RESUMO
Food proteins and peptides are able to exert a variety of well-known bioactivities, some of which are related to well-being and disease prevention in humans and animals. Currently, an active trend in research focuses on chronic inflammation and oxidative stress, delineating their major pathogenetic role in age-related diseases and in some forms of cancer. The present study aims to investigate the potential effects of pseudocereal proteins and their derived peptides on chronic inflammation and oxidative stress. After purification and attribution to protein classes according to classic Osborne's classification, the immune-modulating, antioxidant, and trypsin inhibitor activities of proteins from quinoa (Chenopodium quinoa Willd.), amaranth (Amaranthus retroflexus L.), and buckwheat (Fagopyrum esculentum Moench) seeds have been assessed in vitro. The peptides generated by simulated gastro-intestinal digestion of each fraction have been also investigated for the selected bioactivities. None of the proteins or peptides elicited inflammation in Caco-2 cells; furthermore, all protein fractions showed different degrees of protection of cells from IL-1ß-induced inflammation. Immune-modulating and antioxidant activities were, in general, higher for the albumin fraction. Overall, seed proteins can express these bioactivities mainly after hydrolysis. On the contrary, higher trypsin inhibitor activity was expressed by globulins in their intact form. These findings lay the foundations for the exploitation of these pseudocereal seeds as source of anti-inflammatory molecules.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Fatores Imunológicos/farmacologia , Proteínas de Vegetais Comestíveis/isolamento & purificação , Proteínas de Vegetais Comestíveis/farmacologia , Sementes/química , Amaranthus/química , Anti-Inflamatórios não Esteroides/química , Antioxidantes/farmacologia , Células CACO-2 , Fracionamento Químico , Chenopodium quinoa/química , Fagopyrum/química , Humanos , Fatores Imunológicos/química , Peptídeos/isolamento & purificação , Peptídeos/farmacologia , Proteínas de Vegetais Comestíveis/farmacocinética , Inibidores da Tripsina/química , Inibidores da Tripsina/farmacologiaRESUMO
Adzuki seed ß-vignin, a vicilin-like globulin, has proven to exert various health-promoting biological activities, notably in cardiovascular health. A simple scalable enrichment procedure of this protein for further nutritional and functional studies is crucial. In this study, a simplified chromatography-independent protein fractionation procedure has been optimized and described. The electrophoretic analysis showed a high degree of homogeneity of ß-vignin isolate. Furthermore, the molecular features of the purified protein were investigated. The adzuki bean ß-vignin was found to have a native size of 146 kDa, and the molecular weight determined was consistent with a trimeric structure. These were identified in two main polypeptide chains (masses of 56-54 kDa) that are glycosylated polypeptides with metal binding capacity, and one minor polypeptide chain with a mass 37 kDa, wherein these features are absent. The in vitro analysis showed a high degree of digestibility of the protein (92%) and potential anti-inflammatory capacity. The results lay the basis not only for further investigation of the health-promoting properties of the adzuki bean ß-vignin protein, but also for a possible application as nutraceutical molecule.
Assuntos
Cromatografia/métodos , Proteínas de Plantas/genética , Vigna/química , Sequência de Aminoácidos , Células CACO-2 , Fracionamento Químico , Farinha , Globulinas/química , Humanos , Concentração de Íons de Hidrogênio , Inflamação/patologia , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Sementes/química , SolubilidadeRESUMO
γ-conglutin (γC) is a major protein of Lupinus albus seeds, but its function is still unknown. It shares high structural similarity with xyloglucan-specific endo-glucanase inhibitor proteins (XEGIPs) and, to a lesser extent, with Triticum aestivum endoxylanase inhibitors (TAXI-I), active against fungal glycoside hydrolases GH12 and GH11, respectively. However, γC lacks both these inhibitory activities. Since ß-galactomannans are major components of the cell walls of endosperm in several legume plants, we tested the inhibitory activity of γC against a GH2 ß-mannosidase (EC 3.2.1.25). γC was actually able to inhibit the enzyme, and this effect was enhanced by the presence of zinc ions. The stoichiometry of the γC/enzyme interaction was 1:1, and the calculated Ki was 1.55 µM. To obtain further insights into the interaction between γC and ß-mannosidase, an in silico structural bioinformatic approach was followed, including some docking analyses. By and large, this work describes experimental findings that highlight new scenarios for understanding the natural role of γC. Although structural predictions can leave space for speculative interpretations, the full complexity of the data reported in this work allows one to hypothesize mechanisms of action for the basis of inhibition. At least two mechanisms seem plausible, both involving lupin-γC-peculiar structures.
