Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 112
Filtrar
1.
Mar Drugs ; 17(5)2019 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-31035632

RESUMO

In this report, protein hydrolysate (TGH) of blood cockle (Tegillarca granosa) was prepared using a two-enzyme system (Alcalase treatment for 1.5 h following Neutrase treatment for 1.5 h). Subsequently, six antioxidant peptides were isolated from TGH using ultrafiltration and chromatography methods, and their amino acid sequences were identified as EPLSD, WLDPDG, MDLFTE, WPPD, EPVV, and CYIE with molecular weights of 559.55, 701.69, 754.81, 513.50, 442.48, and 526.57 Da, respectively. In which, MDLFTE and WPPD exhibited strong scavenging activities on DPPH radical (EC50 values of 0.53 ± 0.02 and 0.36 ± 0.02 mg/mL, respectively), hydroxy radical (EC50 values of 0.47 ± 0.03 and 0.38 ± 0.04 mg/mL, respectively), superoxide anion radical (EC50 values of 0.75 ± 0.04 and 0.46 ± 0.05 mg/mL, respectively), and ABTS cation radical (EC50 values of 0.96 ± 0.08 and 0.54 ± 0.03 mg/mL, respectively). Moreover, MDLFTE and WPPD showed high inhibiting ability on lipid peroxidation. However, MDLFTE and WPPD were unstable and could not retain strong antioxidant activity at high temperatures (>80 °C for 0.5 h), basic pH conditions (pH > 9 for 2.5 h), or during simulated GI digestion. In addition, the effect of simulated gastrointestinal digestion on TGP4 was significantly weaker than that on MDLFTE. Therefore, MDLFTE and WPPD may be more suitable for serving as nutraceutical candidates in isolated forms than as food ingredient candidates in functional foods and products.


Assuntos
Organismos Aquáticos , Bivalves , Depuradores de Radicais Livres/farmacologia , Peptídeos/farmacologia , Hidrolisados de Proteína/química , Sequência de Aminoácidos , Animais , Suplementos Nutricionais , Depuradores de Radicais Livres/química , Depuradores de Radicais Livres/isolamento & purificação , Alimento Funcional , Temperatura Alta , Concentração de Íons de Hidrogênio , Peroxidação de Lipídeos/efeitos dos fármacos , Peptídeos/química , Peptídeos/isolamento & purificação , Hidrolisados de Proteína/isolamento & purificação
2.
Int J Mol Sci ; 20(8)2019 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-31009989

RESUMO

The valorization of by-products from natural organic sources is an international priority to respond to environmental and economic challenges. In this context, electrodialysis with filtration membrane (EDFM), a green and ultra-selective process, was used to separate peptides from salmon frame protein hydrolysate. For the first time, the simultaneous separation of peptides by three ultrafiltration membranes of different molecular-weight exclusion limits (50, 20, and 5 kDa) stacked in an electrodialysis system, allowed for the generation of specific cationic and anionic fractions with different molecular weight profiles and bioactivity responses. Significant decreases in peptide recovery, yield, and molecular weight (MW) range were observed in the recovery compartments depending on whether peptides had to cross one, two, or three ultrafiltration membranes. Moreover, the Cationic Recovery Compartment 1 fraction demonstrated the highest increase (42%) in glucose uptake on L6 muscle cells. While, in the anionic configuration, both Anionic Recovery Compartment 2 and Anionic Recovery Compartment 3 fractions presented a glucose uptake response in basal condition similar to the insulin control. Furthermore, Cationic Recovery Compartment 3 was found to contain inhibitory peptides. Finally, LC-MS analyses of the bioassay-guided bioactive fractions allowed us to identify 11 peptides from salmon by-products that are potentially responsible for the glucose uptake improvement.


Assuntos
Glucose/metabolismo , Peptídeos/isolamento & purificação , Hidrolisados de Proteína/isolamento & purificação , Salmão/metabolismo , Animais , Ânions , Cátions , Linhagem Celular , Diálise , Camundongos , Peso Molecular , Termodinâmica , Ultrafiltração
3.
Mar Drugs ; 17(4)2019 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-30935056

