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1.
Mar Drugs ; 22(4)2024 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-38667757

RESUMO

Hypertension, a major health concern linked to heart disease and premature mortality, has prompted a search for alternative treatments due to side effects of existing medications. Sustainable harvesting of low-trophic marine organisms not only enhances food security but also provides a variety of bioactive molecules, including peptides. Despite comprising only a fraction of active natural compounds, peptides are ideal for drug development due to their size, stability, and resistance to degradation. Our review evaluates the anti-hypertensive properties of peptides and proteins derived from selected marine invertebrate phyla, examining the various methodologies used and their application in pharmaceuticals, supplements, and functional food. A considerable body of research exists on the anti-hypertensive effects of certain marine invertebrates, yet many species remain unexamined. The array of assessments methods, particularly for ACE inhibition, complicates the comparison of results. The dominance of in vitro and animal in vivo studies indicates a need for more clinical research in order to transition peptides into pharmaceuticals. Our findings lay the groundwork for further exploration of these promising marine invertebrates, emphasizing the need to balance scientific discovery and marine conservation for sustainable resource use.


Assuntos
Anti-Hipertensivos , Organismos Aquáticos , Suplementos Nutricionais , Alimento Funcional , Invertebrados , Peptídeos , Animais , Humanos , Anti-Hipertensivos/farmacologia , Organismos Aquáticos/química , Produtos Biológicos/farmacologia , Hipertensão/tratamento farmacológico , Invertebrados/química , Peptídeos/análise , Peptídeos/farmacologia
2.
Anal Bioanal Chem ; 416(14): 3349-3360, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38607384

RESUMO

The analysis of almost holistic food profiles has developed considerably over the last years. This has also led to larger amounts of data and the ability to obtain more information about health-beneficial and adverse constituents in food than ever before. Especially in the field of proteomics, software is used for evaluation, and these do not provide specific approaches for unique monitoring questions. An additional and more comprehensive way of evaluation can be done with the programming language Python. It offers broad possibilities by a large ecosystem for mass spectrometric data analysis, but needs to be tailored for specific sets of features, the research questions behind. It also offers the applicability of various machine-learning approaches. The aim of the present study was to develop an algorithm for selecting and identifying potential marker peptides from mass spectrometric data. The workflow is divided into three steps: (I) feature engineering, (II) chemometric data analysis, and (III) feature identification. The first step is the transformation of the mass spectrometric data into a structure, which enables the application of existing data analysis packages in Python. The second step is the data analysis for selecting single features. These features are further processed in the third step, which is the feature identification. The data used exemplarily in this proof-of-principle approach was from a study on the influence of a heat treatment on the milk proteome/peptidome.


Assuntos
Temperatura Alta , Leite , Peptídeos , Fluxo de Trabalho , Leite/química , Animais , Peptídeos/análise , Peptídeos/química , Biomarcadores/análise , Software , Proteômica/métodos , Espectrometria de Massas/métodos , Linguagens de Programação , Algoritmos
3.
Electrophoresis ; 45(1-2): 120-164, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37705480

RESUMO

Therapeutic peptides are a growing class of innovative drugs with high efficiency and a low risk of adverse effects. These biomolecules fall within the molecular mass range between that of small molecules and proteins. However, their inherent instability and potential for degradation underscore the importance of reliable and effective analytical methods for pharmaceutical quality control, therapeutic drug monitoring, and compliance testing. Liquid chromatography-mass spectrometry (LC-MS) has long time been the "gold standard" conventional method for peptide analysis, but capillary electrophoresis (CE) is increasingly being recognized as a complementary and, in some cases, superior, highly efficient, green, and cost-effective alternative technique. CE can separate peptides composed of different amino acids owing to differences in their net charge and size, determining their migration behavior in an electric field. This review provides a comprehensive overview of therapeutic peptides that have been used in the clinical environment for the last 25 years. It describes the properties, classification, current trends in development, and clinical use of therapeutic peptides. From the analytical point of view, it discusses the challenges associated with the analysis of therapeutic peptides in pharmaceutical and biological matrices, as well as the evaluation of CE as a whole and the comparison with LC methods. The article also highlights the use of microchip electrophoresis, nonaqueous CE, and nonconventional hydrodynamically closed CE systems and their applications. Overall, the article emphasizes the importance of developing new CE-based analytical methods to ensure the high quality, safety, and efficacy of therapeutic peptides in clinical practice.


