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Functional specialization of UDP-glycosyltransferase 73P12 in licorice to produce a sweet triterpenoid saponin, glycyrrhizin.
Nomura, Yuhta; Seki, Hikaru; Suzuki, Tomonori; Ohyama, Kiyoshi; Mizutani, Masaharu; Kaku, Tomomi; Tamura, Keita; Ono, Eiichiro; Horikawa, Manabu; Sudo, Hiroshi; Hayashi, Hiroaki; Saito, Kazuki; Muranaka, Toshiya.
Afiliação
  • Nomura Y; Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
  • Seki H; Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
  • Suzuki T; RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.
  • Ohyama K; Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
  • Mizutani M; RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.
  • Kaku T; Department of Chemistry and Materials Science, Tokyo Institute of Technology, 2-12-1 O-okayama, Meguro-ku, Tokyo, 152-8551, Japan.
  • Tamura K; Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo, 657-8501, Japan.
  • Ono E; Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
  • Horikawa M; Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
  • Sudo H; Suntory Global Innovation Center Ltd, Research Institute, 8-1-1 Seikadai, Seika-cho, Soraku-gun, Kyoto, 619-0284, Japan.
  • Hayashi H; Suntory Foundation for Life Sciences, Bioorganic Research Institute, 8-1-1 Seikadai, Seika-cho, Soraku-gun, Kyoto, 619-0284, Japan.
  • Saito K; Tokiwa Phytochemical Co., Ltd, 158 Kinoko, Sakura, Chiba, 285-0801, Japan.
  • Muranaka T; Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8675, Japan.
Plant J ; 99(6): 1127-1143, 2019 09.
Article em En | MEDLINE | ID: mdl-31095780
ABSTRACT
Glycyrrhizin, a sweet triterpenoid saponin found in the roots and stolons of Glycyrrhiza species (licorice), is an important active ingredient in traditional herbal medicine. We previously identified two cytochrome P450 monooxygenases, CYP88D6 and CYP72A154, that produce an aglycone of glycyrrhizin, glycyrrhetinic acid, in Glycyrrhiza uralensis. The sugar moiety of glycyrrhizin, which is composed of two glucuronic acids, makes it sweet and reduces its side-effects. Here, we report that UDP-glycosyltransferase (UGT) 73P12 catalyzes the second glucuronosylation as the final step of glycyrrhizin biosynthesis in G. uralensis; the UGT73P12 produced glycyrrhizin by transferring a glucuronosyl moiety of UDP-glucuronic acid to glycyrrhetinic acid 3-O-monoglucuronide. We also obtained a natural variant of UGT73P12 from a glycyrrhizin-deficient (83-555) strain of G. uralensis. The natural variant showed loss of specificity for UDP-glucuronic acid and resulted in the production of an alternative saponin, glucoglycyrrhizin. These results are consistent with the chemical phenotype of the 83-555 strain, and suggest the contribution of UGT73P12 to glycyrrhizin biosynthesis in planta. Furthermore, we identified Arg32 as the essential residue of UGT73P12 that provides high specificity for UDP-glucuronic acid. These results strongly suggest the existence of an electrostatic interaction between the positively charged Arg32 and the negatively charged carboxy group of UDP-glucuronic acid. The functional arginine residue and resultant specificity for UDP-glucuronic acid are unique to UGT73P12 in the UGT73P subfamily. Our findings demonstrate the functional specialization of UGT73P12 for glycyrrhizin biosynthesis during divergent evolution, and provide mechanistic insights into UDP-sugar selectivity for the rational engineering of sweet triterpenoid saponins.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Glicosiltransferases / Ácido Glicirrízico / Glycyrrhiza uralensis Tipo de estudo: Prognostic_studies Idioma: En Revista: Plant J Assunto da revista: BIOLOGIA MOLECULAR / BOTANICA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Japão

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Glicosiltransferases / Ácido Glicirrízico / Glycyrrhiza uralensis Tipo de estudo: Prognostic_studies Idioma: En Revista: Plant J Assunto da revista: BIOLOGIA MOLECULAR / BOTANICA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Japão