Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 17.847
Filtrar
1.
Methods Mol Biol ; 2259: 167-179, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33687714

RESUMO

Metaproteomics of host-microbiome interfaces comprises the analysis of complex mixtures of bacteria, archaea, fungi, and viruses in combination with its host cells. Microbial niches can be found all over the host including the skin, oral cavity, and the intestine and are considered to be essential for the homeostasis. The complex interactions between the host and diverse commensal microbiota are poorly characterized while of great interest as dysbiosis is associated with the development of various inflammatory and metabolic diseases. The metaproteomics workflows to study these interfaces are currently being established, and many challenges remain. The major challenge is the large diversity in species composition that make up the microbiota, which results in complex samples that require extended mass spectrometry analysis time. In addition, current database search strategies are not developed to the size of the search space required for unbiased microbial protein identification.Here, we describe a workflow for the proteomics analysis of microbial niches with a focus on intestinal mucus layer. We will cover step-by-step the sample collection, sample preparation, liquid chromatography-mass spectrometry, and data analysis.


Assuntos
Bactérias/isolamento & purificação , Proteínas de Bactérias/análise , Proteínas Fúngicas/análise , Fungos/isolamento & purificação , Microbioma Gastrointestinal , Proteômica/métodos , Animais , Cromatografia Líquida/métodos , Intestinos/microbiologia , Espectrometria de Massas/métodos , Camundongos , Peptídeos/análise , Fluxo de Trabalho
2.
Methods Mol Biol ; 2259: 215-223, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33687718

RESUMO

A workflow for the characterization of food-derived bioactive peptides is described in this chapter. The workflow integrates two consecutive steps: a discovery phase and a protein-based bioinformatic phase. In the first step (discovery phase), a shotgun bottom-up proteomics approach is used to create a reference data set for a selected food proteome. Afterward, in a second step (bioinformatic phase), the reference proteome is subjected to several in silico protein-based bioinformatic analyses to predict and characterize potential bioactive peptides after an in silico human gastrointestinal digestion. Using this workflow, bioactive collagen peptides, antihypertensive, antimicrobial, and antitumor peptides were predicted as potential valuable bioactive peptides from seafood and marine by-products. It is concluded that the combination of the global shotgun proteomic analysis and the analysis by protein-based bioinformatics can provide a rapid strategy for the characterization of new potential food-derived bioactive peptides.


Assuntos
Proteínas na Dieta/análise , Peptídeos/análise , Proteômica/métodos , Cromatografia Líquida/métodos , Alimento Funcional/análise , Humanos , Alimentos Marinhos/análise , Espectrometria de Massas em Tandem/métodos
3.
Food Chem ; 349: 129133, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33561795

RESUMO

The strategy of taste-guided assisted by solvent extraction, solid-phase extraction and semipreparative HPLC were applied to isolate the main nonvolatile bitter components from mechanized Huangjiu. The potential fraction was identified by amino acid analysis and ultra-performance liquid chromatography-quadrupole-time-of-flight-MS/MS. Bitter pyroglutamate peptide Pyr-LFNPSTNPWHSP (PGP) was successfully identified from Huangjiu for the first time. Quantitative analysis showed that PGP contents ranged from below the limit of quantitation to 32.97 mg/L, among mechanized Huangjiu had higher contents than manual and commercial Huangjiu. The formation of PGP mainly occurred in the primary fermentation and it was stable in Huangjiu. Moreover, the PGP content of the Huangjiu brewed using raw wheat Qu was 112.6% higher than that using cooked wheat Qu, but presented subtle change with the increase of raw wheat Qu. The results revealed that PGP contributed the bitterness to Huangjiu, which may offer a possibility to reduce the bitterness of Huangjiu.


