Comprehensive in vitro and in silico analysis of chokeberry (<i>Aronia melanocarpa</i>) polyphenol oxidase: purification and characterization of the enzyme and its inhibition by carbamate derivatives

Enzymatic browning, catalyzed by polyphenol oxidase, occurs due to cell membrane damage resulting from postharvest senescence and biotic and abiotic stressors, including physical injury, which leads to colour changes in fruits and vegetables. Chokeberry (Aronia melanocarpa), a rich source of polyphenols, has garnered significant attention in nutrition and medicine for its potent antioxidant capacity and potential protective effects on human health. This study aims to purify the polyphenol oxidase enzyme (PPO) from chokeberry, characterize its key biochemical properties, and evaluate the inhibitory activities and antioxidant capacity of carbamate derivatives. First, the enzyme was purified 51.94 times using single-step affinity chromatography. The enzyme's molecular weight was determined to be approximately 100 kDa through SDS-PAGE, appearing as a single band. The KM and Vmax values were found to be 5.43 mM and 1,428.5 EU/mL for catechol and 7.5 mM and 1,666.6 EU/mL for 4-methyl catechol. In inhibition studies, compound 7, which contains a carbamate group at the benzyl position and an electron-donating methyl group, emerged as the most potent PPO inhibitor with a Ki of 0.018 mu M. Additionally, compound 7 exhibited the strongest antioxidant activity. In silico molecular docking and binding energy calculations further supported these findings, revealing strong interactions between compound 7 and key active site residues. Induced-fit docking simulations demonstrated that compound 7 formed stable hydrogen bonds and hydrophobic interactions, consistent with its high binding affinity and low IC50 value. These computational results corroborate the experimental data and highlight the potential of rationally designed carbamate derivatives as effective PPO inhibitors.

Dergi

JOURNAL OF FOOD MEASUREMENT AND CHARACTERIZATION

Yıl

2026

Cilt

20

Sayı

1

Sayfa

413-425

ISSN

2193-4126

eISSN

2193-4134

DOI

10.1007/s11694-025-03718-4

WoS UID

WOS:001597938600001