Assuntos
Glucanos/química , Glicosídeo Hidrolases/genética , Lupinus/química , Proteínas de Plantas/genética , Xilanos/química , Sequência de Aminoácidos/genética , Glucanos/genética , Glicosídeo Hidrolases/antagonistas & inibidores , Proteínas de Plantas/ultraestrutura , Proteínas de Armazenamento de Sementes/genética , Proteínas de Armazenamento de Sementes/ultraestrutura , Sementes/química , Sementes/crescimento & desenvolvimento , Triticum/química , Xilanos/genéticaRESUMO
This study was aimed at characterizing the anthocyanins and phenolics profile in different varieties of pigmented corn and wheat and in some of their milling fractions. Acid/ethanol extracts were used to assess total anthocyanins, overall antioxidant activity, the overall polyphenol profile, and for evaluating the inhibition of pancreatic α-amylase and of intestinal α-glucosidase. Both enzymes were inhibited in a dose-dependent manner by all extracts, but individual extracts had specific effects on each enzyme. Anti-inflammatory response was evaluated by using acid-free extracts and Caco-2 cells transiently transfected with a luciferase reporter gene responding to cytokine stimulation. The immune response of interleukin-stimulated cells decreased significantly in a dose-dependent manner in the presence of 20-50 µM/l anthocyanins from all grains extracts, again with a different efficiency. The inhibitory ability and the anti-inflammatory capability of these extracts are in most cases higher than in similar extracts from other sources, suggesting that activities in each extract may imply specific synergies between anthocyanins and other phenolics.
Assuntos
Antocianinas/farmacologia , Grão Comestível/química , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Triticum/química , Zea mays/química , Antocianinas/análise , Antioxidantes/metabolismo , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Alimento Funcional , Inibidores de Glicosídeo Hidrolases/análise , Inibidores de Glicosídeo Hidrolases/farmacologia , Humanos , Intestinos/enzimologia , Pâncreas/enzimologia , Fenóis/análise , Pigmentos Biológicos/análise , Pigmentos Biológicos/farmacologia , Extratos Vegetais/química , Polifenóis/análise , Polifenóis/farmacologia , alfa-Amilases/antagonistas & inibidores , alfa-Amilases/metabolismo , alfa-Glucosidases/metabolismoRESUMO
The basic 7S globulin (Bg7S) is one of the major globulins of soybean seeds. Despite its dual subunit composition and oligomeric assembly, Bg7S has a compact 3D structure (PDB: 3AUP) which is stabilized by a network of inter- and intra-chain disulphide bridges. Bg7S shares several structural elements with a number of homologous proteins from other seeds, whose function is still uncertain. In this work, Bg7S native conformation was probed by using the proteolytic enzyme trypsin. In spite of the presence of many arginine and lysine residues, the protein resulted extremely recalcitrant to in vitro enzymatic cleavage. Indeed, only two scissile bonds located near the C- and N-termini of the large and small subunits, respectively, were cleaved. The partially cleaved products were stable even at prolonged incubation times. Although the generated small peptide fragments were not covalently bound to the remnant of the main chains, they were held in place, as assessed by denaturing and non-denaturing chromatographic approaches. Moreover, both the already observed pH-dependent association/dissociation behaviour of the protein and its insulin binding capacity were preserved both at neutral and acidic pH values. These results are in line with the growing view that the degradation of seed proteins, either storage and non-storage, may be a controlled process related to specific functionalities.
