title: High Hydrostatic Pressure-Assisted Enzymatic Treatment Improves Antioxidant and Anti-inflammatory Properties of Phosvitin.
abstract: BACKGROUND
  Phosvitin (PV) is a highly-phosphorylated metal-binding protein in egg yolk. Phosphoserine clusters make PV resistant to enzymatic digestion, which might be nutritionally undesirable.
  
  
  OBJECTIVE
  This study was designed to determine the effects of high hydrostatic pressure and enzymatic hydrolysis (HHP-EH) on the antioxidant and anti-inflammatory properties of PV hydrolysates (PVHs).
  
  
  METHODS
  PV was hydrolyzed by alcalase, elastase, savinase, thermolysin, and trypsin at 0.1, 50, and 100 MPa pressure levels. PVHs were evaluated for degree of hydrolysis, molecular weight distribution patterns, antioxidant and anti-inflammatory properties in chemical and cellular models. The effect of PVH on gene expression of pro-inflammatory cytokines (TNF-α and IL-1β) was also evaluated using real time-PCR. The hydrolysate with most potent antioxidant and anti-inflammatory properties was subjected to LC-MS/MS analysis to identify the peptide sequence.
  
  
  RESULTS
  Hydrolysates produced at 100 MPa exhibited higher degree of hydrolysis and greater reducing power and free radical scavenging activity compared to those obtained at atmospheric pressure. After adjusting the phosphate content, alcalase- and trypsin-digested PVHs showed superior iron chelation capacity (69-73%), regardless of pressure. Both alcalase- and trypsin-digested PVHs significantly inhibited nitric oxide production by RAW264.7 macrophage cells. LPS-stimulated up-regulation of proinflammatory cytokines was also suppressed by alcalase-digested PVH.
  
  
  CONCLUSIONS
  The HHP-EH method could play a promising role in the production of bioactive peptides from hydrolysis-resistant proteins. HHP-assisted PVH may be useful in preparing a potential pharmaceutical with antioxidant and anti-inflammatory properties.
authors.forename: Heejoo
authors.initials: H
authors.lastname: Yoo
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Bamdad
authors.forename: Naiyana
authors.initials: N
authors.lastname: Gujral
authors.forename: Joo-Won
authors.initials: JW
authors.lastname: Suh
authors.affiliation: Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 114 St-89 Ave., Edmonton, Alberta, T6G 2H7, Canada.
authors.forename: Hoon
authors.initials: H
authors.lastname: Sunwoo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D006868:Hydrolysis; D006874:Hydrostatic Pressure; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology; D010774:Phosvitin / Q000737:chemistry / Q000494:pharmacology*; D011492:Protein Hydrolysates / Q000302:isolation & purification / Q000494:pharmacology*; D012697:Serine Endopeptidases / Q000737:chemistry*
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D010455:Peptides; D011492:Protein Hydrolysates; D010774:Phosvitin; D012697:Serine Endopeptidases
keywords: High hydrostatic pressure; antioxidant; hydrolysis; nitric oxide; phosvitin; pro-inflammatory cytokines