Assuntos
Peptídeos , Proteínas , Peptídeos/análise , Proteínas/análise , Eletroforese Capilar/métodos , Aminoácidos , Preparações Farmacêuticas
4.
Front Immunol ; 14: 1291666, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38077382

RESUMO

Background: Allergens from Fagales trees frequently cause spring allergy in Europe, North America, and some parts of Asia. The definition of the birch homologous group, which includes birch (Bet v), oak (Que a), alder (Aln g), hazel (Cor a), hornbeam (Car b), beech (Fag s), and chestnut (Cas s), is based on high allergen sequence identity and extensive IgE cross-reactivity. Clinical effect was seen during the alder/hazel, birch, and oak pollen seasons after treatment with tree SLIT-tablets containing only birch allergen extract. Here, we characterize T-cell reactivity with respect to epitope specificities and cross-reactivity toward various Bet v 1 family members, (PR-10/group 1 major allergens). This cross-reactivity may be part of the immunological basis of clinical effect or cross-protection when exposed to birch homologous tree species. Method: T-cell lines were generated from 29 birch-allergic individuals through stimulation of peripheral blood mononuclear cells (PBMCs) with birch/Bet v or oak/Que a allergen extracts. T-cell responses to allergen extracts, purified group 1 allergens, and overlapping 20-mer peptides (Bet v 1, Aln g 1, Cor a 1, and Que a 1) were investigated by T-cell proliferation and cytokine production. Cross-reactivity was evaluated based on Pearson's correlations of response strength and further investigated by flow cytometry using tetramer staining for homologous peptide pairs. Results: T-cell reactivity toward extracts and group 1 allergens from across the birch homologous group was observed for birch/Bet v as well as oak/Que a T-cell lines. T-cell lines responded to multiple Bet v 1 homologous peptides from Aln g 1 and Cor a 1 and a subset of Que a 1 peptides. Significant Pearson's correlations between frequently recognized peptides derived from Bet v 1 and the corresponding peptides derived from alder, hazel, and oak strongly supported the T-cell cross-reactivity toward these allergens. Cross-reactivity between birch and birch homologous peptides was confirmed by pMHCII tetramer staining. Conclusion: T cells from birch tree pollen allergic individuals respond to multiple trees within the birch homologous group in accordance with the level of sequence homology between Bet v 1 family members, (PR-10 allergens) from these allergen sources, confirming the basis for clinical cross-protection.


Assuntos
Hipersensibilidade , Árvores , Humanos , Linfócitos T , Leucócitos Mononucleares , Antígenos de Plantas , Pólen , Alérgenos , Peptídeos/análise , Betula
5.
J Agric Food Chem ; 71(11): 4625-4637, 2023 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-36892038

RESUMO

In this study, five novel Se-enriched antioxidant peptides (FLSeML, LSeMAAL, LASeMMVL, SeMLLAA, and LSeMAL) were purified and identified from Se-enriched Moringa oleifera (M. oleifera) seed protein hydrolysate. The five peptides showed excellent cellular antioxidant activity, with respective EC50 values of 0.291, 0.383, 0.662, 0.1, and 0.123 µg/mL. The five peptides (0.025 mg/mL) increased the cell viability from 78.72 to 90.71, 89.16, 93.92, 83.68, and 98.29%, respectively, effectively reducing reactive oxygen species accumulation and significantly increasing superoxide dismutase and catalase activities in damaged cells. Molecular docking results revealed that the five novel Se-enriched peptides interacted with the key amino acid of Keap1, thus directly blocking the interaction of Keap1-Nrf2 and activating the antioxidant stress response to enhance the ability of scavenging free radicals in vitro. In conclusion, Se-enriched M. oleifera seed peptides exhibited significant antioxidant activity and can be expected to find widespread use as a highly active natural functional food additive and ingredient.


