Enzymatic total synthesis of gibberellin A4 from acetate

Yoshinori Sugai, Sho Miyazaki, Shinichiro Mukai, Isamu Yumoto, Masahiro Natsume, Hiroshi Kawaide

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Natural terpenoids have elaborate structures and various bioactivities, making difficult their synthesis and labeling with isotopes. We report here the enzymatic total synthesis of plant hormone gibberellins (GAs) with recombinant biosynthetic enzymes from stable isotopelabeled acetate. Mevalonate (MVA) is a key intermediate for the terpenoid biosynthetic pathway. 13C-MVA was synthesized from 13C-acetate via acetyl-CoA, using four enzymes or fermentation with a MVA-secreted yeast. The diterpene hydrocarbon, ent-kaurene, was synthesized from 13C-acetate and 13C-MVA with ten and six recombinant enzymes in one test tube, respectively. Four recombinant enzymes, P450 monooxygenases and soluble dioxygenases involved in the GA 4 biosynthesis from ent-kaurene via GA12were prepared in yeast and Escherichia coli. All intermediates and the final product GA 4 were uniformly labeled with 13C without dilution by natural abundance when [U-13C2] acetate was used. The 13C-NMR and MS data for [U-13C20] ent-kaurene confirmed 13C-13C coupling, and no dilution with the 12C atom was observed.

Original languageEnglish
Pages (from-to)128-135
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume75
Issue number1
DOIs
Publication statusPublished - 2011 Jan 28
Externally publishedYes

Fingerprint

Mevalonic Acid
Kaurane Diterpenes
Acetates
Terpenes
Yeast
Dilution
Enzymes
Yeasts
Isotope Labeling
Gibberellins
Dioxygenases
Acetyl Coenzyme A
Plant Growth Regulators
Diterpenes
Biosynthesis
Biosynthetic Pathways
Hydrocarbons
Mixed Function Oxygenases
Bioactivity
Isotopes

Keywords

  • C-NMR
  • Biosynthesis
  • Isotope
  • Recombinant enzyme
  • Terpenoid

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Enzymatic total synthesis of gibberellin A4 from acetate. / Sugai, Yoshinori; Miyazaki, Sho; Mukai, Shinichiro; Yumoto, Isamu; Natsume, Masahiro; Kawaide, Hiroshi.

In: Bioscience, Biotechnology and Biochemistry, Vol. 75, No. 1, 28.01.2011, p. 128-135.

Research output: Contribution to journalArticle

Sugai, Yoshinori ; Miyazaki, Sho ; Mukai, Shinichiro ; Yumoto, Isamu ; Natsume, Masahiro ; Kawaide, Hiroshi. / Enzymatic total synthesis of gibberellin A4 from acetate. In: Bioscience, Biotechnology and Biochemistry. 2011 ; Vol. 75, No. 1. pp. 128-135.
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