Phenylalanine ammonia-lyase, flavanone 3β-hydroxylase and flavonol synthase enzyme activity by a new in vitro assay method in berry fruits

Food Chem. 2014 Jun 15:153:130-3. doi: 10.1016/j.foodchem.2013.12.034. Epub 2013 Dec 12.

Abstract

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.

Keywords: Berry; Enzyme activity; Flavanone 3β-hydroxylase; Flavonol synthase; Phenylalanine ammonia-lyase.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Enzyme Assays / methods*
  • Fruit / chemistry
  • Fruit / enzymology*
  • Mixed Function Oxygenases / analysis*
  • Oxidoreductases
  • Phenylalanine Ammonia-Lyase / analysis*
  • Plant Proteins / analysis*
  • Rosaceae / chemistry
  • Rosaceae / enzymology*

Substances

  • Plant Proteins
  • Mixed Function Oxygenases
  • Oxidoreductases
  • flavanone 3-dioxygenase
  • flavonol synthase
  • Phenylalanine Ammonia-Lyase