TY - JOUR
T1 - Biochemical characterization of recombinant UDP-glucose
T2 - Sterol 3-O-Glycosyltransferase from Micromonospora rhodorangea ATCC 31603 and enzymatic biosynthesis of sterol-3-O-β-glucosides
AU - Hoang, Nguyen Huu
AU - Hong, Sung Yong
AU - Huong, Nguyen Lan
AU - Park, Je Won
N1 - Funding Information:
This work was supported by the National Research Foundation of Korea grant (2015R1A2A2A01002524) funded by the Ministry of Science, ICT and Future Planning, and by the grant (PJ011066) funded by the Next-Generation BioGreen21 program, Rural Development Administration.
Publisher Copyright:
© 2016 by The Korean Society for Microbiology and Biotechnology.
PY - 2015/12/8
Y1 - 2015/12/8
N2 - 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.
AB - 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.
KW - Micromonospora rhodorangea
KW - Phytosterol-3-O-β-D-glucosides
KW - UDP-glucose sterol glycosyltransferase
UR - http://www.scopus.com/inward/record.url?scp=84961659749&partnerID=8YFLogxK
U2 - 10.4014/jmb.1511.11003
DO - 10.4014/jmb.1511.11003
M3 - Article
C2 - 26643965
AN - SCOPUS:84961659749
SN - 1017-7825
VL - 26
SP - 477
EP - 482
JO - Journal of Microbiology and Biotechnology
JF - Journal of Microbiology and Biotechnology
IS - 3
ER -