Assuntos
Bebidas Alcoólicas/análise , Análise de Alimentos , Paladar , Sequência de Aminoácidos , Fermentação , Humanos , Peptídeos/análise , Peptídeos/química , Peptídeos/isolamento & purificação
4.
Food Chem ; 345: 128810, 2021 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-33601654

RESUMO

The inability to easily identify the animal species in highly processed meat products makes them highly susceptible to adulterations. Reliable methods for detecting the species origin of meat used in processed food are required to ensure adequate labelling and minimize food fraud and allergenic potential. Liquid chromatography high resolution mass spectrometry was employed to identify new heat-stable guinea-fowl-specific peptide markers that can differentiate guinea fowl meat from other commonly consumed animal species, including closely related poultry species, in highly processed food products. We identified 26 unique guinea-fowl-specific markers. The high stability of guinea-fowl-specific peptides was confirmed by analysing food products with guinea fowl meat content ranging from 4% to 100%. The findings indicate that sensitive and reliable LC-MS/MS methods can be developed for the targeted detection and quantification of guinea fowl meat in highly processed meat products.


Assuntos
Análise de Alimentos/métodos , Carne/análise , Peptídeos/análise , Sequência de Aminoácidos , Animais , Biomarcadores/análise , Cromatografia Líquida de Alta Pressão , Guiné , Peptídeos/química , Análise de Componente Principal , Espectrometria de Massas em Tandem
5.
J Chromatogr A ; 1639: 461922, 2021 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-33540183

RESUMO

A peak-tracking algorithm was developed for use in comprehensive two-dimensional liquid chromatography coupled to mass spectrometry. Chromatographic peaks were tracked across two different chromatograms, utilizing the available spectral information, the statistical moments of the peaks and the relative retention times in both dimensions. The algorithm consists of three branches. In the pre-processing branch, system peaks are removed based on mass spectra compared to low intensity regions and search windows are applied, relative to the retention times in each dimension, to reduce the required computational power by elimination unlikely pairs. In the comparison branch, similarity between the spectral information and statistical moments of peaks within the search windows is calculated. Lastly, in the evaluation branch extracted-ion-current chromatograms are utilized to assess the validity of the pairing results. The algorithm was applied to peptide retention data recorded under varying chromatographic conditions for use in retention modelling as part of method optimization tools. Moreover, the algorithm was applied to complex peptide mixtures obtained from enzymatic digestion of monoclonal antibodies. The algorithm yielded no false positives. However, due to limitations in the peak-detection algorithm, cross-pairing within the same peaks occurred and six trace compounds remained falsely unpaired.


Assuntos
Algoritmos , Anticorpos Monoclonais/análise , Cromatografia Líquida/métodos , Peptídeos/análise , Espectrometria de Massas/métodos , Reconhecimento Automatizado de Padrão , Padrões de Referência
6.
Zoolog Sci ; 38(1): 51-59, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33639718

RESUMO

In vertebrates, gonadotropin-releasing hormone (GnRH) regulates gonadal maturation by stimulating the synthesis and release of pituitary gonadotropins. GnRH has also been identified in invertebrates. Crustacea consists of several classes including Cephalocarida, Remipedia, Branchiopoda (e.g., tadpole shrimp), Hexanauplia (e.g., barnacle) and Malacostraca (e.g., shrimp, crab). In the malacostracan crustaceans, the presence of GnRH has been detected in several species, mainly by immunohistochemistry. In the present study, we examined whether a GnRH-like peptide exists in the brain and/or nerve ganglion of three classes of crustaceans, the tadpole shrimp Triops longicaudatus (Branchiopoda), the barnacle Balanus crenatus (Hexanauplia), and the hermit crab Pagurus filholi (Malacostraca), by immunohistochemistry using a rabbit polyclonal antibody raised against chicken GnRH-II (GnRH2). This antibody was found to recognize the giant freshwater prawn Macrobrachium rosenbergii GnRH (MroGnRH). In the tadpole shrimp, GnRH-like-immunoreactive (ir) cell bodies were located in the circumesophageal connective of the deuterocerebrum, and GnRH-like-ir fibers were detected also in the ventral nerve cord. In the barnacle, GnRH-like-ir cell bodies and fibers were located in the supraesophageal ganglion (brain), the subesophageal ganglion, and the circumesophageal connective. In the hermit crab, GnRH-like-ir cell bodies were detected in the anterior-most part of the supraesophageal ganglion and the subesophageal ganglion. GnRH-like-ir fibers were observed also in the thoracic ganglion and the eyestalk. These results suggest that a GnRH-like peptide exists widely in crustacean species.


