title: Acceptor Specificity of Amylosucrase from Deinococcus radiopugnans and Its Application for Synthesis of Rutin Derivatives.
abstract: The transglycosylation activity of amylosucrase (ASase) has received significant attention owing to its use of an inexpensive donor, sucrose, and broad acceptor specificity, including glycone and aglycone compounds. The transglycosylation reaction of recombinant ASase fromDeinococcus radiopugnans(DRpAS) was investigated using various phenolic compounds, and quercetin-3-O-rutinoside (rutin) was found to be the most suitable acceptor molecule used by DRpAS. Two amino acid residues in DRpAS variants (DRpAS Q299K and DRpAS Q299R), assumed to be involved in acceptor binding, were constructed by site-directed mutagenesis. Intriguingly, DRpAS Q299K and DRpAS Q299R produced 10-fold and 4-fold higher levels of rutin transglycosylation product than did the wild-type (WT) DRpAS, respectively. According to in silico molecular docking analysis, the lysine residue at position 299 in the mutants enables rutin to more easily position inside the active pocket of the mutant enzyme than in that of the WT, due to conformational changes in loop 4.
authors.affiliation: Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 17104, Republic of Korea.|Korea Food Research Institute, Seongnam 13539, Republic of Korea.
authors.forename: Myo-Deok
authors.initials: MD
authors.lastname: Kim
authors.affiliation: Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 17104, Republic of Korea.
authors.forename: Dong-Hyun
authors.initials: DH
authors.lastname: Jung
authors.affiliation: Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 17104, Republic of Korea.|Korea Food Research Institute, Seongnam 13539, Republic of Korea.
authors.forename: Dong-Ho
authors.initials: DH
authors.lastname: Seo
authors.affiliation: Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 17104, Republic of Korea.|Research Division for Biotechnology, Korea Atomic Energy Research Institute, Jeongeup 56212, Republic of Korea.
authors.forename: Jong-Hyun
authors.initials: JH
authors.lastname: Jung
authors.affiliation: Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Mainz 55128, Germany.
authors.forename: Ean-Jeong
authors.initials: EJ
authors.lastname: Seo
authors.affiliation: Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 17104, Republic of Korea.
authors.forename: Nam-In
authors.initials: NI
authors.lastname: Baek
authors.affiliation: Department of Food Science and Technology, Sejong University, Seoul 05006, Republic of Korea.
authors.forename: Sang-Ho
authors.initials: SH
authors.lastname: Yoo
authors.affiliation: Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 17104, Republic of Korea.
authors.forename: Cheon-Seok
authors.initials: CS
authors.lastname: Park
mesh_terms: D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D034301:Deinococcus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D007700:Kinetics; D062105:Molecular Docking Simulation; D012431:Rutin / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D012431:Rutin; D005964:Glucosyltransferases; C019700:amylosucrase
keywords: Acceptor specificity; Deinococcus radiopugnans; In silico molecular docking; amylosucrase (ASase); quercetin-3-O-rutinoside (rutin); transglycosylation