Antarctic krill antioxidant peptides show inferior IgE-binding ability and RBL-2H3 cell degranulation

Enzymatic hydrolysis,isolation,and purification might make a great deal of difference in antioxidant activity and antigenicity of peptide components.This study aimed to isolate and purify antioxidant peptide components from Antarctic krill and evaluate their allergenicity of them.Electron paramagnetic resonance(EPR)spectroscopy results indicated 3-10 kDa Antarctic krill hydrolysates(AKHs)had higher DPPH and·OH radical scavenging rates.And the second component(N2-2)purified 3-10 kDa hydrolysate showed better ability to scavenge DPPH and·OH radicals(P<0.05),which were(47.43±2.18)%and(34.33±1.25)%,respectively.Additionally,indirect-ELISA results revealed that N2-1 had a weaker ability to bind specific IgE and that N2-2 had a lower binding capability to specific IgG1(P<0.05).And N2-2 had a higher EC50 value of(5.29±0.95)ng/mL(P<0.05)in cell degranulation assay,which was about 13.80 times that of Antarctic krill.Therefore,N2-2 might be the potential source of the antioxidant peptides with lower allergenicity.

Antioxidant peptides encrypted in flaxseed proteome:An in silico assessment

Flaxseed proteins and antioxidant peptides(AP)encrypted in their sequences were analysed in silico with a range of bioinformatics tools to study their physicochemical properties,allergenicity,and toxicity.Nine proteases(digestive,plant and microbial sources)were assessed for their ability to release known APs from 23 mature flaxseed storage proteins using the BIOPEP database.The families of proteins identified were predominantly globulins,oleosins,and small amount of conlinin.Overall,253 APs were identified from these proteins.More peptides were released by enzymatic hydrolysis from the globulins than those from oleosins and conlinin.Compared with other enzymes studied,the plant proteases(papain,ficin,and bromelain)were found to be superior to releasing APs from the flaxseed proteins.Analysis of toxicity by ToxinPred showed that none of the peptides released was toxic.Most of the APs showed structural features that are important for antioxidation,including relatively low molecular weight(dipeptides and tripeptides only);amphipathic properties(hydrophobicity range of-0.5 to+0.5);relatively low Boman index(≤2);broad range of pI(3.7-10.8),and an abundance of antioxidant amino acid residues(e.g.glutamic acid and histidine).This study demonstrate the suitability of flaxseed proteins as a source of APs.

Peptidomic analysis of whey protein hydrolysates and prediction of their antioxidant peptides

Enzymatic hydrolysis of proteins is a breakdown process of peptide bond in proteins,releasing some peptides with potential biological functions.Previous studies on enzymatic hydrolysis of whey proteins have not identified the complete peptide profiles after hydrolysis.In this study,we reconstructed a profile of peptides from whey hydrolysates with two enzymes and different processing conditions.We also developed an ensemble machine learning predictor to classify peptides obtained from whey hydrolysis.A total of 2572 peptides were identified over three process conditions with two enzymes in duplicate.499 peptides were classified and chosen as potential antioxidant peptides from whey proteins.The peptides classified as antioxidants in the hydrolysates had a proportion of 13.1%-24.5%regarding all peptides identified.These results facilitate the selection of promising peptides involved in the antioxidant properties during the enzymatic hydrolysis of whey proteins,aiding the discovery of novel antioxidant peptides.

The protective effects of peptides from Chinese baijiu on AAPH-induced oxidative stress in HepG2 cells via Nrf2 signaling pathway

Antioxidant peptides have been widely reported.However,only a few reports have been published examining the antioxidant peptides derived from Chinese baijiu.In this study,6 novel peptides derived from Chinese baijiu were identified successfully using high-performance liquid chromatography-quadrupoletime-of-flight mass spectrometry(HPLC-QTOF-MS)with a concentration of 0.835–24.540μg/L.The underlying molecular mechanisms were investigated,and their cytoprotective effects were examined against 2,2’-azobis(2-methylpropanimidamidine)dihydrochloride(AAPH)-induced oxidative stress in Hep G2 cells.The results showed that these peptides exerted protective effects by suppressing reactive oxygen species(ROS)generation,preventing malondialdehyde(MDA)formation,and upregulating cellular antioxidant enzyme activities(SOD,CAT,and GSH-Px)in a dose-dependent manner.Further experiments proved that these peptides exerted antioxidant effects via Nrf2/ARE-mediated signaling pathway by promoting Nrf2 nuclear translocation,inhibiting ubiquitination,and enhancing transcription capacity of Nrf2 in Hep G2 cells.These findings provide the molecular basis for the effects of antioxidant peptides derived from Chinese baijiu,which is important for a deeper understanding of the relationship between human health and moderate drinking.