Assuntos
Moringa oleifera , Selênio , Antioxidantes/química , Proteína 1 Associada a ECH Semelhante a Kelch , Moringa oleifera/química , Selênio/análise , Simulação de Acoplamento Molecular , Extratos Vegetais/química , Fator 2 Relacionado a NF-E2/análise , Peptídeos/farmacologia , Peptídeos/análise , Sementes/química
6.
J Sep Sci ; 46(9): e2200949, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36821105

RESUMO

Saiga antelope horn and Rhinoceros horn have been used in traditional Chinese medicine for thousands of years. However, due to the protection of wildlife, the application of these rare animal horns has been restricted or prohibited. Therefore, water buffalo horn, goat horn, and yak horn have been applied as alternatives to Rhinoceros horn or Saiga antelope horn in a clinic. It is extremely difficult to distinguish normal animal horns in powdered or decocted form, especially identifying related species such as water buffalo horn, yak horn, and cattle horn. In this work, mathematics set and label-free proteomics analysis were combined for discovering keratin-derived specific peptide biomarkers. By using mathematics set analysis after nano liquid chromatography-tandem mass spectrometry-based proteomics, the selected species-specific peptides could be used to identify the authenticity of the Saiga antelope horn and goat horn. Furthermore, peptide biomarkers were selected to distinguish related species-derived horns, water buffalo horn, yak horn, and cattle horn. In total, eight peptide biomarkers were selected and applied for simultaneously distinguishing different horn samples. The present strategy provides a method for peptide biomarkers discovery and also has positive significance for ensuring the quality and efficacy of animal horn-derived traditional Chinese medicines and their products.


Assuntos
Antílopes , Cornos , Animais , Bovinos , Medicina Tradicional Chinesa , Queratinas , Búfalos , Proteômica , Cornos/química , Peptídeos/análise , Perissodáctilos , Cabras , Biomarcadores/análise , Matemática
7.
J Chromatogr A ; 1688: 463692, 2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36549145

RESUMO

Panax ginseng is a precious and ancient medicinal plant. The completion of its genome sequencing has laid the foundation for the study of proteome and peptidome. However, the high abundance of secondary metabolites in ginseng reduces the identification efficiency of proteins and peptides in mass spectrometry. In this report, strong cation exchange pretreatment was carried out to eliminate the interference of impurities. Based on the charge separation of proteolytic peptides and metabolites, the sensitivity of mass spectrometry detection was greatly improved. After pretreatment, 2322 and 2685 proteins were identified from the root and stem leaf extract. Further, the ginseng peptidome was analyzed based on this optimized strategy, where 970 and 653 endogenous peptides were identified from root and stem leaf extract, respectively. Functional analysis of proteins and endogenous peptides provided valuable information on the biological activities, metabolic processes, and ginsenoside biosynthesis pathways of ginseng.


Assuntos
Ginsenosídeos , Panax , Panax/química , Proteômica , Espectrometria de Massas , Cromatografia Líquida , Ginsenosídeos/análise , Extratos Vegetais/química , Peptídeos/análise , Raízes de Plantas/química , Cromatografia Líquida de Alta Pressão
8.
Molecules ; 27(17)2022 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-36080442

RESUMO

The chemical substances responsible for the kokumi taste of green tea infusion are still unclear. Here, we isolated the kokumi compound-containing fractions from green tea infusion through ultrafiltration, and the major kokumi compounds were characterized as γ-Glu-Gln and γ-Glu-Cys-Gly (GSH) through ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS). The results indicated that peptides and amino acids were essential compounds in the kokumi-enriched fractions for conducting the sense of kokumi. L-theanine had an enhancing effect on the kokumi taste of green tea infusion, which was confirmed in the sensory reconstitution study. Thus, peptides, especially γ-Glu-Gln and GSH, are the major kokumi compounds in green tea infusion, which has the potential of improving the flavor of tea beverages.


Assuntos
Camellia sinensis , Camellia sinensis/química , Cromatografia Líquida de Alta Pressão , Peptídeos/análise , Paladar , Chá/química
9.
Nutrients ; 14(18)2022 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-36145113

RESUMO

Antler bone calcium (AB-Ca) and bioactive peptides (ABPs) were extracted from antler bones (Cervus elaphus) to maximize their value. In this study, 0.14 g calcium was obtained from 1 g antler bone. The peptide-calcium chelate rate was 53.68 ± 1.80%, and the Gly, Pro, and Glu in ABPs were identified to donate most to the increased calcium affinity through the mass spectrometry. Fourier transform infrared spectroscopy showed that calcium predominantly interacted with amino nitrogen atoms and carboxyl oxygen atoms, thereby generating a peptide-calcium chelate. The peptide-calcium chelates were characterized using scanning electron microscopy. A Caco-2 cell monolayer model showed that ABPs significantly increased calcium transport. Furthermore, the D-gal-induced aging mouse model indicated that the ABPs + AB-Ca group showed higher Ca and PINP levels, lower P, ALP, and CTX-1content in serum, and considerably higher tibia index and tibia calcium content. Results showed that ABPs + AB-Ca increased bone formation and inhibited bone resorption, thereby providing calcium supplements for ameliorating senile osteoporosis (SOP).


