Purification and characterization of catechol oxidase from Posof Badele apple (Malus domestica L): in vitro and in silico studies

Abstract In the current study, polyphenol oxidase (PPO), responsible for enzymatic browning in fruits, was purified from Posof Badele apple (PB) (Malus domestica L.) in two steps as acetone precipitation and affinity chromatography. After purification, the purity of PBPPO was checked by using SDS-PAGE. It was figured out that PBPPO had maximum activity at pH 6.0 and 10 °C and it was stable at pH 5.5 and temperatures of 0–30 °C. Besides, it was determined that Al3+ and Cu2+ metal ions activated the enzyme and the PBPPO was strongly inhibited by ascorbic acid with a Ki constant of 1.67 ± 0.35 µM. Inhibitor-enzyme interactions were examined by molecular docking studies and it was revealed that ascorbic acid had the lowest docking score of −6.54 kcal/mol. We wanted to draw attention to the PB apple, which is red inside as well as outside and very rich in terms of nutrient content.