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Characterization of cinnamate 4-hydroxylase (CYP73A) and p-coumaroyl 3'-hydroxylase (CYP98A) from Leucojum aestivum, a source of Amaryllidaceae alkaloids.
Karimzadegan, Vahid; Koirala, Manoj; Sobhanverdi, Sajjad; Merindol, Natacha; Majhi, Bharat Bhusan; Gélinas, Sarah-Eve; Timokhin, Vitaliy I; Ralph, John; Dastmalchi, Mehran; Desgagné-Penix, Isabel.
Afiliación
  • Karimzadegan V; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • Koirala M; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • Sobhanverdi S; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • Merindol N; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • Majhi BB; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • Gélinas SE; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
  • Timokhin VI; Department of Energy's Great Lakes Bioenergy Research Center, Wisconsin Energy Institute, Madison, WI, 53726, USA.
  • Ralph J; Department of Energy's Great Lakes Bioenergy Research Center, Wisconsin Energy Institute, Madison, WI, 53726, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA.
  • Dastmalchi M; Department of Plant Science, McGill University, Montréal, Québec, Canada.
  • Desgagné-Penix I; Department of Chemistry, Biochemistry and Physics, Université Du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada. Electronic address: Isabel.Desgagne-Penix@uqtr.ca.
Plant Physiol Biochem ; 210: 108612, 2024 May.
Article en En | MEDLINE | ID: mdl-38598867
ABSTRACT
Biosynthesis of Amaryllidaceae alkaloids (AA) starts with the condensation of tyramine with 3,4-dihydroxybenzaldehyde. The latter derives from the phenylpropanoid pathway that involves modifications of trans-cinnamic acid, p-coumaric acid, caffeic acid, and possibly 4-hydroxybenzaldehyde, all potentially catalyzed by hydroxylase enzymes. Leveraging bioinformatics, molecular biology techniques, and cell biology tools, this research identifies and characterizes key enzymes from the phenylpropanoid pathway in Leucojum aestivum. Notably, we focused our work on trans-cinnamate 4-hydroxylase (LaeC4H) and p-coumaroyl shikimate/quinate 3'-hydroxylase (LaeC3'H), two key cytochrome P450 enzymes, and on the ascorbate peroxidase/4-coumarate 3-hydroxylase (LaeAPX/C3H). Although LaeAPX/C3H consumed p-coumaric acid, it did not result in the production of caffeic acid. Yeasts expressing LaeC4H converted trans-cinnamate to p-coumaric acid, whereas LaeC3'H catalyzed specifically the 3-hydroxylation of p-coumaroyl shikimate, rather than of free p-coumaric acid or 4-hydroxybenzaldehyde. In vivo assays conducted in planta in this study provided further evidence for the contribution of these enzymes to the phenylpropanoid pathway. Both enzymes demonstrated typical endoplasmic reticulum membrane localization in Nicotiana benthamiana adding spatial context to their functions. Tissue-specific gene expression analysis revealed roots as hotspots for phenylpropanoid-related transcripts and bulbs as hubs for AA biosynthetic genes, aligning with the highest AAs concentration. This investigation adds valuable insights into the phenylpropanoid pathway within Amaryllidaceae, laying the foundation for the development of sustainable production platforms for AAs and other bioactive compounds with diverse applications.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Plantas / Alcaloides de Amaryllidaceae / Transcinamato 4-Monooxigenasa Idioma: En Revista: Plant Physiol Biochem Asunto de la revista: BIOQUIMICA / BOTANICA Año: 2024 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Francia
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Plantas / Alcaloides de Amaryllidaceae / Transcinamato 4-Monooxigenasa Idioma: En Revista: Plant Physiol Biochem Asunto de la revista: BIOQUIMICA / BOTANICA Año: 2024 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Francia