RESUMO

A protein extract was generated from the macroalga Ulva lactuca, which was subsequently hydrolysed using the food-grade enzyme papain and angiotensin-converting Enzyme I and renin inhibitory peptides identified using a combination of enrichment strategies employing molecular weight cutoff filtration and mass spectrometry analysis. The generated hydrolysates with the most promising in vitro activity were further purified using preparative RP-HPLC and characterised. The 1 kDa hydrolysate (1 kDa-UFH), purified and collected by preparative RP-HPLC at minutes 41‒44 (Fr41‒44), displayed statistically higher ACE-I inhibitory activities ranging from 96.91% to 98.06%. A total of 48 novel peptides were identified from these four fractions by LC-MS/MS. A simulated gastrointestinal digestion of the identified peptide sequences was carried out using in silico enzyme cleavage simulation tools, resulting in 86 peptide sequences that were further assessed for their potential activity, toxicity and allergenicity using multiple predictive approaches. All the peptides obtained in this study were predicted to be non-toxic. However, 28 out of the 86 novel peptides released after the in silico gastrointestinal digestion were identified as potential allergens. The potential allergenicity of these peptides should be further explored to comply with the current labelling regulations in formulated food products containing U. lactuca protein hydrolysates.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Ulva/metabolismo , Alérgenos/farmacologia , Sequência de Aminoácidos , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Anti-Hipertensivos/farmacologia , Simulação por Computador , Humanos , Hidrólise , Oligopeptídeos/química , Oligopeptídeos/isolamento & purificação , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Alga Marinha/química , Ulva/química , Ulva/citologia
4.
Mar Drugs ; 17(3)2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30818811

RESUMO

The objective of this report was to investigate the isolation and recovery of different biocompounds and bioproducts from wastes (skins and heads) that were obtained from five species discarded by fishing fleets (megrim, hake, boarfish, grenadier, and Atlantic horse mackerel). Based on chemical treatments, enzymatic hydrolysis, and bacterial fermentation, we have isolated and produced gelatinous solutions, oils that are rich in omega-3, fish protein hydrolysates (FPHs) with antioxidant and antihypertensive activities, and peptones. FPHs showed degrees of hydrolysis higher than 13%, with soluble protein concentrations greater than 27 g/L and in vitro digestibilities superior to 90%. Additionally, amino acids compositions were always valuable and bioactivities were, in some cases, remarkable. Peptones that were obtained from FPHs of skin and the heads were demonstrated to be a viable alternative to expensive commercial ones indicated for the production of biomass, lactic acid, and pediocin SA-1 from Pediococcus acidilactici.


Assuntos
Produtos Biológicos/isolamento & purificação , Ácidos Graxos Ômega-3/isolamento & purificação , Peixes , Peptonas/isolamento & purificação , Hidrolisados de Proteína/isolamento & purificação , Animais , Anti-Hipertensivos/economia , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/farmacologia , Antioxidantes/economia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Bactérias/metabolismo , Produtos Biológicos/economia , Produtos Biológicos/farmacologia , Ácidos Graxos Ômega-3/economia , Ácidos Graxos Ômega-3/farmacologia , Fermentação , Pesqueiros/economia , Cabeça , Hidrólise , Peptonas/economia , Peptonas/farmacologia , Hidrolisados de Proteína/economia , Hidrolisados de Proteína/farmacologia , Pele/química , Espanha
5.
Mar Drugs ; 17(2)2019 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-30823522

RESUMO

Enhanced oxidative stress plays a central role in promoting endothelial dysfunction, leading to the development of atherosclerosis. In this study, we investigated the protective effects of the hydrolysates derived from blue mussel (Mytilus edulis) against H2O2-mediated oxidative injury in human umbilical vein endothelial cells (HUVECs). The blue mussel hydrolysates were prepared by enzymatic hydrolysis with eight proteases, and blue mussel-α-chymotrypsin hydrolysate (BMCH) showed the highest antioxidant activities in DPPH radical scavenging, ABTS⁺ radical scavenging, and ORAC value compared to those of the other hydrolysates. BMCH also inhibited Cu2+-mediated low density lipoprotein (LDL) oxidation. Treatment of H2O2 resulted in the decreased HUVEC viability whereas pre-treatment with BMCH increased HUVEC viability and reduced reactive oxygen species (ROS) generation. BMCH pre-treatment increased cellular antioxidant capacities, including levels of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) against H2O2-mediated oxidative stress in HUVECs. Flow cytometry and western blot analysis revealed that BMCH pre-treatment significantly reduced H2O2-mediated HUVEC apoptosis through inhibition of caspase-3 activation. Real-time-qPCR analysis showed that BMCH down-regulated expression of p53 and caspase-3 genes, as well as decreased the bax/bcl-2 ratio. Taken together, these results indicate that BMCH may be useful as functional food ingredients for protecting endothelial dysfunction or related disease.