Assuntos
Chifres de Veado , Cervos , Envelhecimento , Animais , Chifres de Veado/química , Células CACO-2 , Cálcio/análise , Cálcio da Dieta/análise , Modelos Animais de Doenças , Humanos , Camundongos , Oxigênio/análise , Peptídeos/análise , Peptídeos/farmacologia
10.
Food Funct ; 13(6): 3481-3494, 2022 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-35246675

RESUMO

Moringa oleifera seed protein hydrolysates exhibit good hypoglycemic activity, but their specific peptide components have not yet been characterized. Here, we identified the ultrafiltration peptide components (<3 kDa) of M. oleifera seed protein hydrolysates. A highly active α-glucosidase inhibitory peptide with an IC50 value of 109.65 µM (MoHpP-2) with the amino acid sequence KETTTIVR was identified. We characterized its structural properties, stability, and hypoglycemic activity. MoHpP-2 was found to be an amphipathic peptide with a ß-turn structure, and the hemolysis of red blood cells was not observed when its concentration was lower than 2 mg mL-1. MoHpP-2 was stable under weakly acidic conditions, at temperatures lower than 60 °C, and at high ion concentrations. Western blotting revealed that MoHpP-2 affected the PI3K and AMPK pathways of HepG2 cells. Molecular docking revealed that MoHpP-2 interacted with α-glucosidase through hydrogen bonding and hydrophobic forces. Thus, MoHpP-2 from M. oleifera seeds could be used to make hypoglycemic functional foods.


Assuntos
Moringa oleifera , Hipoglicemiantes/análise , Hipoglicemiantes/farmacologia , Simulação de Acoplamento Molecular , Moringa oleifera/química , Peptídeos/análise , Peptídeos/farmacologia , Extratos Vegetais/análise , Extratos Vegetais/farmacologia , Hidrolisados de Proteína/farmacologia , Sementes/química
11.
Sci Rep ; 12(1): 1175, 2022 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-35064192

RESUMO

Intact (whole) cell matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) is an established method for biotyping in clinical microbiology as well as for revealing phenotypic shifts in cultured eukaryotic cells. Intact cell MALDI-TOF MS has recently been introduced as a quality control tool for long-term cultures of pluripotent stem cells. Despite the potential this method holds for revealing minute changes in cells, there is still a need for improving the ionization efficiency or peak reproducibility. Here we report for the first time that supplementation by fine particles of black phosphorus to the standard MALDI matrices, such as sinapinic and α-cyano-4-hydroxycinnamic acids enhance intensities of mass spectra of particular amino acids and peptides, presumably by interactions with aromatic groups within the molecules. In addition, the particles of black phosphorus induce the formation of small and regularly dispersed crystals of sinapinic acid and α-cyano-4-hydroxycinnamic acid with the analyte on a steel MALDI target plate. Patterns of mass spectra recorded from intact cells using black phosphorus-enriched matrix were more reproducible and contained peaks of higher intensities when compared to matrix without black phosphorus supplementation. In summary, enrichment of common organic matrices by black phosphorus can improve discrimination data analysis by enhancing peak intensity and reproducibility of mass spectra acquired from intact cells.


Assuntos
Fósforo/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Aminoácidos/análise , Aminoácidos/química , Técnicas de Cultura de Células/métodos , Linhagem Celular , Células-Tronco Embrionárias Humanas , Humanos , Peptídeos/análise , Peptídeos/química , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/normas
12.
Bioorg Chem ; 116: 105362, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34598089

RESUMO

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC50 (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z' factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.