Assuntos
Globulinas/química , Glycine max/química , Técnicas de Sonda Molecular , Sementes/química , Proteínas de Soja/química , Tripsina/química , Sítios de Ligação , Modelos Químicos , Modelos Moleculares , Sondas Moleculares/química , Ligação Proteica , Conformação ProteicaRESUMO
Cowpea seed ß-vignin, a vicilin-like globulin, proved to exert various health favourable effects, including blood cholesterol reduction in animal models. The need of a simple scalable enrichment procedure for further studies for tailored applications of this seed protein is crucial. A chromatography-independent fractionation method allowing to obtain a protein preparation with a high degree of homogeneity was used. Further purification was pursued to deep the molecular characterisation of ß-vignin. The results showed: (i) differing glycosylation patterns of the two constituent polypeptides, in agreement with amino acid sequence features; (ii) the seed accumulation of a gene product never identified before; (iii) metal binding capacity of native protein, a property observed only in few other legume seed vicilins.
Assuntos
Globulinas/química , Globulinas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Vigna/química , Glicosilação , Metais/metabolismo , Sementes/químicaRESUMO
The purpose of this study was to observe, in a rat animal model, the short and medium term effects of vegan (VEG) or omnivorous (OMNI) diets with different energy partition between nutrients (zone or classic). Six different diets were administered, for 72 days to 120 growing male Sprague-Dawley rats: (i) VEG zone diet; (ii) VEG classic diet; (iii) OMNI zone diet; (iv) OMNI classic diet; (v) OMNI zone diet with added fibre and (vi) OMNI classic diet with added fibre. Zone diets (high protein and low carbohydrates), resulted in better growth , feed efficiency, lower blood glucose and insulin responses. VEG diets have lowered cholesterol blood level. Histopathological analysis evidenced no damage to liver and kidney tissue by the intake of any of the diet types. Further longer animal and human duration studies should be performed to exclude detrimental effect of higher protein diet.
Assuntos
Ração Animal/análise , Dieta Vegana , Carboidratos da Dieta/administração & dosagem , Proteínas Alimentares/administração & dosagem , Metabolismo Energético/efeitos dos fármacos , Tecido Adiposo , Animais , Fibras na Dieta , Ingestão de Energia , Epididimo , Masculino , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
Fermentation represents a valuable and cost-effective approach for food stabilisation and nutritional improvement. Tempeh is an example of soybean solid-state fermentation. In this work, we investigated the possibility of producing a tempeh analogue containing high amounts of vitamin B12 using seeds of three different species of the legume lupin, namely Lupinus albus, L. angustifolius and L. mutabilis, with Rhizopus oligosporus and Propionibacterium freudenreichii cofermentation. Synergic effects of Rhizopus and Propionibacterium in increasing vitamin B12 up to 1230 ng/g dw was observed. These findings indicate that this cofermentation can improve lupin nutritional quality and safety to provide a tempeh analogue with added value for vegan and vegetarian communities and low-income populations. The level of potentially toxic lupin alkaloids was also monitored during the tempeh preparation.
Assuntos
Lupinus , Propionibacterium/metabolismo , Rhizopus/metabolismo , Alimentos de Soja/análise , Vitamina B 12/química , Fermentação , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Vitamina B 12/metabolismoRESUMO
Lupin seed γ-Conglutin is a protein capable of reducing glycaemia in mammalians and increasing glucose uptake by model cells. This work investigated whether γ-Conglutin is internalised into the target cells and undergoes any covalent change during the process, as a first step to understanding its mechanism of action. To this purpose, γ-Conglutin-treated and untreated HepG2 cells were submitted to confocal and transmission electron microscopy. Immune-revelation of γ-Conglutin at various intervals revealed its accumulation inside the cytosol. In parallel, 2D-electrophoresis of the cell lysates and antibody reaction of the blotted maps showed the presence of the protein intact subunits inside the treated cells, whilest no trace of the protein was found in the control cells. However, γ-Conglutin-related spots with an unexpectedly low pI were also observed in the maps. These spots were excised, trypsin-treated and submitted to MS/MS spectrometric analysis. The presence of phosphorylated amino acids was detected. These findings, by showing that γ-Conglutin is internalised by HepG2 cells in an intact form and is modified by multiple phosphorylation, open the way to the understanding of the lupin γ-Conglutin insulin-mimetic activity.
