title: Isolation and characterization of anti-inflammatory peptides derived from whey protein.
abstract: The present study was conducted to isolate and characterize anti-inflammatory peptides from whey protein hydrolysates using alcalase. Nine subfractions were obtained after sequential purification by ultrafiltration, Sephadex G-25 gel (GE Healthcare, Uppsala, Sweden) filtration chromatography, and preparative HPLC. Among them, subfraction F4e showed the strongest inhibitory activity on interleukin-1β (IL-1β), cyclooxygenase-2, and tumor necrosis factor-α (TNF-α) mRNA expression in lipopolysaccharide-induced RAW 264.7 mouse macrophages. Eight peptides, including 2 new peptides-Asp-Tyr-Lys-Lys-Tyr (DYKKY) and Asp-Gln-Trp-Leu (DQWL)-were identified from subfractions F4c and F4e, respectively, using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. Peptide DQWL showed the strongest inhibitory ability on IL-1β, cyclooxygenase-2, and TNF-α mRNA expression and production of IL-1β and TNF-α proteins at concentrations of 10 and 100μg/mL, respectively. Additionally, DQWL treatment significantly inhibited nuclear factor-κB activation by suppressing nuclear translocation of nuclear factor-κB p65 and blocking inhibitor κB kinase phosphorylation and inhibitor κB degradation together with p38 mitogen-activated protein kinase activation. Our study suggests that peptide DQWL has anti-inflammatory potential; further confirmation using an in vivo model is needed.
authors.affiliation: Institute of Food and Nutraceutical Science, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology & Business University, Beijing 100048, China. Electronic address: liujiefantasy@163.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.affiliation: Institute of Food and Nutraceutical Science, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: hmshi@sjtu.edu.cn.
authors.forename: Haiming
authors.initials: H
authors.lastname: Shi
authors.affiliation: Department of Nutrition and Food Science, University of Maryland, College Park 20742.
authors.forename: Liangli Lucy
authors.initials: LL
authors.lastname: Yu
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000302:isolation & purification* / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D004151:Dipeptides / Q000302:isolation & purification / Q000494:pharmacology; D053583:Interleukin-1beta / Q000378:metabolism; D008070:Lipopolysaccharides / Q000633:toxicity; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology; D010766:Phosphorylation; D000067996:RAW 264.7 Cells; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000067816:Whey Proteins / Q000737:chemistry*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D004151:Dipeptides; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010455:Peptides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D000067816:Whey Proteins; C509811:tryptophan-leucine; C042696:tyrosyltyrosine; C078779:aspartyltyrosine; C024575:lysyllysine; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; D048051:p38 Mitogen-Activated Protein Kinases
keywords: alcalase; anti-inflammatory peptide; purification; whey protein