title: Identification and functional analysis of 2-hydroxyflavanone C-glucosyltransferase in soybean (Glycine max).
abstract: C-Glucosyltransferase is an enzyme that mediates carbon-carbon bond formation to generate C-glucoside metabolites. Although it has been identified in several plant species, the catalytic amino acid residues required for C-glucosylation activity remain obscure. Here, we identified a 2-hydroxyflavanone C-glucosyltransferase (UGT708D1) in soybean. We found that three residues, His20, Asp85, and Arg292, of UGT708D1 were located at the predicted active site and evolutionarily conserved. The substitution of Asp85 or Arg292 with alanine destroyed C-glucosyltransferase activity, whereas the substitution of His20 with alanine abolished C-glucosyltransferase activity but enabled O-glucosyltransferase activity. The catalytic mechanism is discussed on the basis of the findings.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamada-oka, Suita City, Osaka 565-0871, Japan. Electronic address: hiradeyoshihiro@gmail.com.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Hirade
authors.affiliation: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita City, Osaka 565-0871, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Kotoku
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya City, Aichi 467-8603, Japan.
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Terasaka
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamada-oka, Suita City, Osaka 565-0871, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Saijo-Hamano
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamada-oka, Suita City, Osaka 565-0871, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Fukumoto
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya City, Aichi 467-8603, Japan; The Kochi Prefectural Makino Botanical Garden, 4200-6 Godaisan, Kochi City, Kochi 781-8125, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Mizukami
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis; D020134:Catalytic Domain; D002851:Chromatography, High Pressure Liquid; D005964:Glucosyltransferases / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D005964:Glucosyltransferases
keywords: 2-Hydroxyflavanone C-glucosyltransferase; C-Glucoside biosynthesis; Catalytic residues; Enzymatic aryl-C-glucosylation; Secondary metabolites; Structural modeling