Assuntos
Aminoácidos/química , Caspase 3/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Mytilus edulis/química , Estresse Oxidativo/efeitos dos fármacos , Hidrolisados de Proteína/farmacologia , Aminoácidos/metabolismo , Aminoácidos/farmacologia , Animais , Antioxidantes/química , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Glutationa/metabolismo , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Peróxido de Hidrogênio/administração & dosagem , Lipoproteínas LDL/metabolismo , Mytilus edulis/metabolismo , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro , Espécies Reativas de Oxigênio/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo
6.
Mar Drugs ; 17(3)2019 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-30893907

RESUMO

Angiotensin I-converting enzyme (ACE) inhibitory peptides derived from seaweed represent a potential source of new antihypertensive. The aim of this study was to isolate and purify ACE inhibitory peptides (ACEIPs) from the protein hydrolysate of the marine macroalga Ulva intestinalis. U. intestinalis protein was hydrolyzed by five different proteases (trypsin, pepsin, papain, α-chymotrypsin, alcalase) to prepare peptides; compared with other hydrolysates, the trypsin hydrolysates exhibited the highest ACE inhibitory activity. The hydrolysis conditions were further optimized by response surface methodology (RSM), and the optimum conditions were as follows: pH 8.4, temperature 28.5 °C, enzyme/protein ratio (E/S) 4.0%, substrate concentration 15 mg/mL, and enzymolysis time 5.0 h. After fractionation and purification by ultrafiltration, gel exclusion chromatography and reverse-phase high-performance liquid chromatography, two novel purified ACE inhibitors with IC50 values of 219.35 µM (0.183 mg/mL) and 236.85 µM (0.179 mg/mL) were obtained. The molecular mass and amino acid sequence of the ACE inhibitory peptides were identified as Phe-Gly-Met-Pro-Leu-Asp-Arg (FGMPLDR; MW 834.41 Da) and Met-Glu-Leu-Val-Leu-Arg (MELVLR; MW 759.43 Da) by ultra-performance liquid chromatography-tandem mass spectrometry. A molecular docking study revealed that the ACE inhibitory activities of the peptides were mainly attributable to the hydrogen bond and Zn(II) interactions between the peptides and ACE. The results of this study provide a theoretical basis for the high-valued application of U. intestinalis and the development of food-derived ACE inhibitory peptides.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Anti-Hipertensivos/farmacologia , Peptídeos/farmacologia , Peptidil Dipeptidase A/metabolismo , Ulva , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Anti-Hipertensivos/química , Anti-Hipertensivos/isolamento & purificação , Anti-Hipertensivos/uso terapêutico , Estabilidade de Medicamentos , Ensaios Enzimáticos , Hidrólise , Hipertensão/tratamento farmacológico , Concentração Inibidora 50 , Simulação de Acoplamento Molecular , Peptídeo Hidrolases/metabolismo , Peptídeos/química , Peptídeos/isolamento & purificação , Peptídeos/uso terapêutico , Peptidil Dipeptidase A/química , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Hidrolisados de Proteína/metabolismo , Alga Marinha/química
7.
Environ Sci Pollut Res Int ; 26(9): 8875-8884, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30715703

RESUMO

Fish protein hydrolysates are digested form of protein with various bioactive properties where, the cleavages of molecular bonds of proteins can be broken by the enzymatic and chemical process. In this study, antioxidant properties of spray dried protein hydrolysate prepared from Pangasius viscera by using enzymatic (papain and pepsin), and chemical methods (hydrochloric acid and sodium hydroxide) were evaluated. Among the different treatments, pepsin-derived visceral protein hydrolysate showed the maximum antioxidant activity when used at higher concentrations. Essential amino acids (EAA) and hydrophobic amino acids are higher in papain-derived visceral protein hydrolysate. In pepsin-derived visceral protein hydrolysate, major proportion was contributed by glycine (Gly), glutamine (Glu), proline (Pro), and asparagine (Asp). Higher amount of aromatic amino acids are found in alkali-derived FVPH. Scanning electron microscopy (SEM) images of pepsin fish visceral protein hydrolysate showed better globular structure than the other treatments. It can be concluded that among the different treatments, the visceral protein hydrolysate prepared with pepsin had better overall quality regarding antioxidant properties and papain in nutritional point of view.