Assuntos
Crustáceos/anatomia & histologia , Crustáceos/metabolismo , Gânglios/metabolismo , Hormônio Liberador de Gonadotropina/metabolismo , Animais , Imuno-Histoquímica , Peptídeos/análise
7.
J Chromatogr A ; 1639: 461877, 2021 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-33545578

RESUMO

An analytical approach using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed to simultaneously detect Fagopyrum esculentum Moench (buckwheat) and cereals containing gluten (Triticum species including wheat and spelt, rye, barley, and oats) that were specified in regulations for food allergen labeling on processed foods. Trypsin-digested peptides were purified from different processed food commodities and heptapeptides derived from buckwheat 13S globulin (GFIVQAR, m/z 395.8 [precursor] > 177.0 [product]) and Triticum low molecular weight glutenin (QIPEQSR, m/z 429.3 [precursor] > 616.2 [product]) were specifically detected each species at levels as low as 0.050-0.056 µg/L and 0.028-0.032 µg/L, respectively. Detection of these synthetic peptides was quantitative to over 100 µg/L by reference to the synthetic peptide calibration curves and at recovery rates, 76.6 ± 4.1%-104.8 ± 17.1% and 82.4 ± 2.0%-105.8 ± 5.3%, for GFIVQAR and QIPEQSR, respectively, when 1-1,000 µg of these peptides were spiked into a retort tomato sauce for pasta or dried instant soup. In combination with LC-MS/MS detection methods specific to other cereals containing gluten (rye, barley, and oats), the developed analytical approach was applicable to a wide variety of processed food commodities for food allergen labeling.


Assuntos
Alérgenos/análise , Grão Comestível/química , Análise de Alimentos , Hipersensibilidade Alimentar/diagnóstico , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão , Peptídeos/análise
8.
Food Chem ; 347: 129008, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33484958

RESUMO

Probiotics can release many bioactive peptides that confer a myriad of benefits to the host health. However, exploring new bioactive peptides secreted by probiotics is hampered by lots of matrix-related interference peptides from the medium, and the low abundance. To this end, a new approach integrating mixed-mode cationic exchange based solid-phase extraction (MCX-SPE) coupled with liquid chromatography-mass spectrometry (LC-MS) and feature-based molecular networking (FBMN) was developed. FBMN's intuitive visualization results enabled twenty-five novel peptides to be quickly discovered and characterized from the cultures of three strains of Bifidobacterium, B. animalis, B. longum, and B. pseudolongum. Interestingly, four were uniquely secreted by B. animalis treated with gypenosides, and one showed ACE inhibitory effect with an IC50 value of 193.22 µM. Consequently, this approach could serve as a powerful tool for quickly discovering bioactive peptides from the complex metabolites of probiotics, which contributes to the development of functional foods and nutraceuticals.


Assuntos
Bifidobacterium/metabolismo , Cromatografia Líquida/métodos , Espectrometria de Massas/métodos , Peptídeos/análise , Extração em Fase Sólida/métodos , Peptídeos/isolamento & purificação , Probióticos
9.
Food Chem ; 348: 129134, 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-33516993

RESUMO

In this study, similarities and differences of sodium tripolyphosphate (STPP) and sodium trimetaphosphate (STMP) pre-soaking on the stability of muscle proteins in shrimp were investigated during 12 weeks of frozen storage (-30 °C). The physicochemical analysis indicated significant improvements in the WHC, springiness, chewiness, and thermal stability of STPP and STMP pre-soaked samples when compared to the control. Interestingly, STMP pre-soaking showed better cryoprotective effects than the STPP treatment when the storage period reached the end of the 12 weeks. Furthermore, the label-free based proteomics results indicated that 62 upregulated differentially abundant proteins (DAPs) were detected in STMP when compared to STPP. These identified DAPs specifically included 40S ribosomal proteins, actin-related proteins, heat shock proteins, myosin heavy chain, and tubulin beta chain. Additionally, the gene ontology (GO) and eukaryotic clusters of orthologous group (KOG) analyses verified that the incorporation of STMP molecules enhanced the resistance of cytoskeleton proteins to cold-temperature stress.


Assuntos
Armazenamento de Alimentos/métodos , Penaeidae/metabolismo , Polifosfatos/química , Alimentos Marinhos/análise , Animais , Cromatografia Líquida de Alta Pressão , Congelamento , Proteínas de Choque Térmico/metabolismo , Penaeidae/química , Peptídeos/análise , Proteínas de Frutos do Mar/metabolismo , Espectrometria de Massas em Tandem , Fatores de Tempo
10.
Food Chem ; 348: 129097, 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-33515941

RESUMO

The aim of this study was to isolate and identify antibacterial peptides (ABPs) produced by lactic acid bacteria (LAB) in Chinese pickles. The cell-free supernatant collected from the culture of LAB with antibacterial activity against Staphylococcus aureus was used to purify ABPs. A total of 14 strains of LAB were found to have antibacterial activity. Among them, Lactobacillus fermentum (L. fermentum) SHY10 exhibited the most effective antibacterial activity. The antibacterial activity of cell-free supernatant reached the highest level after 20 h of L. fermentum SHY10 culture. Three novel ABPs were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In particular, the NQGPLGNAHR peptide showed antibacterial activity with an IC50 value of 0.957 mg/mL. In addition, molecular docking analysis revealed that this peptide interacted with DNA gyrase and dihydrofolate reductase by salt bridge formation, hydrogen bond interactions, and metal contact.


Assuntos
Antibacterianos/análise , Antibacterianos/isolamento & purificação , Lactobacillus fermentum/metabolismo , Peptídeos/análise , Peptídeos/isolamento & purificação , Antibacterianos/biossíntese , Alimentos e Bebidas Fermentados/microbiologia , Peptídeos/metabolismo
11.
Food Chem ; 340: 128154, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33010641

RESUMO

Numerous bacteria are responsible for hydrolysis of proteins during cheese ripening. The raw milk flora is a major source of bacterial variety, starter cultures are needed for successful acidification of the cheese and proteolytic strains like Lactobacillus helveticus, are added for flavor improvement or acceleration of ripening processes. To study the impact of higher bacterial diversity in cheese on protein hydrolysis during simulated human digestion, Raclette-type cheeses were produced from raw or heat treated milk, with or without proteolytic L. helveticus and ripened for 120 days. Kinetic processes were studied with a dynamic (DIDGI®) in vitro protocol and endpoints with the static INFOGEST in vitro digestion protocol, allowing a comparison of the two in vitro protocols at the level of gastric and intestinal endpoints. Both digestion protocols resulted in comparable peptide patterns after intestinal digestion and higher microbial diversity in cheeses led to a more diverse peptidome after simulated digestion.


Assuntos
Queijo/microbiologia , Proteínas do Leite/metabolismo , Leite/microbiologia , Aminoácidos/análise , Animais , Queijo/análise , Cromatografia Líquida de Alta Pressão , Digestão , Microbiologia de Alimentos , Humanos , Lactobacillus helveticus/genética , Lactobacillus helveticus/crescimento & desenvolvimento , Lactobacillus helveticus/metabolismo , Espectrometria de Massas , Leite/metabolismo , Peptídeos/análise , Proteólise , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo
12.
Food Chem ; 340: 128152, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33032150

RESUMO

Soy protein isolates were fermented by three commercial Lactobacillus helveticus strains for a maximum of seven days to investigate the resulting proteolysis. The proteolytic activity of the most active strain (LH88) was further analyzed (LC-MS/MS and GC-MS) and it was shown that the ß-conglycinin α subunit 1, ß-conglycinin α' subunit, glycinin G1, and 2S albumin were specifically degraded. Peptigram analysis and visualization of the crystal structure showed that the hydrolysis sites of ß-conglycinin α subunit, α' subunit, and the glycinin G1 were located on the surface of the molecule or at the mobile disordered region, hence being highly accessible for the proteinase of LH88. The proteins were partially further degraded to free amino acids, and subsequently catabolized to volatile compounds. However, most of the proteins remained native, even after seven days of fermentation, thus additional modification of protein structure or adjustment of fermentation conditions are required for effective generation of flavor compounds.


Assuntos
Lactobacillus helveticus/metabolismo , Proteínas de Soja/metabolismo , Aminoácidos/análise , Técnicas de Cultura Celular por Lotes , Cromatografia Líquida de Alta Pressão , Cromatografia Gasosa-Espectrometria de Massas , Lactobacillus helveticus/crescimento & desenvolvimento , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/isolamento & purificação , Fragmentos de Peptídeos/metabolismo , Peptídeos/análise , Peptídeos/metabolismo , Proteólise , Proteínas de Armazenamento de Sementes/química , Proteínas de Armazenamento de Sementes/isolamento & purificação , Proteínas de Armazenamento de Sementes/metabolismo , Proteínas de Soja/química , Proteínas de Soja/isolamento & purificação , Espectrometria de Massas em Tandem , Compostos Orgânicos Voláteis/análise
13.
Food Chem ; 340: 127876, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32871354

RESUMO

Jackfruit is a sweet tropical fruit with very pleasant aroma, and the ripe seeds are edible. In this study, jackfruit seed proteins were isolated and subjected to trypsin digestion. The resultant protein hydrolysate was then subjected to antioxidant assay-guided purification, using centrifugal filtration, C18 reverse-phase and strong cation exchange (SCX) fractionations. The purified SCX fraction was further analyzed by de novo peptide sequencing, and two peptide sequences were identified and synthesized. Peptide JFS-2 (VGPWQK) was detected with antioxidant potential, with EC50 value comparable to that of commercial GSH antioxidant peptide. Additionally, the identified peptides were tested with protein protection potential, in an albumin protein denaturation inhibitory assay. Concurrently, we also investigated the pH, temperature, and gastrointestinal-digestion stability profiles for the identified peptide. With further research efforts, the identified peptides could potentially be developed into preservative agent for protein-rich food systems or as health-promoting diet supplements.


Assuntos
Antioxidantes/análise , Artocarpus/química , Peptídeos/análise , Peptídeos/química , Hidrolisados de Proteína/química , Sementes/química , Antioxidantes/química , Cromatografia por Troca Iônica , Digestão , Conservantes de Alimentos , Concentração de Íons de Hidrogênio , Peptídeos/metabolismo , Temperatura , Tripsina
14.
Food Chem ; 334: 127603, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32712492

RESUMO

Present work comprises the use of different solid-state Nuclear Magnetic Resonance strategies for characterizing structural and motional aspects of the peptide matrix that compose a set of four lyophilized Mexican cheese aqueous soluble extracts, each with a controlled ripening. Heteronuclear dipolar coupling modulation schemes allowed to characterize local mobility and structural homogeneity of cheeses' peptide segments in the solid-state as a function of ripening. Results suggest that ripened samples with certain local flexibility but important structural homogeneity present efficient microbial inhibition against tested bacterial strains, whilst high local rigidity of peptides within ripened cheese soluble fractions could partially explain the observed lack of antimicrobial activity. The present study attempts to propose novel observables for lyophilized cheese water soluble extracts that could be partially associated to their ripening-dependent antimicrobial activities, whereas said observables shall contribute to the better targeting, design and optimization of solid-state natural food bio-preservatives.


Assuntos
Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Queijo , Espectroscopia de Ressonância Magnética/métodos , Anti-Infecciosos/análise , Isótopos de Carbono , Queijo/análise , Liofilização , Testes de Sensibilidade Microbiana , Peptídeos/análise , Peptídeos/química , Peptídeos/farmacologia , Solubilidade , Água
15.
Food Chem ; 338: 128029, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-32932089

RESUMO

Peptides derived from whole proteins in beef tenderloin (M. psoas major, PM) and striploin (M. longissimus lumborum, LL) associated with meat quality and muscle fiber composition were identified and quantified during 21 days of aging. Peptide quantification revealed 40-43 proteins to be significantly degraded during all aging time, and these were mostly sarcoplasmic proteins. Cooking loss of both muscles was not changed by aging (P > 0.05), whereas Warner-Bratzler shear force and meat color were affected by aging. Sensory tenderness increased in PM after 14 days of aging (P < 0.05). PM had a higher type I fiber content, whereas LL had a higher type IIX fiber content (P < 0.05), resulting in differences in proteolysis during all aging periods tested. These findings improve our understanding of different biochemical and physicochemical changes in aged meat according to the muscle type.


