Open Access Research article

CYP98A22, a phenolic ester 3’-hydroxylase specialized in the synthesis of chlorogenic acid, as a new tool for enhancing the furanocoumarin concentration in Ruta graveolens

Fazeelat Karamat12, Alexandre Olry12, Sébastien Doerper12, Guilhem Vialart12, Pascaline Ullmann3, Danièle Werck-Reichhart3, Frédéric Bourgaud12 and Alain Hehn12*

Author Affiliations

1 Université de Lorraine UMR 1121, Agronomie et Environnement Nancy-Colmar, ENSAIA 2 avenue de la forêt de Haye, 54505, Vandœuvre-lès-Nancy, France

2 INRA UMR 1121, Agronomie et Environnement Nancy-Colmar, ENSAIA 2 avenue de la forêt de Haye, 54505, Vandœuvre-lès-Nancy, France

3 CNRS UPR 2357 28 rue Goethe, Strasbourg, 67000, France

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BMC Plant Biology 2012, 12:152  doi:10.1186/1471-2229-12-152

Published: 29 August 2012

Additional files

Additional file 1:

A) Alignment done on 9 CYP98A (CYP98A1 (AF029856) ; CYP98A2 (AF022458); CYP98A3 (AC002409); CYP98A6 (AB017418); CYP98A8 (AC011765); CYP98A9(AC011765); CYP98A13 (AAL99200); CYP98A19 (AY064170); CYP98A20 (AY065995). Only the C-terminal ends of the proteins are represented. The conserved sequences used to design the CODEHOP primers are highlighted in yellow. B) Nucleotidic and peptidic sequence of CYP98A22 (Genbank JF799117).

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Additional file 2:

Identification of the metabolization product of p-coumaroyl quinate. A) MS analysis of the metabolization product: B) MS analysis of caffeoyl quinate; C) MS2 analysis of the metabolization product: D) MS2 analysis of caffeoyl quinate.

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Additional file 3:

Identification of the metabolization product of p-coumaroyl shikimate. A) MS analysis of the metabolization product: B) MS analysis of caffeoyl shikimate; C) MS2 analysis of the metabolization product: D) MS2 analysis of caffeoyl shikimate.

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Additional file 4:

Measurement of apparent Kmof CYP98A22. Reaction rates versus different concentrations of p-coumaroyl shikimate (4μM-50μM) (A) and of p-coumaroyl quinate(1μM-30 μM) (B). Apparent Km values were measured by fitting the data to the Michaelis-Menten equation using the software SigmaPLOT 12.0.

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