Assuntos
Alcenos/análise , Antivirais/análise , Polarização de Fluorescência , Ensaios de Triagem em Larga Escala , Peptídeos/análise , Polifenóis/análise , Alcenos/farmacologia , Antivirais/farmacologia , Avaliação Pré-Clínica de Medicamentos , Humanos , Estrutura Molecular , Peptídeos/farmacologia , Polifenóis/farmacologia , SARS-CoV-2/efeitos dos fármacos
13.
Anal Bioanal Chem ; 413(30): 7609-7623, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34668043

RESUMO

Therapeutic peptides have an important effect on physiological function and human health, so it is momentous to quantify and detect low levels of these biomolecules in biological samples for treatment and diagnostic purposes. In the present study, an efficient magnetic solid-phase extraction (MSPE) method was developed based on stearic acid-functionalized magnetic hydroxyapatite nanocomposite (MHAP/SA) as a novel and cost-effective adsorbent for extraction of five hypothalamic-related peptides (goserelin, octreotide, triptorelin, somatostatin, and cetrorelix) from biological samples. To characterize the morphology and physicochemical properties of MHAP/SA, Fourier transform infrared spectroscopy (FT-IR), energy-dispersive X-ray spectroscopy (EDS), field emission scanning microscopy (FE-SEM), CHNS elemental analysis, Brunauer-Emmett-Teller (BET), and vibrating sample magnetometry (VSM) were applied. Under optimum conditions, the proposed method (MSPE-HPLC-UV) represented favorable linearity with R2 ≥ 0.9987, suitable intra- and inter-day precisions (RSD ≤ 6.9% and RSD ≤ 8.1%, respectively, n = 3), and limits of detection and quantification in the range of 0.75-1.12 ng mL-1 and 2.50-3.75 ng mL-1, respectively. Eventually, the proposed method was used for the extraction and quantification of target therapeutic peptides in plasma and urine samples, and satisfactory relative recoveries were achieved in the range of 90.6-110.3%.


Assuntos
Durapatita/química , Hipotálamo/química , Nanocompostos/química , Peptídeos/análise , Extração em Fase Sólida/métodos , Ácidos Esteáricos/química , Cromatografia Líquida de Alta Pressão , Humanos , Limite de Detecção , Microscopia Eletrônica de Varredura , Peso Molecular , Peptídeos/sangue , Peptídeos/urina , Reprodutibilidade dos Testes , Análise Espectral/métodos
14.
Nutrients ; 13(8)2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34444931

RESUMO

Over the centuries, humans have traditionally used garlic (Allium sativum L.) as a food ingredient (spice) and remedy for many diseases. To confirm this, many extensive studies recognized the therapeutic effects of garlic bulbs. More recently, black garlic (BG), made by heat-ageing white garlic bulbs, has increased its popularity in cuisine and traditional medicine around the world, but there is still limited information on its composition and potential beneficial effects. In this study, the metabolite profile of methanol extract of BG (BGE) was determined by high-performance liquid chromatography coupled to tandem mass spectrometry in high-resolution mode. Results allowed to establish that BGE major components were sulfur derivatives, saccharides, peptides, organic acids, a phenylpropanoid derivative, saponins, and compounds typical of glycerophospholipid metabolism. Characterization of the BGE action in cancer cells revealed that antioxidant, metabolic, and hepatoprotective effects occur upon treatment as well as induction of maturation of acute myeloid leukemia cells. These results are interesting from the impact point of view of BG consumption as a functional food for potential prevention of metabolic and tumor diseases.


Assuntos
Alho/química , Leucemia Mieloide Aguda/tratamento farmacológico , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Antioxidantes/farmacologia , Cromatografia Líquida de Alta Pressão/métodos , Humanos , Leucemia Mieloide Aguda/patologia , Peptídeos/análise , Raízes de Plantas/química , Polissacarídeos/análise , Saponinas/análise , Especiarias/análise , Enxofre/análise , Espectrometria de Massas em Tandem/métodos , Células U937
15.
Molecules ; 26(6)2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33809348

RESUMO

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.