Assuntos
Hipoglicemiantes/metabolismo , Lupinus/química , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Células Hep G2 , Humanos , Dados de Sequência Molecular , Fosforilação , Sementes/químicaRESUMO
BACKGROUND: Ochratoxin A (OTA) is a mycotoxin present in food that can be found in human blood, due to its long half-life. Plasma OTA detection represents a good parameter for evaluating the exposure at the population level. PURPOSE: The relation between plasma OTA levels, dietary habits, and specific disease risk biomarkers (body mass index (BMI), C-reactive protein (CRP), and cardiovascular risk score) was investigated. METHODS: The study involved 327 subjects (150 men and 177 women) aged between 38 and 48 years. Food consumption was evaluated by means of the EPIC questionnaire; plasma OTA was measured by HPLC; CRP was determined in fresh serum samples by a latex particle-enhanced immunoturbidimetric assay. RESULTS: OTA was detected in 99.1% of plasma samples (LOD 25 ng/L); the mean ± SD value was 0.229 ± 0.238 ng/mL. However, only 5.2% of samples exceeded 500 ng/L, considered the threshold for a possible pathogenic activity. The estimated mean daily dietary intake of OTA resulted 0.452 ± 0.468 ng/kg body weight (bw)/day, markedly lower than the tolerable daily intake set by EFSA (17.1 ng/kg bw/day). Processed and mutton/lamb meat were found to contribute most to plasma OTA variance. Nevertheless, cereals, wine, beer, and jam/honey consumption correlated positively with OTA levels. Plasma OTA showed a significant positive association with CRP and cardiovascular risk score (ß = 0.20 ± 0.08; P = 0.015 and ß = 0.25 ± 0.08; P = 0.001, respectively); however, the association was present in men but not in women. CONCLUSIONS: Even if the hypothesis of a possible hepatic toxicity of OTA in humans is yet to be verified, the positive association between plasma OTA and CRP may indicate a possible role of OTA in inflammation status and consequently in the genesis of cardiovascular diseases and cancer.
Assuntos
Biomarcadores/sangue , Exposição Ambiental/efeitos adversos , Contaminação de Alimentos/análise , Ocratoxinas/sangue , Adulto , Cerveja/análise , Índice de Massa Corporal , Proteína C-Reativa/análise , Doenças Cardiovasculares/epidemiologia , Cromatografia Líquida de Alta Pressão , Grão Comestível/química , Comportamento Alimentar , Feminino , Meia-Vida , Humanos , Itália/epidemiologia , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Fatores de Risco , Inquéritos e Questionários , Vinho/análiseRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson (syn. A. moschata Wulfen) (Asteraceae) is an alpine endemic plant whose aerial parts are harvested by the locals mainly for the digestive properties. Despite its widespread use, few studies have been conducted to date to verify its bioactivity. AIM OF THE STUDY: The purpose of the work was to meet the tradition confirming with experimental data the popular belief that the consumption of this species offers beneficial effects to the gastrointestinal system. MATERIALS AND METHODS: Using Soxhlet apparatus, the dried aerial parts of A. erba-rotta subsp. moschata were successively extracted with petroleum ether (PET), dichloromethane (DCM) and methanol (MeOH). The essential oil (EO) was obtained by hydrodistillation using a Clevenger apparatus while