Fennel seeds were recognized as a promising polyphenol oxidase (PPO) source upon investigating some edible green plants (carob, jujube, coriander, fennel, and licorice). The fennel PPO enzyme was purified by three-phase partitioning and biochemically characterized in detail for the first time. The purification fold and activity recovery values were determined as 20-fold and 120%, respectively. Its molecular weight was 27.8 kDa. The temperature for the selected substrates (catechol, 4-tert-butylcatechol, 4-methylcatechol, and pyrogallol) was 30 degrees C, while the optimum pH value varied from 5.0 to 7.0 depending on the substrate. The k(cat)/Km values exhibited that the enzyme presented the best activity towards catechol among the substrates used. Sodium metabisulfite, ascorbic acid, benzoic acid, L-cysteine, thiourea, beta-mercaptoethanol, and glutathione prominently inhibited PPO activity. A remarkable decrease in PPO activity was observed at elevated concentrations of organic solvents, but in cases of the solvents with polarity indexes >= 5.1, the residual activity maintained more than 75% of its original activity up to 10% (v/v). Consequently, the current study suggested that fennel seeds could be used in various industrial sectors to produce low-cost polyphenol oxidase enzymes with an agricultural origin. (C) 2020 Elsevier B.V. All rights reserved.