Assuntos
Antioxidantes/farmacologia , Peixes-Gato , Hidrolisados de Proteína/farmacologia , Animais , Antioxidantes/química , Antioxidantes/isolamento & purificação , Papaína/química , Pepsina A/farmacologia , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Alimentos Marinhos/análise
8.
Poult Sci ; 98(6): 2360-2370, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30668770

RESUMO

Antioxidant peptides are increasingly attracting researchers in medicine and foods. In the present study, egg white hydrolysates of embryonated eggs hatched on the sixth day (EWHD) were segmented consecutively by ultrafiltration membranes with small tangential flow ultrafiltration system. Four segments with more than 30, 30 to 10 kDa, 10 to 5 kDa, and less than 5 kDa of molecular weight cut-off (MWCO) values were separated and were labeled as MWCOI, MWCOII, MWCOIII, and MWCOIV, respectively. The antioxidant activities of segments were investigated by performing DPPH•, •OH radical scavenging, ultra oxygen anion (O2-•), total antioxidant capacity, and reducing power experiments. The results indicated that MWCOI has the strongest scavenging activities on DPPH• radical. However, MWCOIV has the strongest scavenging activities on •OH, O2-•, total antioxidant capacity, and reducing power, which revealed that MWCOIV has strong antioxidant activity. MWCOIV was further separated into 14 fractions via semipreparative reverse-phased high-performance liquid chromatography (RP-HPLC), and their antioxidant activity was evaluated by different antioxidant assays in vitro. The fractions 10 and 7 had strong antioxidant activities. The purities of these 2 fractions were determined by analytical RP-HPLC. Moreover, both fractions 10 and 7 displayed high purity levels, and they were identified by quadrupole time-of-flight tandem mass spectrometry. Only fraction 10, with a molecular weight of 204 Da, can be identified to be Ser-Val. EWHD can be considered as a promising source of natural food antioxidants for the development of functional food.


Assuntos
Antioxidantes/isolamento & purificação , Proteínas Aviárias/isolamento & purificação , Embrião de Galinha/química , Peptídeos/isolamento & purificação , Animais , Antioxidantes/química , Proteínas Aviárias/química , Galinhas , Cromatografia Líquida de Alta Pressão/veterinária , Clara de Ovo/química , Óvulo/química , Peptídeos/química , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Espectrometria de Massas em Tandem/veterinária
9.
Food Chem ; 272: 453-461, 2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-30309568

RESUMO

This study follows recent attempts to discover natural xanthine oxidase (XO) inhibitors from foods, focusing herein on under-researched fish proteins. The anti-hyperuricemic function of tuna flesh hydrolysate (TPH) produced using Alcalase 2.4L was confirmed in potassium oxonate-induced hyperuricemic rats. TPH was separated using 80 wt% aqueous ethanol. The ethanol-soluble fraction (ESF) abundant in small peptides (<1000 Da) afforded the highest XO inhibition. Separation of ESF by Sephadex G-15 and UPLC/MS/MS revealed 13 di-/tri-peptides (12 are newly identified XO inhibitors). Their XO inhibitory activities were assessed using corresponding synthetic peptides via an improved HPLC method. Results indicate that Phe-containing peptides were more potent XO inhibitors than Trp-containing peptides, with Phe-His having the highest XO inhibitory activity (IC50 = 25.7 mM). Molecular docking studies revealed the importance of two hydrogen bonds and one π-π stacking interaction with Phe-914 in XO for XO-peptide inhibitor binding. Phe-containing di-/tri-peptides could be potent XO inhibitors against hyperuricemia.


Assuntos
Inibidores Enzimáticos/uso terapêutico , Hiperuricemia/tratamento farmacológico , Hidrolisados de Proteína/metabolismo , Atum/metabolismo , Xantina Oxidase/antagonistas & inibidores , Animais , Sítios de Ligação , Cromatografia Líquida de Alta Pressão , Inibidores Enzimáticos/metabolismo , Etanol/química , Ligações de Hidrogênio , Hiperuricemia/induzido quimicamente , Hiperuricemia/veterinária , Simulação de Acoplamento Molecular , Ácido Oxônico/toxicidade , Peptídeos/análise , Peptídeos/uso terapêutico , Hidrolisados de Proteína/isolamento & purificação , Estrutura Terciária de Proteína , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem , Xantina Oxidase/metabolismo
10.
Molecules ; 23(7)2018 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-30029493