Assuntos
Análise de Alimentos/métodos , Proteínas de Carne/análise , Proteínas de Plantas/análise , Sequência de Aminoácidos , Animais , Biomarcadores/análise , Cannabis/química , Cromatografia Líquida , Manipulação de Alimentos , Fraude , Temperatura Alta , Peptídeos/análise , Óleos de Plantas/análise , Estabilidade Proteica , Proteômica/métodos , Sementes/química , Especificidade da Espécie , Espectrometria de Massas em Tandem
16.
J Ethnopharmacol ; 270: 113808, 2021 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-33450289

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Moringa oleifera Lam. leaf (MOL), a rich source of protein and phenolics, was traditionally used to treat various diseases including headaches, fevers, sore throat and dyslipidemia. Recently, MOL was reported to possess antioxidant, anti-dyslipidemia and hepato-renal protective activities, indicating that MOL could become a potential agent to improve metabolic disorders associated with hyperuricemia. The antihyperuricemic effect of MOL hydrolysate (MOLH) with high contents of phenolics and peptides remains unknown. AIM OF THE STUDY: The aim of this study is to investigate xanthine oxidase (XO) inhibitory activity of MOLH, to clarify phenolic and peptide profiles of MOLH, and to evaluate possible mechanism underlying the antihyperuricemic effect of MOLH. MATERIALS AND METHODS: MOLH was prepared by enzymatic hydrolysis using commercial trypsin. XO inhibitory activity was determined by XO reaction-UPLC-MS coupling method. The chemical profiles of the phenolic and peptide fractions of MOLH were determined by UPLC-QTOF-MS/MS. The antihyperuricemic effect of MOLH was evaluated in a potassium oxonate-induced hyperuricemic rat model at doses of 200 and 500 mg/kg. Serum uric acid (UA), urea nitrogen, creatinine (CRE), triglyceride (TG), total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol levels, serum XO activity, liver malondialdehyde (MDA) equivalent level, renal tumor necrosis factor-α and interleukin-1ß levels, and protein expression of renal urate-anion transporter 1, glucose transporter 9 and ATP-binding cassette transporter G2 were determined. RESULTS: The phenolic and peptide fractions played key roles in inhibiting XO activity and blocking uric acid production. Five flavonoids and sixteen polypeptides were identified in the phenolic and peptide fractions of MOLH, respectively. MOLH (200 and 500 mg/kg) could effectively reduce the serum UA level of hyperuricemic rats (p < 0.001) by regulation of serum XO activity (p < 0.05 at 200 mg/kg, p < 0.01 at 500 mg/kg) and renal urate transporters. Besides, MOLH could improve metabolic disorders associated with hyperuricemia by its multiple actions on liver MDA (p < 0.001), serum CRE (p < 0.05 at 500 mg/kg) and serum TG (p < 0.001). CONCLUSION: The results provided scientific evidence that MOLH rich in phenolics and peptides ameliorated hyperuricemia and metabolic disorders. This study validated the potential use of MOLH for regulation of hyperuricemia.


Assuntos
Supressores da Gota/farmacologia , Hiperuricemia/tratamento farmacológico , Moringa oleifera/química , Extratos Vegetais/farmacologia , Xantina Oxidase/antagonistas & inibidores , Animais , Creatinina/sangue , Modelos Animais de Doenças , Flavonoides/farmacologia , Supressores da Gota/química , Supressores da Gota/uso terapêutico , Hiperuricemia/induzido quimicamente , Malondialdeído/metabolismo , Transportadores de Ânions Orgânicos/efeitos dos fármacos , Ácido Oxônico/toxicidade , Peptídeos/análise , Peptídeos/química , Fenóis/análise , Fenóis/química , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Folhas de Planta/química , Ratos , Triglicerídeos/sangue , Ácido Úrico/antagonistas & inibidores , Ácido Úrico/sangue
17.
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
18.
Molecules ; 25(24)2020 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-33302391

RESUMO

Solid-state fermentation with food-grade fungal strains can be applied to enhance the bioactive parameters of agro-industrial by-products. Tempe-type fermentation can be adapted to various substrates, but the key factor is the appropriate strain selection. The aim of this study was to compare the potential of Rhizopus strains for obtaining products of improved antioxidant activity from pumpkin oil cake. For this purpose, substances reacting with the Folin-Ciocalteu reagent, with free radical scavenging potential, as well as reducing power were assessed. The effect of the fermentation on the phytate level and inositol phosphate profile in the material was also monitored. The fermentation resulted in the significant enhancement of the antioxidant potential of pumpkin oil cake in the case of all the strains tested, but the most efficient one was R. oligosporus ATCC 64063. During the course of fermentation, the level of phytate in the material decreased (the highest reduction rate was observed in the oil cake fermented with R. oryzae CBS 372.63), while peptides and fungal glucosamine were accumulated. Tempe-type fermentation can be considered as an alternative way of improving the bioactive parameters of pumpkin oil cake and, thanks to the various activities of different Rhizopus strains, it is possible to obtain products of desired parameters.


