BACKGROUND Food and agricultural wastes are produced in huge amounts yearly, putting extra effort into their removal or valorization. One of these wastes is the zucchini heads (ZH) which, in this study, was used as a source to isolate peroxidase using three-phase portioning (TPP). RESULTS Different parameters of TPP were optimized to ensure obtaining the maximal activity recovery and purity of the enzyme. The purity of the isolated enzyme was performed using the protein homogeneity module of dynamic light scattering. This was followed by the determination of the optimal pH and temperature of the isolated ZH peroxidase. It was found that sodium citrate at a concentration of 15%, pH 8, 1-butanol as the upper alcoholic phase, and an alcohol/crude extract ratio of 0.

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1 were the best conditions for ZH peroxidase isolation. The obtained activity recovery and purification fold were 159% and 10.05, respectively. The isolated ZH peroxidase displayed a high purity as emphasized via dynamic light scattering. The optimum pH and temperature were 8 and 25 C. CONCLUSIONS The present study was the first to isolate and purify peroxidase from ZH using TPP in one step