RESUMO

The high-pressure homogenization (HPH) treatment of soybean protein isolate (SPI) before enzymatic hydrolysis using bromelain was investigated. Homogenization pressure and cycle effects were evaluated on the enzymatic degree of hydrolysis and the antioxidant activity of the hydrolysates generated. The antioxidant activity of SPI hydrolysates was analyzed by 1,1-dipheny-2-picrylhydrazyl (DPPH). The sizes and structures of the SPI-soluble aggregate after HPH treatment were analyzed using dynamic and static laser light scattering. The changes in the secondary structure, as measured by Fourier transform infrared spectroscopy (FTIR) and the macromorphology of SPI, were measured by scanning electron microscope (SEM). These results suggested that the HPH treatment (66.65%) could increase the antioxidant activities of the SPI hydrolysates compared with the control (54.18%). SPI hydrolysates treated at 20 MPa for four cycles obtained higher DPPH radical-scavenging activity than other samples. The control was predicted to be a hard sphere, and SPI treatment at 10 MPa was speculated to be Gaussian coil, polydisperse, and then the high-pressure treated SPI became a hollow sphere. Changes in the secondary structures showed protein aggregate formation and rearrangements. The image of SPI varied from a globular to a clump structure, as observed by the SEM. In conclusion, combining HPH treatment and enzymolysis could be an effective way to improve the antioxidant activity of the SPI.


Assuntos
Pressão , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Proteínas de Soja/química , Proteínas de Soja/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Catálise , Difusão Dinâmica da Luz , Hidrólise , Modelos Moleculares , Agregados Proteicos , Conformação Proteica , Hidrolisados de Proteína/ultraestrutura , Proteínas de Soja/ultraestrutura , Espectroscopia de Infravermelho com Transformada de Fourier
11.
Mar Drugs ; 16(7)2018 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-29997311

RESUMO

Hydrolysates of food protein sources have immunomodulatory effects, which are of interest for use as functional foods. In this study, we have characterized the immune regulatory effect on rat splenocytes, macrophages and T lymphocytes of Ulva spp. hydrolysates and their peptide fractions with or without in vitro gastrointestinal digestion and/or ultrafiltration. IL-10 was induced in almost all conditions and cell types obtained from wild type animals. The induction was in general increased by ultrafiltration and in vitro gastrointestinal digestion. TNF was also induced in basal conditions. In turn, TNF and IFN-γ production was attenuated by the hydrolysate products in lipopolysaccharide or concanavalin A immune stimulated cells. Inhibitors for the activation of NFκB, MAPK p38 and JNK inhibited IL-10 induction in rat splenocytes. The response was dramatically attenuated in TLR4-/- cells, and only modestly in TLR2-/- cells. Food peptides from Ulva spp. genus exert anti-inflammatory effects in immune cells mediated by TLR4 and NFκB. Similarity with the immunomodulatory profile of protein hydrolysates from other sources suggests a common mechanism.


Assuntos
Citocinas/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Peptídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Ulva/química , Animais , Células Cultivadas , Feminino , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Peptídeos/isolamento & purificação , Cultura Primária de Células , Hidrolisados de Proteína/isolamento & purificação , Ratos , Ratos Wistar , Baço/citologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
12.
Food Chem ; 261: 301-310, 2018 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-29739598

RESUMO

In this study, the hydrolysis of a pecan protein isolate (PPI) with Alcalase was carried out to generate antioxidant peptides. We proposed a kinetic model to illustrate the enzymolysis process of PPI, which was found suitable for depiction of the kinetic behavior for PPI hydrolysis by Alcalase. The PPI hydrolysis products were gradually fractionated by ultrafiltration through cut-off membranes with molecular weights of 10, 5 and 3 kDa and their antioxidant activities were evaluated in vitro. Further, the strongest antioxidant fraction (<3 kDa) and novel antioxidative peptide were successfully purified. The amino acid sequence of the purified peptide was identified as LAYLQYTDFETR. The purified fraction exhibited appreciable scavenging activities on ABTS radical (67.67%), DPPH radical (56.25%) and hydroxyl radical (47.42%) at 0.1 mg/mL. The results suggested that this novel peptide may serve as a potential antioxidant and it should be evaluated for development of functional foods and pharmaceuticals products.


Assuntos
Antioxidantes/química , Carya/química , Peptídeos/química , Subtilisinas/química , Sequência de Aminoácidos , Antioxidantes/isolamento & purificação , Biocatálise , Hidrólise , Cinética , Peso Molecular , Peptídeos/isolamento & purificação , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Ultrafiltração
13.
Plant Foods Hum Nutr ; 73(2): 101-107, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29679358

