Phenylalanine ammonia-lyase, flavanone 3β-hydroxylase and flavonol synthase enzyme activity by a new in vitro assay method in berry fruits.
An HPLC method for the determination of phenylalanine ammonia-lyase, flavanone 3β-hydroxylase and flavonol synthase enzyme activity is proposed. This method is based on the determination of the compounds produced and consumed on the enzymatic reaction in just one chromatographic analysis. Optimisation of the method considered kinetic studies to establish the incubation time to perform the assay. The method here described proved to be an interesting approach to measure the activities of the three enzymes simultaneously increasing the rapidity, selectivity and sensitivity over other exiting methods. The enzyme activity method developed was applied to strawberry, raspberry, blackberry, redcurrant and blackcurrant fruits.
Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN-CSIC), Consejo Superior de Investigaciones Científicas (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain. Electronic address: gemaflores@ictan.csic.es.
Gema
G
Flores
Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN-CSIC), Consejo Superior de Investigaciones Científicas (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain.
Fernando
F
De la Peña Moreno
Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN-CSIC), Consejo Superior de Investigaciones Científicas (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain.
Gracia Patricia
GP
Blanch
Instituto de Ciencia y Tecnología de Alimentos y Nutrición (ICTAN-CSIC), Consejo Superior de Investigaciones Científicas (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain.
Maria Luisa Ruiz
ML
Del Castillo
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D005638:Fruit / Q000737:chemistry / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000032:analysis*; D010088:Oxidoreductases; D010650:Phenylalanine Ammonia-Lyase / Q000032:analysis*; D010940:Plant Proteins / Q000032:analysis*; D027824:Rosaceae / Q000737:chemistry / Q000201:enzymology*
D010940:Plant Proteins; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; C048797:flavanone 3-dioxygenase; C085313:flavonol synthase; D010650:Phenylalanine Ammonia-Lyase
Berry; Enzyme activity; Flavanone 3β-hydroxylase; Flavonol synthase; Phenylalanine ammonia-lyase