infusion (AE) was prepared following the traditional local recipe. Their chemical characterization was performed by various techniques including SPME-GC/MS, GC/MS and HPLC/MS-MS. An in vitro biological screening was carried out. The influence of AE on lipid digestion was monitored by titration of free fatty acids (FFA) during pancreatic lipase activity with the pH-stat method. For all extracts and EO, the anti-Helicobacter pylori activity was assessed by the broth microdilution method, the influence on cell viability was evaluated against NCI-N87, OE21 and Caco-2 cell lines and a preliminary toxicity evaluation was done using Brine Shrimp lethality (BSL) assay. The anti-inflammatory potential was evidenced by interleukin IL-1- induced IL8 expression on Caco-2 cells. RESULTS: AE increased by 15% the FFA releasing compared to the pancreatic lipase alone. PET, DCM and MeOH extracts as well as AE and EO were considered active against the growth of both antimicrobial susceptible and resistant strains of H. pylori with MIC values starting from 16 µg/mL. PET and DCM (IC50 = 89 µg/mL and 96 µg/mL, respectively, against Caco-2 cell line) extracts showed the high effect on cell viability while the EO reduced in 50% of cell viability at 1.48 µL/mL (NCI-N87 cells), 1.42 µL/mL (OE21 cells), and 3.44 µL/mL (Caco-2 cells) corroborating the BSL results. In different degrees, all extracts and EO inhibited the IL-1ß-stimulated IL-8 production in Caco-2 cells. CONCLUSIONS: The obtained data are encouraging and provide a scientific basis for the traditional use of A. erba-rotta subsp. moschata as a digestive agent although they need to be further corroborated by studies involving the investigation of both the in vivo activities and the role of the compounds detected in the extracts.
Assuntos
Achillea , Asteraceae , Óleos Voláteis , Achillea/química , Antioxidantes/farmacologia , Células CACO-2 , Digestão , Humanos , Lipase , Óleos Voláteis/farmacologia , Componentes Aéreos da Planta/química , Extratos Vegetais/químicaRESUMO
Several food-derived molecules, including proteins and peptides, can show bioactivities toward the promotion of well-being and disease prevention in humans. There is still a lack of information about the potential effects on immune and inflammatory responses in mammalian cells following the ingestion of seed storage proteins. This study, for the first time, describes the potential immunomodulation capacity of chenopodin, the major protein component of quinoa seeds. After characterizing the molecular features of the purified protein, we were able to separate two different forms of chenopodin, indicated as LcC (Low charge Chenopodin, 30% of total chenopodin) and HcC (High charge Chenopodin, 70% of total chenopodin). The biological effects of LcC and HcC were investigated by measuring NF-κB activation and IL-8 expression studies in undifferentiated Caco-2 cells. Inflammation was elicited using IL-1ß. The results indicate that LcC and HcC show potential anti-inflammatory activities in an intestinal cell model, and that the proteins can act differently, depending on their structural features. Furthermore, the molecular mechanisms of action and the structural/functional relationships of the protein at the basis of the observed bioactivity were investigated using in silico analyses and structural predictions.