RESUMO

In previous studies, it has not been reported that protein isolated from chia interferes favorably with antibacterial activity, and reduces cholesterol synthesis. The objective of this study was to determine whether commonly used commercial microbial proteases can be utilized to generate chia protein-based antibacterial and hypocholesterolemic hydrolysates/peptides, considering the effects of protein extraction method. Alcalase, Flavourzyme and sequential Alcalase-Flavourzyme were used to produce hydrolysates from chia protein (CF), protein-rich fraction (PRF) and chia protein concentrates (CPC1 and CPC2). These hydrolysates were evaluated for their antimicrobial activity against Gram-positive (G+) and Gram-negative (G-) microorganisms. The protein hydrolysates were purified by ultrafiltration through a membrane with 3 kDa nominal molecular weight, for evaluation of hypocholesterolemic activity. An inhibition zone was observed when the hydrolysate was tested against S. aureus, and minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values were obtained. Peptides from chia protein with molecular mass lower than 3 kDa reduced up to 80.7% of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) enzymatic reaction velocity. It was also observed that, independent of the method used to obtain chia proteins, the fractions showed relevant bioactivity. Moreover, the intensity of the bioactivity varied with the method for obtaining the protein and with the enzyme used in the hydrolysis process. This is the first report to demonstrate that chia peptides are able to inhibit cholesterol homeostasis.


Assuntos
Antibacterianos/farmacologia , Anticolesterolemiantes/farmacologia , Peptídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Salvia/química , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/análise , Antibacterianos/isolamento & purificação , Anticolesterolemiantes/análise , Anticolesterolemiantes/isolamento & purificação , Colesterol , Endopeptidases/metabolismo , Hidrólise , Peso Molecular , Peptídeo Hidrolases/metabolismo , Peptídeos/análise , Peptídeos/isolamento & purificação , Hidrolisados de Proteína/análise , Hidrolisados de Proteína/isolamento & purificação , Subtilisinas/metabolismo
14.
Food Chem ; 254: 36-46, 2018 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-29548465

RESUMO

Two samples of trout frame protein hydrolysates were prepared by Microwave Pretreatment followed by Conventional Enzymatic hydrolysis (MPCE) and Non-Pretreated followed by Microwave-assisted Enzymatic hydrolysis (NPME), respectively, were subjected to simulated gastrointestinal digestion. Changes on degree of hydrolysis, antioxidant activity, molecular weight, and amino acid composition between undigested and after gastrointestinal digestion of peptides were investigated. Comparing to undigested peptides, a breakdown of MPCE and NPME into smaller molecules was observed. Degree of hydrolysis, ABTS+ radical scavenging activity and reducing power increased (P < 0.05) for both samples after gastrointestinal digestion. A purified peptide from GI-MPCE had two possible sequences, NGRLGYSEGVM or GNRLGYSWDD (1182.65 Da). Whereas GI-NPME had two peptides IRGPEEHMHR or RVAPEEHMHR (1261.77 Da) and SAGVPRHK or SARPRHK (962.63 Da). These results indicate that digested hydrolysates can be a rich source of antioxidants. Isolated peptides extracted from trout frame by-products could be new food ingredients used as natural antioxidants.


Assuntos
Antioxidantes/análise , Antioxidantes/isolamento & purificação , Biomimética , Digestão , Micro-Ondas , Hidrolisados de Proteína/análise , Hidrolisados de Proteína/isolamento & purificação , Sequência de Aminoácidos , Antioxidantes/química , Trato Gastrointestinal/fisiologia , Hidrólise , Peso Molecular , Hidrolisados de Proteína/química
15.
Food Chem ; 253: 101-107, 2018 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29502808

RESUMO

Spent hens have low market value and incur significant costs for disposal. This study explored the use of spent hens as a source of bioactive peptides. Spent hen hydrolysates prepared by Protease M or Protex 50FP exhibited interleukin (IL)-6 inhibitory activity in endotoxin-activated macrophage-like U937 cells (p < .05). The potential peptides from Protex 50FP hydrolysate were further fractionated using a combination of ultrafiltration, solid-phase extraction and high-performance liquid chromatography; 17 novel peptides encoded in major muscle proteins were identified by mass spectrometry analysis, of which 7 were chemically synthesized and assayed for the IL-6 inhibitory activity. At a concentration of 100 µg/mL, peptide FLWGKSY induced a 79% reduction of IL-6 production in endotoxin-activated macrophage-like U937 cells, which is comparable to results reported from other food sources. Our results indicate that spent hens have potential to be a source of bioactive peptides for anti-inflammatory applications.