Assuntos
Armazenamento de Alimentos/métodos , Carne/análise , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Peptídeos/metabolismo , Animais , Bovinos , Cromatografia Líquida de Alta Pressão , Temperatura Baixa , Cor , Qualidade dos Alimentos , Peptídeos/análise , Mapas de Interação de Proteínas , Espectrometria de Massas em Tandem , Fatores de Tempo
16.
Anal Chem ; 93(3): 1393-1400, 2021 01 26.
Artigo em Inglês | MEDLINE | ID: mdl-33373197

RESUMO

In quantitative mass spectrometry imaging (MSI), the gold standard adds a single structural homologue of the target compound at a known concentration to the sample. This internal standard enables to map the detected intensity of the target molecule against an external calibration curve. This approach, however, ignores local noise levels and disproportional ion suppression effects, which might depend on the concentration of the target compound. To overcome these issues, we propose a novel approach that applies several isotopically labeled versions, each at a different concentration, to the sample. This allows creating individual internal calibration curves for every MSI pixel. As proof of principle, we have quantified an endogenous peptide of histone H4 by matrix-assisted laser desorption/ionization-Q-MSI (MALDI-Q-MSI), using a mixture of three isotopically labeled versions. The usage of a fourth label allowed us to compare the gold standard to our multilabel approach. We observed substantial heterogeneity in ion suppression across the tissue, which disclosed itself as varying slopes in the per-pixel regression analyses. These slopes were histology-dependent and differed from each other by up to a factor of 4. The results were validated by liquid chromatography-mass spectrometry (LC-MS), exhibiting a high agreement between LC-MS and MALDI-Q-MSI (Pearson correlation r = 0.87). A comparison between the multilabel and single-label approaches revealed a higher accuracy for the multilabel method when the local target compound concentration differed too much from the concentration of the single label. In conclusion, we show that the multilabel approach provides superior quantitation compared to a single-label approach, in case the target compound is inhomogeneously distributed at a wide concentration range in the tissue.


Assuntos
Histonas/química , Peptídeos/análise , Animais , Colo/química , Colo/metabolismo , Espectrometria de Massas , Suínos
17.
Food Chem ; 345: 128855, 2021 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-33340899

RESUMO

This study attempts to investigate natural angiotensin-I converting enzyme (ACE) inhibitors. Soybean protein isolated (SPI) hydrolysate (SPIH) was prepared by Alcalase from inexpensive SPI, and their ACE inhibitory peptides were obtained via membrane separation, ethanol precipitation and adsorption chromatography enrichment. Activated carbon was more suitable for peptide enrichment than eight macroporous resins. The peptide fraction yielded under optimal conditions (protein-active carbon mass ratio 2:1; adsorption pH 3.0 and time 2 h; desorption time 2 h) exhibited a 10.4 times higher ACE-inhibitory activity than SPIH. Novel peptides IY, YVVF, LVF, WMY, LVLL and FF (hydrophobicity values 10.51-12.87; activity scores 0.2373-0.999) might be the main contributors to SPIH's ACE inhibition. IY had the lowest IC50 (0.53 ± 0.02 µM). YVVF had the greatest affinity (-9.8 kcal/mol) for 2OC2 (ACE's C-domain receptor) via H-bonds. IY and WMY could be potent ACE inhibitors, and their ACE-inhibitory activities unaltered and increased after in vitro gastrointestinal digestion.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/análise , Simulação por Computador , Digestão , Simulação de Acoplamento Molecular , Peptídeos/análise , Peptidil Dipeptidase A/metabolismo , Proteínas de Soja/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Interações Hidrofóbicas e Hidrofílicas , Peptídeos/metabolismo , Peptídeos/farmacologia , Peptidil Dipeptidase A/química , Conformação Proteica
18.
Food Chem ; 345: 128867, 2021 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-33352405