In vitro Antioxidant Effects of Porphyra haitanensis Peptides on H_2O_2-Induced Damage in HepG2 Cells

In this study,protein from Porphyra haitanensis was used as raw material to prepare an antioxidant peptide,and its antioxidant activity was evaluated in vitro.A model of H_2O_2-induced oxidative damage in Hep G2 cells was established,and the effects of Porphyra haitanensis hydrolysates (PHHs) on superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected.Finally,the structure of PHHs was identified by ESI-MS/MS.The results showed that the 1,1-diphenyl-2-pyridylhydrazine(DPPH)-free radical-scavenging ability of PHHs was the strongest (59.28%at 1.0 mg m L~(-1)) when hydrolyzed with an acidic protease for 4 h.PHHs with different concentrations had protective effects on H_2O_2-induced damage to Hep G2 cells,and the protective effect was enhanced with increasing concentrations.When the level was 400μg m L~(-1),the cell survival rate was as high as 88.62%.Moreover,PHHs can significantly reduce oxidative damage to Hep G2 cells by H_2O_2,improve SOD activity,and reduce MDA content.The tetrapeptide Asp-Lys-Ser-Thr,with a molecular weight of 448 Da,was identified as an important fraction of PHHs by high-resolution mass spectrometry.

Production and Purification of Antioxidant Peptides from Flatfish Skin Protein Hydrolysates

Antioxidant peptides of flatfish skin protein hydrolyzed by four enzymes （Papain, Pepsin, Trypsin and Neutrase, respectively）were investigated. The Trypsin hydrotysate obtained by hydrolysis exhibited the highest 1,1- dipheny-l-2-picrylhydrazyl（DPPH） radical scavenging activity （DRSA） compared with other hydrolysates. Re- sponse surface method ology （RSM）, based on Box-Behnken design, was used to study the influence of hydrolysis conditions on the DRSA. The optimal hydrolysis conditions were as follows： pH 7.38, temperature 48.2~C and en- zyme/substrate （E/S）ratio 2 840 U/g. Under these conditions, the maximum DRSA was （22.85 ＋ 0.57）%. The experimental values agreed with the value （23.09%） predicted by the model within a 95% confidence interval. By using gel filtration chromatography and reversed-phase high performance liquid chromatography （RP-HPLC）, anti- oxidant peptide （D2-P）was isolated from flatfish skin protein hydrolysates （FSPH）and could exhibit a （54.28 ± 1.37） % scavenging activity on DPPH radical at the concentration of 5 mg/mL. This is the first report of a scientific basis for the preparation of antioxidant peptides from flatfish skin. The results suggested that the antioxidant pep- tides can be exploited into functional foods or used as a novel source of nutraceuticals.

Novel Antioxidant Peptides Derived from Enzymatic Hydrolysates of Macadamia Protein

In the present work, macadamia protein was enzymatic hydrolyzed to produce peptides which had abundant antioxidant activities. The relative antioxidant capacity was investigated through some in vitro models such as scavenging activity of DPPH radical, scavenging of ABTS+ radical and evaluation of the total antioxidant capacity assay. Macadamia protein was characterized by methods of DEAE cellulose cation-exchange chromatography and SDS-PAGE. Besides, method of price graph was used to compare the difference and to investigate the connection between the actual and ideal antioxidant value of the hydrolysates, aiming to reduce this difference.

Protective effects of peptide KSPLY derived from Hericium erinaceus on H_(2)O_(2)-induced oxidative damage in HepG2 cells

Reactive oxygen species(ROS)-induced oxidative damage is strongly associated with the pathogenesis of chronic diseases,and natural antioxidant peptides have good abilities of scavenging ROS.The antioxidant activity of peptide Lys-Ser-Pro-Leu-Tyr(KSPLY)derived from Hericium erinaceus remains unclear.In the present study,the antioxidant effect and mechanism of KSPLY on H_(2)O_(2)-induced oxidative damage in HepG2 cells were investigated.The results indicated that KSPLY exhibited the antioxidant capacity in H_(2)O_(2)-induced HepG2 cells by enhancing superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and catalase(CAT)activities.In comparison with the H_(2)O_(2)-treated damage group,the apoptosis rate,ROS level,and malondialdehyde(MDA)content of HepG2 cells treated with KSPLY were significantly decreased.The H.erinaceus-derived peptide KSPLY pretreatment promoted the expression of detoxification and antioxidant enzymes via the Keap1/Nrf2 signal pathway,thereby inhibiting the generation of ROS and MDA.In conclusion,the H.erinaceus-derived peptide KSPLY effectively protected HepG2 cells against H_(2)O_(2)-induced oxidative damage,and it provided a theoretical basis for the further development of new natural antioxidants.