Assuntos
Anti-Inflamatórios/isolamento & purificação , Galinhas/metabolismo , Interleucina-6/antagonistas & inibidores , Proteínas Musculares/química , Peptídeos/isolamento & purificação , Hidrolisados de Proteína/química , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Cromatografia Líquida de Alta Pressão/veterinária , Feminino , Humanos , Espectrometria de Massas/veterinária , Proteínas Musculares/isolamento & purificação , Peptídeos/química , Peptídeos/farmacologia , Hidrolisados de Proteína/isolamento & purificação , Extração em Fase Sólida/veterinária , Células U937
16.
Mol Med Rep ; 17(4): 5306-5311, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29393343

RESUMO

Liposomal angiotensin-I-converting enzyme inhibitory (ACEI) peptides were prepared from sunflower protein hydrolysates by the thin­film ultrasonic method. Response surface methodology (RSM), in combination with fractional factorial designs and central composite design methods were utilized to optimize entrapment efficiency and balance the drug release. We found that the ratio of phospholipids to cholesterol, ultrasound time and the ratio of phospholipids to ACEI peptides were significant factors affecting entrapment efficiency (P<0.001). Optimal preparation conditions of liposomal­ACEI peptides were the ratio of soybean phospholipids to cholesterol (w/w) of 4.1:1, PEG­2000 dosage (%) of 4, NaCl concentration in PBS (mM) of 50, hydration temperature of 45˚C, ultrasound time of 8.05 min and the ratio of soybean phospholipids to ACEI peptides of 15:1 (w/w). The experimental entrapment efficiency of liposomal­ACEI peptides was (91.25±0.182%). Moreover, the balanced release rate of liposome encapsulated ACEI in phosphate buffer was 77.83% after 12 h.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Helianthus/química , Lipossomos , Peptídeos/administração & dosagem , Hidrolisados de Proteína/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Helianthus/metabolismo , Lipossomos/química , Peptídeos/química , Peptídeos/isolamento & purificação , Peptidil Dipeptidase A/metabolismo , Fosfolipídeos/química , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Hidrolisados de Proteína/química , Hidrolisados de Proteína/isolamento & purificação , Reprodutibilidade dos Testes
17.
Molecules ; 23(2)2018 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-29393865

RESUMO

In recent years, food proteins with bioactivity have been studied for cancer treatment. Zein peptides have shown an important set of bioactivities. This work compares the cytotoxic activity of zein hydrolyzed, extracted from four Zea species: teosinte, native, hybrid, and transgenic (Teo, Nat, Hyb, and HT) in a hepatic cell culture. Zein fraction was extracted, quantified, and hydrolyzed. Antioxidant capacity and cytotoxicity assays were performed on HepG2 cells. The levels of expression of caspase 3, 8, and 9 were evaluated in zein-treated cell cultures. Zea parviglumis showed the highest zein content (46.0 mg/g) and antioxidant activity (673.40 TE/g) out of all native zeins. Peptides from Hyb and HT showed high antioxidant activity compared to their native counterparts (1055.45 and 724.32 TE/g, respectively). Cytotoxic activity was observed in the cell culture using peptides of the four Zea species; Teo and Nat (IC50: 1781.63 and 1546.23 ng/mL) had no significant difference between them but showed more cytotoxic activity than Hyb and HT (IC50: 1252.25 and 1155.56 ng/mL). Increased expression of caspase 3 was observed in the peptide-treated HepG2 cells (at least two-fold more with respect to the control sample). These data indicate the potential for zein peptides to prevent or treat cancer, possibly by apoptosis induction.


Assuntos
Antioxidantes/farmacologia , Citotoxinas/farmacologia , Regulação da Expressão Gênica de Plantas , Hidrolisados de Proteína/farmacologia , Zeína/farmacologia , Antioxidantes/isolamento & purificação , Caspase 3/genética , Caspase 3/metabolismo , Caspase 8/genética , Caspase 8/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Citotoxinas/isolamento & purificação , Células Hep G2 , Humanos , Concentração Inibidora 50 , Plantas Geneticamente Modificadas , Hidrolisados de Proteína/isolamento & purificação , Especificidade da Espécie , Zea mays/química , Zea mays/genética , Zea mays/metabolismo , Zeína/isolamento & purificação
18.
Molecules ; 24(1)2018 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-30597854

RESUMO

The protein by-products from carp (Cyprinus carpio) are normally discarded as industrial waste during fish processing. The objective of this study was to identify and characterise the peptides with a potential antioxidant activity that are released from carp skin proteins during hydrolysis by the Protamex enzyme mixture. This study shows that a hydrolysate of carp skin gelatin and its reversed-phase chromatography fractions have strong in vitro antioxidant properties. Among these fractions, the alanine-tyrosine (Ala-Tyr) dipeptide was identified as the major compound with high antioxidant potential. The peptide has good stability during in vitro enzymatic digestion assay and can inhibit the angiotensin-converting enzyme (ACE). In conclusion, our study proves that both the unfractionated hydrolysate of carp skin gelatin and the above-mentioned Ala-Tyr dipeptide represents attractive novel compounds for the formulation of antioxidant foods.


Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Carpas/metabolismo , Gelatina/química , Peptídeos/química , Peptídeos/farmacologia , Hidrolisados de Proteína/química , Hidrolisados de Proteína/farmacologia , Pele/química , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Antioxidantes/síntese química , Antioxidantes/isolamento & purificação , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Gelatina/metabolismo , Hidrólise , Espectrometria de Massas , Peptídeos/síntese química , Peptídeos/isolamento & purificação , Hidrolisados de Proteína/isolamento & purificação , Pele/metabolismo
19.
Nutrients ; 9(9)2017 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-28878183

RESUMO

Gastritis or peptic ulcer is believed to affect about half of people worldwide. Traditional medications can lead to adverse effects, therefore, alternative nutritional strategies are needed to prevent the development of gastric mucosal damage. A novel combination of two food-grade ingredients, wheat peptides and fucoidan (WPF), was prepared to treat male Sprague Dawley rats for 30 days before gastric mucosal damage was induced by oral administration of ethanol. The serum levels of biomarkers were determined by enzyme-linked immunosorbent assay. Biomarkers in stomach tissue were analyzed using immunohistochemistry. In addition, human gastric epithelial cell line (GES-1) was used to investigate protein expression by Western blot. WPF could attenuate ethanol-induced gastric mucosal damage in an inverse dose-dependent manner, with both ulcer index and pathological index improved. WPF increased superoxide dismutase level and decreased malondialdehyde level. WPF also decreased the levels of interleukin-8, platelet-activating factor, and Caspase 3, while increasing the levels of prostaglandin E-2, epidermal growth factor (EGF), and EGF receptor (EGFR). Furthermore, phosphorylation of EGFR and extracellular signal-regulated kinases was induced by WPF in GES-1 cells. In conclusion, the novel combination of wheat peptides and fucoidan attenuated ethanol-induced gastric mucosal damage in rats through anti-oxidant, anti-inflammatory, and pro-survival mechanisms.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Etanol , Mucosa Gástrica/efeitos dos fármacos , Proteínas de Plantas/farmacologia , Polissacarídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Úlcera Gástrica/prevenção & controle , Animais , Anti-Inflamatórios/isolamento & purificação , Antioxidantes/isolamento & purificação , Biomarcadores/sangue , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Receptores ErbB/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Humanos , Masculino , Fosforilação , Proteínas de Plantas/isolamento & purificação , Hidrolisados de Proteína/isolamento & purificação , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Úlcera Gástrica/sangue , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/patologia , Fatores de Tempo , Triticum/química
20.
Biol Pharm Bull ; 40(7): 984-991, 2017 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-28381791

RESUMO

Glutathione (GSH) is an ubiquitous thiol-containing tripeptide, which plays important roles in cellular protection from oxidative stress. In our search for a dietary source that can increase GSH levels, we discovered that a 24 h treatment of HepG2 cells with rice bran protein hydrolysate (RBPH), prepared by Umamizyme G-catalyzed hydrolysis, increased the GSH content in a dose-dependent manner. RBPH elevated the expression levels of γ-glutamylcysteine synthetase (γ-GCS), which constitutes the rate-limiting enzyme of GSH synthesis, and of another two enzymes, hemeoxygenase-1 (HO-1) and reduced nicotinamide adenine dinucleotide (phosphate): quinone oxidoreductase 1 (NQO1). This induction was preceded by the accumulation of nuclear factor erythroid 2-related factor 2 (Nrf2) inside the nucleus, which is a key transcription factor for the expression of the γ-GCS, HO-1, and NQO1. Pre-treatment of cells with RBPH produced a significant protective effect against cytotoxicity caused by H2O2 or ethanol. These results indicate that RBPH exerts a protective effect against oxidative stress by modulating GSH levels and anti-oxidative enzyme expression via the Nrf2 pathway.


Assuntos
Antioxidantes/farmacologia , Glutationa/metabolismo , Oryza/química , Estresse Oxidativo/efeitos dos fármacos , Proteínas de Plantas/química , Hidrolisados de Proteína/farmacologia , Antioxidantes/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Hep G2 , Humanos , Fator 2 Relacionado a NF-E2/metabolismo , Hidrolisados de Proteína/isolamento & purificação , Transdução de Sinais
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA