Purification of Polyphenol Oxidase from Potato and Investigation of the Inhibitory Effects of Phenolic Acids on Enzyme Activity

Abstract

Background: Polyphenol Oxidase (PPO) belongs to the oxidoreductase enzyme family. Methods: Here, PPO was purified from potato using Sepharose 4B-L-tyrosine-p-aminobenzoic acid affinity chromatography. It determined the interactions between some phenolic acids and the enzyme. Results: The enzyme was obtained with a specific activity of 15333.33 EU/mg protein and 7.87- fold purification. It was found that phenolic acids exhibited inhibitory properties for PPO. The IC50 values of the phenolic acids were found in the range of 0.36-2.12 mM, and their Ki values were found in the range of 0.28± 0.07-1.72±0.32 mM. It was determined that all studied compounds displayed a competitive inhibition effect. Among these compounds, 3-hydroxybenzoic acid was found to be the most effective PPO inhibitor (Ki: 0.28±0.07 mM). Conclusion: Investigating the inhibition kinetics of the enzyme will simplify the testing of PPO inhibitor candidates.

In vitro enzymatic synthesis of a monomeric theaflavin using a polyphenol oxidase isozyme from tea (Camellia sinensis) leaf

“…Different isolation and purification protocols have been reported, and the purification fold depends on the methods used, localisation of enzyme and species source (Panadare & Rathod, 2018). For example, PPO from borage was purified 3.59‐fold using the three‐phase partitioning purification technique (Alici & Arabaci, 2016); PPO from potato was purified 7.87‐fold using Sepharose 4B‐L‐tyrosine‐ p ‐aminobenzoic acid affinity chromatography (Bayrak et al ., 2019), and PPO from Tadela date fruit was purified 79.9‐fold using ammonium sulphate precipitation, Q‐Sepharose Big Beats method and HPLC zorbax chromatography (Benaceur et al ., 2019).…”

Prevention of Enzymatic Browning by Natural Extracts and Genome-Editing: A Review on Recent Progress

“…Therefore, the most attractive way to prevent food browning would be by natural methods. Seeking “natural” agents as food preservatives derived from plant extracts, essential oils, bioactive compounds, and purified secondary metabolites, has attracted food researchers’ attention [ 28 , 29 ].…”

Isolation of Some Phenolic Compounds from Plantago subulata L. and Determination of Their Antidiabetic, Anticholinesterase, Antiepileptic and Antioxidant Activity

“…Today, acetylcholinesterase (AChE) inhibitors are used in neurodegenerative disease therapy. They are based on which source from plants [24–26] . Herbs could effectively serve as origins for new therapeutics [27] .…”