Inhibition mechanism: Phytic acid, NADH as a peroxidase inhibitor.

Peroxidase, a key enzyme causing enzymatic browning, and affected the potential values of fruit and vegetables. Phytic acid and NADH inhibited peroxidase in a competitive manner due to their reducing properties, and it's IC50 (1.18 ± 0.32) × 10-8, (8.02 ± 0.45) × 10-6 mol L-1, respectively. The interaction between phytic acid, NADH and peroxidase contributed to intrinsic fluorescence quenching and conformation alternation with a accuracy determination by multispectroscopic techniques (fluorescence spectra, FT-IR and CD spectra), respectively. Molecular docking simulation revealed that phytic acid, NADH interacted with His170, Ala34, Arg38, Ser73, Arg31, Lys174, Gln176, Asn175, Arg75; Gln176, Asn175, Phe221, Lys174, Gly173, Ser167, Phe172, Gly169, His170 in peroxidase, respectively and blocked substrates into catalytic reactions.

The activation mechanism of peroxidase by ultrasound.

The activation mechanism of peroxidase by ultrasound was investigated. The catalysis performance of peroxidase with ultrasound treatment was prior to the controls determined by UV-visible spectra and Fourier transform infrared spectra. The transformation of tryptophan residues in peroxidase led to the increase of a-helix and anti-parallel content in the secondary structure, and the content of p-sheet, p-turn and random coil in the secondary structure. In addition, under the atomic force microscope, under ultrasonic treatment, the large molecular clusters of tyrosinase are broken down into small molecular clusters. The current results showed that the activity of peroxidase is activated under ultrasonic treatment, which is mainly caused by ultrasound without conformational change, the catalytic center is exposed, and the affinity with the substrate is stronger.

The inhibition mechanism of luteolin on peroxidase based on multispectroscopic techniques.

Luteolin, a plant-derived flavonoid, was found to exert effective inhibitory effect to peroxidase activity in a non-competitive manner with an IC50 of (6.62 ± 0.45) × 10-5 mol L-1. The interaction between luteolin and peroxidase induced the formation of a static complex with a binding constant (Ksv) of 7.31 × 103 L mol-1 s-1 driven by hydrogen bond and hydrophobic interaction. Further, the molecular interaction between luteolin and peroxidase resulted in intrinsic fluorescence quenching, structural and conformational alternations which were determined by multispectroscopic techniques combined with computational molecular docking. Molecular docking results revealed that luteolin bound to peroxidase and interacted with relevant amino acid residues in the hydrophobic pocket. These results will provide information for screening additional peroxidase inhibitors and provide evidence of luteolin's potential application in preservation and processing of fruit and vegetables and clinical disease remedy.

Purification, kinetic parameters, and isoforms of polyphenol oxidase from "Xushu 22" sweet potato skin.

We purified and compared the polyphenol oxidase (PPO) isoenzymes present in "Xushu 22," a sweet potato. A membrane-bound form (mPPO) and two soluble forms (sPPO1 and sPPO2) were identified and purified using ammonium sulphate precipitation, ion exchange chromatography, gel filtration chromatography, and mass spectrometer. The three PPO isoforms were characterized enzymatically. The specific activity of mPPO was significantly higher than that of the two sPPO isoforms, being 24.55- and 13.89-fold higher than sPPO1 and sPPO2. The preferred substrates for mPPO and the two sPPOs were catechol and chlorogenic acid, respectively. They can be efficiently and safely applied to phenolic wastewater treatment after being immobilized. Both mPPO and the two sPPOs were rapidly inactivated under acid or base conditions and were unstable at 65°C. The most effective inhibitors of mPPO, sPPO1, and sPPO2 were glutathione, ascorbic acid, and L-cysteine, respectively. PRACTICAL APPLICATIONS: According to incomplete statistics, about 5% of sweet potatoes in china are wasted due to enzymatic browning every year. PPO was regarded as a key molecule contributing to enzymatic browning in fruits and vegetables during ripening, processing, and storage and responsible for economic and industrial loss. It's great importance to characterize the PPO from "Xushu 22" sweet potato and provide data on its inactivation. The three PPO isoforms were purified and identified by chromatography and mass spectrometer. This study will provide useful informations to have a better command of PPO from "Xushu 22" sweet potato and give ideals to solve the browning of sweet potato.