Assuntos
Cucurbita/química , Fermentação , Manipulação de Alimentos , Microbiologia de Alimentos , Óleos de Plantas/metabolismo , Rhizopus/metabolismo , Antioxidantes/química , Antioxidantes/farmacologia , Análise de Alimentos , Glucosamina/análise , Fosfatos de Inositol/metabolismo , Peptídeos/análise , Proteínas/análise , Especificidade da Espécie
19.
Food Microbiol ; 91: 103492, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32539979

RESUMO

Herein, the effects of grape seed extract (GSE) on the microflora and biochemical changes of container cultured snakehead (Channa argus) fillets during 11 days of chilled storage were investigated. The sensory analysis, the total number of viable colonies, the total amount of volatile basic nitrogen, and k-value analysis revealed that GSE retarded the deterioration of snakehead fillets. The degradation of inosine 5'-monophosphate and the accumulation of inosine and hypoxanthine in the GSE group were slower than these in the control group. Moreover, GSE treatment effectively decreased the accumulation of putrescine, cadaverine, and histamine. Illumina-MiSeq high throughput sequencing results showed that GSE inhibited the growth of Aeromonas on snakehead fillets. Based on the microbial enumeration, sensory analysis, and k-value, GSE prolonged the shelf life of fillets for 3 days, suggesting its potential for snakehead fillets preservation.


Assuntos
Peixes/microbiologia , Conservantes de Alimentos/farmacologia , Extrato de Sementes de Uva/farmacologia , Microbiota/efeitos dos fármacos , Alimentos Marinhos/microbiologia , Aeromonas/efeitos dos fármacos , Aeromonas/crescimento & desenvolvimento , Aminas/análise , Animais , Aquicultura , Microbiologia de Alimentos , Armazenamento de Alimentos , Microbiota/genética , Peptídeos/análise , Alimentos Marinhos/análise
20.
Molecules ; 25(9)2020 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-32349230

RESUMO

This work aimed to study the anti-bacterial, anti-biofilm and anti-oxidant potential effects of low molecular weight (LMW) peptides (Br-p) isolated from burdock (Arctium lappa L.) roots. We conducted a preliminary study to exclude or confirm the antibiotic activity of the LMW peptides fraction of this plant. Br-p were isolated using gel filtration and a 10 kDa cut-off membrane. The obtained peptides were identified by MALDI TOF/TOF. Antibacterial activity was tested against acne strains using diffusion tests, MIC and MBC. The fibroblast cytotoxicity of Br-p was tested, and the selectivity index (SI) value was determined. The fraction of 46 Br-p peptides isolated from burdock root with a molecular weight below 5000 Da and theoretic pI (isoelectric point) of 3.67-11.83 showed a narrow spectrum of activity against Gram-positive acne bacterial strains. One of the Br-p peptides assessed on MALDI RapidDeNovo was LRCDYGRFFASKSLYDPLKKRR cationic peptide. It was analogous to that contained in A. lappa protein, and theoretically it was matched as a peptide with antibiotic nature. Br-p did not show toxicity to fibroblasts in the tested concentration up to 10 mg/mL, obtaining CC50 10 mg/mL. The SI value for the tested Propionibacterium strains ranged from 160 to 320. Finally, an active dressing based on chitosan/alginate/genipin was prepared using freeze-drying. The formed dressing was evaluated for its anti-acne activity. To sum up: preliminary biological studies confirmed the anti-acne properties of the isolated peptide fraction from burdock root and pointed to the possibility of using it to create an active dressing on the skin.


Assuntos
Acne Vulgar/tratamento farmacológico , Antioxidantes/farmacologia , Arctium/química , Peptídeos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Antibacterianos/análise , Antibacterianos/química , Antibacterianos/farmacologia , Antioxidantes/química , Biofilmes/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Radicais Livres , Concentração Inibidora 50 , Microscopia Confocal , Peptídeos/análise , Peptídeos/isolamento & purificação , Peptídeos/toxicidade , Propionibacterium acnes/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Espectrometria de Massas em Tandem
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