Different types of microsomal enzymes catalyzeortho- orpara-hydroxylation in the biosynthesis of carnation phytoalexins
- 2 December 1991
- journal article
- Published by Wiley in FEBS Letters
- Vol. 294 (1-2), 67-72
- https://doi.org/10.1016/0014-5793(91)81345-9
Abstract
Cell suspension cultures of carnation (Dianthus caryophyllus L.) accumulate, upon challenge with crude fungal elicitor, various dianthramide phytoalexins, all of which derive from N-benzoylanthranilate. In vitro, microsomes from the elicited carnation cells hydroxylated N-benzoylanthranilate in the 4- and/or 2′-positions to yield the hydroxyanthranilate and/or salicyloyl derivatives. 2′-Hydroxylation was shown to precede 4-hydroxylation in the formation of N-salicyloyl-4-hydroxyanthranilate, and both these activities depended strictly on NADPH and molecular oxygen. 4-Hydroxylation was shown to be catalyzed by cytochrome P-450-dependent monooxygenase(s), whereas the 2′-hydroxylating activity appeared to be due to a novel class of enzymes, also responding synergistically to NADH in combination with NADPH and showing apparent inhibition by cytochrome c but not by carbon monoxide. The difference in type of 4- and 2′-hydroxylases was corroborated by the exclusive inhibition of either activity in imidazole vs. MOPS buffers as well as their differential heat sensitivities. In the course of these studies, low concentrations of N-salicyloylanthranilate turned out to inhibit the cytochrome P-450-dependent 4-hydroxylation more strongly than any of the commercial inhibitor chemicals tested, while neither the substrate, N-benzoylanthranilate, nor the final product, N-salicyloyl-4-hydroxyanthranilate, exhibited such significant inhibition. In addition, 2′-hydroxylation activity was affected much less by N-benzoylanthranilate, N-salicyloylanthranilate or by inhibitor chemicals. The results demonstrate the requirement of two different classes of hydroxylase activities that appear to introduce the antimycotic quality to the dianthramides for phytoalexin defenseKeywords
This publication has 18 references indexed in Scilit:
- Novel synthesis of dianthalexin (phytoalexin) analogues preparationJournal of Heterocyclic Chemistry, 1988
- Elicitor‐induced biosynthesis of psoralens in Ammi majus L. suspension culturesEuropean Journal of Biochemistry, 1988
- Dianthramides (N-benzoyl and N-paracoumarylanthranilic acid derivatives) from elicited tissues of Dianthus caryophyllusPhytochemistry, 1988
- Microsomal isoflavone 2′‐ and 3′‐hydroxylases from chickpea (Cicer arietinum L.) cell suspensions induced for pterocarpan phytoalexin formationFEBS Letters, 1987
- Dianthramides A and B, two N-benzoylanthranilic acid derivatives from elicited tissues of Dianthus caryophyllusPhytochemistry, 1984
- Induction and Characterization of a Microsomal Flavonoid 3′‐Hydroxylase from Parsley Cell CulturesEuropean Journal of Biochemistry, 1983
- Metabolisme des composes phenoliques chez le petunia v. utilisation de la phenylalanine par des chloroplastes isolesPlant Science Letters, 1977
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Zur Frage der ortho-Hydroxylierung aromatischer Carbonsäuren in höheren PflanzenHoppe-Seyler´s Zeitschrift Für Physiologische Chemie, 1971
- Uncoupling of oxygen activation from hydroxylation in a bacterial salicylate hydroxylaseBiochemical and Biophysical Research Communications, 1970