Experimental and Modelling Study of Enzymatic Oxidation of 5-o-Caffeoylquinic Acid by Polyphenol Oxidases

Abstract

A set of experiments to investigate the enzymatic oxidation of 5-o-caffeoylquinic acid with polyphenol oxidases was carried out in the temperature range from 18 to 39 °C. The kinetic data were obtained in a batch isothermal reactor made of quartz and operated at an optimum pH (approx. 7.0) for enzyme activity, using citrate-phosphate buffer. The decay of 5-o-caffeoylquinic acid concentrations was experimentally monitored with a spectrophotometer at 323 nm. A coefficient of molar absorptivity equal to 1.96·103 mM–1m–1 was found for 5-o-caffeoylquinic acid, based on absorbance measurements with standard aqueous solutions. The kinetic experimental results were used to calculate the parameters Km and vmax of the Michaelis-Menten model, which were 0.24 mM and 2.77·10–4 mM/s at 25 °C, respectively. The effects of temperature on the maximum velocity of oxidation of 5-o-caffeoylquinic acid and on the rate of enzyme deactivation were well described using the Arrhenius equation.

Keywords

• chlorogenic acids
• 5-CQA
• enzymatic oxidation
• polyphenol oxidases