastname: Park
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005960:Glucosides / Q000096:biosynthesis*; D016695:Glycosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008847:Micromonospora / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013261:Sterols / Q000378:metabolism; D014532:Uridine Diphosphate Glucose / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005960:Glucosides; D011994:Recombinant Proteins; D013261:Sterols; D016695:Glycosyltransferases; D014532:Uridine Diphosphate Glucose
keywords: Micromonospora rhodorangea; UDP-glucose sterol glycosyltransferase; phytosterol-3-O-β-D-glucosides