Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>)
Currently, little is known about the characteristics of polyphenol oxidase from wheat bran, which is closely linked to the browning of wheat product. The wheat PPO was purified by ammonium sulfate precipitation, DEAE-Sepharose ion-exchange column, and Superdex G-75 chromatography column. Purified wh...
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MDPI AG
2024-03-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/29/6/1334 |
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| author | Kun Yu Wei He Xiaoli Ma Qi Zhang Chunxu Chen Peiyan Li Di Wu |
| author_facet | Kun Yu Wei He Xiaoli Ma Qi Zhang Chunxu Chen Peiyan Li Di Wu |
| author_sort | Kun Yu |
| collection | DOAJ |
| description | Currently, little is known about the characteristics of polyphenol oxidase from wheat bran, which is closely linked to the browning of wheat product. The wheat PPO was purified by ammonium sulfate precipitation, DEAE-Sepharose ion-exchange column, and Superdex G-75 chromatography column. Purified wheat PPO activity was 11.05-fold higher, its specific activity was 1365.12 U/mg, and its yield was 8.46%. SDS-PAGE showed that the molecular weight of wheat PPO was approximately 21 kDa. Its optimal pH and temperature were 6.5 and 35 °C for catechol as substrate, respectively. Twelve phenolic substrates from wheat and green tea were used for analyzing the substrate specificity. Wheat PPO showed the highest affinity to catechol due to its maximum V<sub>max</sub> (517.55 U·mL<sup>−1</sup>·min<sup>−1</sup>) and low K<sub>m</sub> (6.36 mM) values. Docking analysis revealed strong affinities between catechol, gallic acid, EGCG, and EC with binding energies of −5.28 kcal/mol, −4.65 kcal/mol, −4.21 kcal/mol, and −5.62 kcal/mol, respectively, for PPO. Sodium sulfite, ascorbic acid, and sodium bisulfite dramatically inhibited wheat PPO activity. Cu<sup>2+</sup> and Ca<sup>2+</sup> at 10 mM were considered potent activators and inhibitors for wheat PPO, respectively. This report provides a theoretical basis for controlling the enzymatic browning of wheat products fortified with green tea. |
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| last_indexed | 2024-04-24T17:57:55Z |
| publishDate | 2024-03-01 |
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| spelling | doaj.art-0e4092c88d9a45909e8d0c57d78dcee62024-03-27T13:57:05ZengMDPI AGMolecules1420-30492024-03-01296133410.3390/molecules29061334Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>)Kun Yu0Wei He1Xiaoli Ma2Qi Zhang3Chunxu Chen4Peiyan Li5Di Wu6School of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaSchool of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaSchool of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaSchool of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaSchool of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaSchool of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaSchool of Food Engineering, Anhui Science and Technology University, Chuzhou 233100, ChinaCurrently, little is known about the characteristics of polyphenol oxidase from wheat bran, which is closely linked to the browning of wheat product. The wheat PPO was purified by ammonium sulfate precipitation, DEAE-Sepharose ion-exchange column, and Superdex G-75 chromatography column. Purified wheat PPO activity was 11.05-fold higher, its specific activity was 1365.12 U/mg, and its yield was 8.46%. SDS-PAGE showed that the molecular weight of wheat PPO was approximately 21 kDa. Its optimal pH and temperature were 6.5 and 35 °C for catechol as substrate, respectively. Twelve phenolic substrates from wheat and green tea were used for analyzing the substrate specificity. Wheat PPO showed the highest affinity to catechol due to its maximum V<sub>max</sub> (517.55 U·mL<sup>−1</sup>·min<sup>−1</sup>) and low K<sub>m</sub> (6.36 mM) values. Docking analysis revealed strong affinities between catechol, gallic acid, EGCG, and EC with binding energies of −5.28 kcal/mol, −4.65 kcal/mol, −4.21 kcal/mol, and −5.62 kcal/mol, respectively, for PPO. Sodium sulfite, ascorbic acid, and sodium bisulfite dramatically inhibited wheat PPO activity. Cu<sup>2+</sup> and Ca<sup>2+</sup> at 10 mM were considered potent activators and inhibitors for wheat PPO, respectively. This report provides a theoretical basis for controlling the enzymatic browning of wheat products fortified with green tea.https://www.mdpi.com/1420-3049/29/6/1334wheat polyphenol oxidasepurificationbiochemical characterization |
| spellingShingle | Kun Yu Wei He Xiaoli Ma Qi Zhang Chunxu Chen Peiyan Li Di Wu Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>) Molecules wheat polyphenol oxidase purification biochemical characterization |
| title | Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>) |
| title_full | Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>) |
| title_fullStr | Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>) |
| title_full_unstemmed | Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>) |
| title_short | Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (<i>Wan grano</i>) |
| title_sort | purification and biochemical characterization of polyphenol oxidase extracted from wheat bran i wan grano i |
| topic | wheat polyphenol oxidase purification biochemical characterization |
| url | https://www.mdpi.com/1420-3049/29/6/1334 |
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