RESUMO

The study explored the use of spent hen, a major egg industry byproduct, as the starting material for preparing antihypertensive peptides. While previous studies were focused mainly on ACE inhibitory (ACEi) peptides, this work also studied peptides with ACE2 upregulating (ACE2u) activity, an emerging target for treating hypertension. Spent hen muscle protein hydrolysate prepared by thermoase (SPH-T) exhibited both ACEi and ACE2u activities. After ultrafiltration and chromatographic fractionation, five potent ACEi peptides, VRP, LKY, VRY, KYKA, and LKYKA, with IC50 values of 0.034-5.77 µg/mL, respectively, and four ACE2u peptides, VKW, VHPKESF, VVHPKESF and VAQWRTKYETDAIQRTEELEEAKKK, which increased ACE2 expression by 0.52-0.84 folds, respectively, were identified; VKW also showed ACEi activity. All peptides, except for VRP, are susceptible to degradation during the simulated gastrointestinal digestion. Our study supports the potential use of spent hens as antihypertensive functional food ingredients and nutraceuticals.


Assuntos
/antagonistas & inibidores , Galinhas , Proteínas Musculares/química , Peptídeos/análise , Peptídeos/isolamento & purificação , Regulação para Cima/efeitos dos fármacos , Sequência de Aminoácidos , Inibidores da Enzima Conversora de Angiotensina/química , Animais , Digestão , Feminino , Hidrólise , Peptídeos/química , Peptídeos/farmacologia
19.
Food Chem ; 339: 128159, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33152898

RESUMO

During production in Chinese baijiu fermentation process, huge amounts of the by-product vinasse are generated and generally utilized as low-value animal feed. We applied alkaline extraction in combination with ultrasonication to recover vinasse proteins, which were then hydrolyzed by complex protease Corolase PP for 8 h to obtain peptide fractions (VPH-1, -2, -3) displaying high DPPH radical scavenging activity. VPH-3 (<3 kDa) separated by ultrafiltration had EC50 values lower than those of VPH-1 and -2 for reactive oxygen species (ROS) and reactive nitrogen species (RNS) radicals, and significantly inhibited production of NO and pro-inflammatory cytokines in LPS-stimulated RAW264.7 macrophage cells. Active peptides and their amino acid sequences were identified by LC-MS/MS analysis, and five synthesized peptides (particularly KLPDHPKLPK and VDVPVKVPYS) displayed strong anti-inflammatory activity at concentration 0.25 mg/mL. These findings will be useful in future commercial development of baijiu vinasse, including application as a new source of bioactive peptides.


Assuntos
Bebidas Alcoólicas , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Peptídeos/farmacologia , Animais , Anti-Inflamatórios não Esteroides/química , Antioxidantes/química , Cromatografia Líquida , Avaliação Pré-Clínica de Medicamentos , Hidrólise , Camundongos , Peptídeos/análise , Peptídeos/química , Proteínas de Plantas/análise , Proteínas de Plantas/farmacologia , Hidrolisados de Proteína/análise , Hidrolisados de Proteína/química , Hidrolisados de Proteína/farmacologia , Células RAW 264.7 , Espécies Reativas de Oxigênio , Espectrometria de Massas em Tandem
20.
Methods Mol Biol ; 2158: 295-305, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32857382

RESUMO

Exosomes are membrane-bound nano-vehicles shed by most eukaryotic cells. Exosomes contain specific proteins and RNAs from parent cells, and they play key signaling roles in cellular development, modulation, and tissue regeneration. Attempts to isolate and modify exosomes to increase their targeting efficiency to specific tissue are still in their infancy. Here, we describe generation of exosomes from biopsy, isolation of exosomes by centrifugal ultrafiltration method, and approaches for manipulation of cardiac homing exosomes by chemical engineering for the treatment of myocardial infarction.


Assuntos
Exossomos , Tecnologia Farmacêutica/métodos , Animais , Plaquetas/química , Técnicas de Cultura de Células/métodos , Fracionamento Celular/métodos , Membrana Celular/química , Membrana Celular/metabolismo , Exossomos/ultraestrutura , Humanos , Camundongos , Terapia de Alvo Molecular , Miocárdio/citologia , Peptídeos/análise , Peptídeos/metabolismo , Ratos , Ultracentrifugação , Ondas Ultrassônicas
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...