Lily bulb polyphenol oxidase obtained via an optimized multi-stage separation strategy for structural analysis and browning mechanism elucidation.

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  • Additional Information
    • Source:
      Publisher: Elsevier Applied Science Publishers Country of Publication: England NLM ID: 7702639 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-7072 (Electronic) Linking ISSN: 03088146 NLM ISO Abbreviation: Food Chem Subsets: MEDLINE
    • Publication Information:
      Publication: Barking : Elsevier Applied Science Publishers
      Original Publication: Barking, Eng., Applied Science Publishers.
    • Subject Terms:
      Catechol Oxidase*/chemistry ; Catechol Oxidase*/isolation & purification ; Catechol Oxidase*/metabolism ; Plant Proteins*/chemistry ; Plant Proteins*/isolation & purification ; Plant Proteins*/metabolism ; Molecular Docking Simulation*; Molecular Weight ; Maillard Reaction ; Plant Roots/chemistry ; Plant Roots/enzymology
    • Abstract:
      An optimized multi-stage separation strategy was developed to purify lily bulb polyphenol oxidase (PPO) for revealing its molecular structure. The PPO was purified 14.64-fold with high specific activity of 153,900 U/mg via optimized conditions of phosphate buffer pH (6.5), solid-liquid ratio (1:3), PVPP content (2 %), extraction time (4 h), followed by 30 %-50 % ammonium sulfate, diethylaminoethyl ion-exchange chromatography (0.1 M NaCl), and size exclusion chromatography. The PPO was identified as a dimeric protein with molecular weight of 135 kDa, containing 58.79 % random coil, 20.78 % α-helix, 17.41 % β-folding, and 3.02 % β-corner. The three-dimensional structure via homology modeling suggested that active center CuA bound to His151, His172, and His181, CuB bound to His307, His311, and His341. Furthermore, molecular docking indicated that its Phe337 and Tyr312 residues were catalytic cavity gates of catechol and 4-methylcatechol, respectively. Therefore, this study successfully analyzed purified PPO structure and further provided a theoretical foundation for its browning mechanism.
      Competing Interests: Declaration of competing interest No conflict of interest exists in the submission of this manuscript, and the manuscript is approved by all authors for publication. The work described was original research that has not been published previously, and not under consideration for publication elsewhere. All the authors listed have approved the manuscript that is enclosed.
      (Copyright © 2024 Elsevier Ltd. All rights reserved.)
    • Contributed Indexing:
      Keywords: Binding sites; Lily bulb; Molecular docking; Polyphenol oxidase; Purification optimization
    • Accession Number:
      EC 1.10.3.1 (Catechol Oxidase)
      0 (Plant Proteins)
    • Publication Date:
      Date Created: 20241020 Date Completed: 20241113 Latest Revision: 20241113
    • Publication Date:
      20241114
    • Accession Number:
      10.1016/j.foodchem.2024.141418
    • Accession Number:
      39427459