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Anti-polyphenol oxidase properties of total flavonoids from young loquat fruits: inhibitory activity and mechanism.

ABSTRACT: This study investigated anti-polyphenol oxidase activity and mechanism of purified total flavonoids (PTF) from young loquat fruits. PTF remarkably inhibited the activity of polyphenol oxidase (PPO) with an IC ₅₀ value of 21.03 ± 2.37 μg/mL. Based on enzyme kinetics, PTF was found to be a potent, mixed-type, and reversible inhibitor of PPO. The fluorescence intensity of PPO was quenched by PTF through forming a PTF-PPO complex in a static procedure. Therefore, this study authenticated PTF as an efficient PPO inhibitor, which would contribute to their utilization in food industry.

SUBMITTER: Huang GL

PROVIDER: S-EPMC8806263 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Anti-polyphenol oxidase properties of total flavonoids from young loquat fruits: inhibitory activity and mechanism.

Huang Gui-Li GL Sun Ling-Xiang LX Ma Jia-Jia JJ Sui Si-Yao SY Wang Yu-Ning YN

Bioengineered 20211201 1

This study investigated anti-polyphenol oxidase activity and mechanism of purified total flavonoids (PTF) from young loquat fruits. PTF remarkably inhibited the activity of polyphenol oxidase (PPO) with an <i>IC</i><sub>50</sub> value of 21.03 ± 2.37 μg/mL. Based on enzyme kinetics, PTF was found to be a potent, mixed-type, and reversible inhibitor of PPO. The fluorescence intensity of PPO was quenched by PTF through forming a PTF-PPO complex in a static procedure. Therefore, this study authenti ...[more]

PMID: 33587004

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Project description:Polyphenols are naturally occurring compounds found largely in fruits and vegetables. The antioxidant properties of these polyphenols including total phenolic content (TPC), total flavonoid content (TFC), tannin content, 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH), 2,2'-azinobis-(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS) scavenging abilities and ferric ion reducing antioxidant power (FRAP) were measured among sixteen (16) plant foods (mango, blueberry, strawberry, black carrot, raspberry, dark grapes, garlic, ginger, onion, cherry, plum, apple, papaya, peach, pear and apricot) by modifying, standardising and translating existing antioxidant methods using a 96-well plate reader. Eighteen targeted phenolic acids and flavonoids were characterised and quantified using high-performance liquid chromatography-photometric diode array (HPLC-PDA) and verified by modifying an existing method of liquid chromatography coupled with electrospray-ionisation triple quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS). While most of these compounds were accurately detected by the HPLC-PDA at a low concentration, a few polyphenols in low concentrations could be only be characterised using the LC-ESI-QTOF/MS method. Our results showed that mango possessed the highest overall antioxidant activity, phenolic acid and flavonoid content among the selected fruits. Factor analysis (FA) and Pearson's correlation tests showed high correlations among ABTS, DPPH, FRAP and phenolic acids, implying the comparable capabilities of scavenging the DPPH/ABTS free radicals and reducing ferric ions from the antioxidant compounds in the samples. Phenolic acids contributed significantly to the antioxidant activities, and flavonoids contributed more to tannin content based on the correlations. Overall, methods modified and standardized in this study can provide better understanding of high throughput technologies and increase the reliability of antioxidant data of different plant foods.

Project description:In this study, a polydopamine (PDA)-modified hollow fiber-immobilized xanthine oxidase (XOD) was prepared for screening potential XOD inhibitors from flavonoids. Several parameters for the preparation of PDA-modified hollow fiber-immobilized XOD, including the dopamine concentration, modification time, XOD concentration and immobilization time, were optimized. The results show that the optimal conditions for immobilized XOD activity were a dopamine concentration of 2.0 mg/mL in 10.0 mM Tris-HCl buffer (pH 8.5), a modification time of 3.0 h, an XOD concentration of 1000 μg/mL in 10.0 mM phosphate buffer (pH 7.5) and an immobilization time of 3.0 h. Subsequently, the enzymatic reaction conditions such as the pH value and temperature were investigated, and the enzyme kinetics and inhibition parameters were determined. The results indicate that the optimal pH value (7.5) and temperature (37 °C) of the PDA-modified hollow fiber-immobilized XOD were consistent with the free enzyme. Moreover, the PDA-modified hollow fiber-immobilized XOD could still maintain above 50% of its initial immobilized enzyme activity after seven consecutive cycles. The Michaelis-Menten constant (Km) and the half-maximal inhibitory concentration (IC50) of allopurinol on the immobilized XOD were determined as 0.25 mM and 23.2 μM, respectively. Furthermore, the PDA-modified hollow fiber-immobilized XOD was successfully applied to evaluate the inhibitory activity of eight flavonoids. Quercetin, apigenin, puerarin and epigallocatechin showed a good inhibition effect, and their percentages of inhibition were (79.86 ± 3.50)%, (80.98 ± 0.64)%, (61.15 ± 6.26)% and (54.92 ± 0.41)%, respectively. Finally, molecular docking analysis further verified that these four active compounds could bind to the amino acid residues in the XOD active site. In summary, the PDA-modified hollow fiber-immobilized XOD is an efficient method for the primary screening of XOD inhibitors from natural products.

Dataset Information

Anti-polyphenol oxidase properties of total flavonoids from young loquat fruits: inhibitory activity and mechanism.

Publications

Anti-polyphenol oxidase properties of total flavonoids from young loquat fruits: inhibitory activity and mechanism.

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