In vitro assay and inhibition of 9-cis-epoxycarotenoid dioxygenase (NCED) from Solanum lycopersicum and Zea mays

Methods Enzymol. 2024:704:291-312. doi: 10.1016/bs.mie.2024.05.012. Epub 2024 Jun 6.

Abstract

The article reports methods for the expression and assay of 9-cis-epoxycarotenoid cleavage dioxygenase (NCED), an enzyme involved in the biosynthesis of phytohormone abscisic acid in plants. A method for the preparation of the unstable substrate 9'-cis-neoxanthin from fresh spinach is described. The inhibition of Solanum lycopersicum NCED by a series of aryl hydroxamic acid inhibitors is illustrated, and inhibitors D2 and D4 are assayed against NCED isozymes from Zea mays.

Keywords: 9'-cis-neoxanthin; Abscisic acid biosynthesis; Carotenoid cleavage dioxygenase; NCED.

MeSH terms

  • Dioxygenases* / antagonists & inhibitors
  • Dioxygenases* / metabolism
  • Enzyme Assays / methods
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology
  • Hydroxamic Acids / chemistry
  • Hydroxamic Acids / pharmacology
  • Plant Proteins / metabolism
  • Solanum lycopersicum* / drug effects
  • Solanum lycopersicum* / enzymology
  • Zea mays*

Substances

  • Dioxygenases
  • 9-cis-epoxy-carotenoid dioxygenase
  • Enzyme Inhibitors
  • Plant Proteins
  • Hydroxamic Acids