Purification and characterisation of polyphenol oxidase (PPO) from eggplant (Solanum melongena)

► Eggplant was found to be a very rich source of polyphenol oxidase (PPO). ► Native PPO was 259-fold purified using standard chromatographic procedures. ► The PPO was found to be a 112kDa homodimer. ► The enzyme showed very low Km and high catalytic efficiency with 4-methyl catechol. ► GRAS compound cysteine hydrochloride was found to be an excellent PPO inhibitor. Eggplant (Solanum melongena) is a very rich source of polyphenol oxidase (PPO), which negatively affects its quality upon cutting and postharvest processing due to enzymatic browning. PPO inhibitors, from natural or synthetic sources, are used to tackle this problem. One isoform of PPO was 259-fold purified using standard chromatographic procedures. The PPO was found to be a 112kDa homodimer. The enzyme showed very low Km (0.34mM) and high catalytic efficiency (3.3×106) with 4-methyl catechol. The substrate specificity was in the order: 4-methyl catechol>tert-butylcatechol>dihydrocaffeic acid>pyrocatechol. Cysteine hydrochloride, potassium metabilsulphite, ascorbic acid, erythorbic acid, resorcylic acid and kojic acid showed competitive inhibition, whereas, citric acid and sodium azide showed mixed inhibition of PPO activity. Cysteine hydrochloride was found to be an excellent inhibitor with the low inhibitor constant of 1.8μM.