Effects of brewers' spent grain protein hydrolysates on gas production, ruminal fermentation characteristics, microbial protein synthesis and microbial community in an artificial rumen fed a high grain diet

Background: Brewers' spent grain(BSG) typically contains 20% – 29% crude protein(CP) with high concentrations of glutamine, proline and hydrophobic and non-polar amino acid, making it an ideal material for producing valueadded products like bioactive peptides which have antioxidant properties. For this study, protein was extracted from BSG, hydrolyzed with 1% alcalase and flavourzyme, with the generated protein hydrolysates(AlcH and FlaH)showing antioxidant activities. This study evaluated the effects of AlcH and FlaH on gas production, ruminal fermentation characteristics, nutrient disappearance, microbial protein synthesis and microbial community using an artificial rumen system(RUSITEC) fed a high-grain diet.Results: As compared to the control of grain only, supplementation of FlaH decreased(P < 0.01) disappearances of dry matter(DM), organic matter(OM), CP and starch, without affecting fibre disappearances;while AlcH had no effect on nutrient disappearance. Neither AlcH nor FlaH affected gas production or VFA profiles, however they increased(P < 0.01) NH_3-N and decreased(P < 0.01) H_2 production. Supplementation of FlaH decreased(P < 0.01)the percentage of CH_4 in total gas and dissolved-CH_4(dCH_4) in dissolved gas. Addition of monensin reduced(P < 0.01) disappearance of nutrients, improved fermentation efficiency and reduced CH_4 and H_2 emissions.Total microbial nitrogen production was decreased(P < 0.05) but the proportion of feed particle associated(FPA) bacteria was increased with FlaH and monensin supplementation. Numbers of OTUs and Shannon diversity indices of FPA microbial community were unaffected by AlcH and FlaH;whereas both indices were reduced(P < 0.05) by monensin. Taxonomic analysis revealed no effect of AlcH and FlaH on the relative abundance(RA) of bacteria at phylum level, whereas monensin reduced(P < 0.05) the RA of Firmicutes and Bacteroidetes and enhanced Proteobacteria. Supplementation of FlaH enhanced(P < 0.05) the RA of genus Prevotella, reduced Selenomonas, Shuttleworthia, Bifidobacterium and Dialister as compared to control;monensin reduced(P < 0.05) RA of genus Prevotella but enhaced Succinivibrio.Conclusions: The supplementation of FlaH in high-grain diets may potentially protect CP and starch from ruminal degradation, without adversely affecting fibre degradation and VFA profiles. It also showed promising effects on reducing CH_4 production by suppressing H_2 production. Protein enzymatic hydrolysates from BSG using flavourzyme showed potential application to high value-added bio-products.

Endoplasmic reticulum-mitochondrial crosstalk:a novel role for the mitochondrial peptide humanin

In this review,the interactive mechanisms of mitochondria with the endoplasmic reticulum（ER） are discussed with emphasis on the potential protective role of the mitochondria derived peptide humanin（HN） in ER stress.The ER and mitochondria are dynamic organelles capable of modifying their structure and function in response to changing environmental conditions.The ER and mitochondria join together at multiple sites and form mitochondria-ER associated membranes that participate in signal transduction pathways that are under active investigation.Our laboratory previously showed that HN protects cells from oxidative stress induced cell death and more recently,described the beneficial role of HN on ER stress-induced apoptosis in retinal pigment epithelium cells and the involvement of ER-mitochondrial cross-talk in cellular protection.The protection was achieved,in part,by the restoration of mitochondrial glutathione that was depleted by ER stress.Thus,HN may be a promising candidate for therapy for diseases that involve both oxidative and ER stress.Developing novel approaches for retinal delivery of HN,its analogues as well as small molecular weight ER stress inhibitors would prove to be a valuable approach in the treatment of age-related macular degeneration.