Assuntos
Anti-Inflamatórios/farmacologia , Interleucina-1beta/metabolismo , Proteínas de Plantas/farmacologia , Anti-Inflamatórios/química , Sítios de Ligação , Células CACO-2 , Humanos , Interleucina-1beta/química , Interleucina-8/metabolismo , NF-kappa B/metabolismo , Proteínas de Plantas/química , Ligação Proteica , Isoformas de Proteínas/química , Isoformas de Proteínas/farmacologia , Transdução de Sinais/efeitos dos fármacosRESUMO
The search for bioactivities influencing the human wellbeing of food proteins and peptides is a topic of broad and current interest. γ-Conglutin (γC) is a lupin seed protein drawing remarkable pharmacological and/or nutraceutical interest, as it is able to reduce hyperglycemia in humans and animal models. The present work deepens our investigations to understand the molecular basis of the in vitro effects of γC by testing the possible metabolic effects on cultivated Caco-2 cells. γC and its derived peptides (obtained via simulated gastrointestinal digestion) did not influence the cell viability at incubation times up to 24 h. The incubation of cells with native or digested γC caused no detectable inflammation processes mediated by Nuclear Factor kappa B (NFκB). We checked if treatment with γC or its derived peptides can elicit the expression of two peptide transporters (Pept-1 and Htp-1) by using an RT-qPCR approach. Native γC caused the halving of Pept-1 expression compared to untreated cells, but this effect disappeared when γC was digested. Either native γC or γC peptides reduced the expression levels of Hpt-1. Finally, this work also sheds light on the possible structural modifications of γC that may occur in the gastrointestinal tract, using an in vitro simulated dispersed system with polystyrene nanoparticles (NPs).
Assuntos
Tecnologia de Alimentos , Hipoglicemiantes/farmacologia , Lupinus/química , Nanopartículas/química , Proteínas de Plantas/metabolismo , Poliestirenos/química , Adsorção , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Humanos , Hipoglicemiantes/metabolismo , Sementes/química , SoluçõesRESUMO
Interaction with model phospholipid membranes of lupin seed γ-conglutin, a glycaemia-lowering protein from Lupinus albus seeds, has been studied by means of Fourier-Transform infrared spectroscopy at p2H 7.0 and at p2H 4.5. The protein maintains the same secondary structure both at p2H 7.0 and at p2H 4.5, but at p2H 7.0 a higher 1H/2H exchange was observed, indicating a greater solvent accessibility. The difference in Tm and TD1/2 of the protein at the abovementioned p2H's has been calculated around 20⯰C. Infrared measurements have been then performed in the presence of DMPG and DOPA at p2H 4.5. DMPG showed a little destabilizing effect while DOPA exerted a great stabilizing effect, increasing the Tm of γ-conglutin at p2H 4.5 of more than 20⯰C. Since γ-conglutin at p2H 4.5 is in the monomeric form, the interaction with DOPA likely promotes the oligomerization even at p2H 4.5. Interaction between DMPG or DOPA and γ-conglutin has been confirmed by turbidity experiments with DMPC:DMPG or DOPC:DOPA SUVs. Turbidity data also showed high-affinity binding of γ-conglutin to anionic SUVs made up with DOPA. The molecular features outlined in this study are relevant to address the applicative exploitation and to delineate a deeper comprehension of the natural functional role of γ-conglutin.
Assuntos
Bicamadas Lipídicas/metabolismo , Lupinus/metabolismo , Proteínas de Plantas/metabolismo , Medição da Troca de Deutério , Di-Hidroxifenilalanina/química , Concentração de Íons de Hidrogênio , Bicamadas Lipídicas/química , Nefelometria e Turbidimetria , Fosfatidilgliceróis/química , Proteínas de Plantas/química , Sementes/metabolismo , Espectrofotometria Infravermelho , Temperatura de TransiçãoRESUMO
Lupin γ-conglutin and soybean BG7S are two legume seed proteins strongly similar to plant endo-ß-glucanases inhibitors acting against fungal GH11 and GH12 glycoside hydrolase. However these proteins lack inhibitory activity. Here we describe the conversion of lupin γ-conglutin to an active inhibitor of endo-ß-glucanases belonging to GH11 family. A set of γ-conglutin mutants was designed and expressed in Pichia pastoris, along with the wild-type protein. Unexpectedly, this latter was able to inhibit a GH11 enzyme, but not GH12, whereas the mutants were able to modulate the inhibition capacity. In lupin, γ-conglutin is naturally cleaved in two subunits, whereas in P. pastoris it is not. The lack of proteolytic cleavage is one of the reasons at the basis of the inhibitory activity of recombinant γ-conglutin. The results provide new insights about structural features at the basis of the lack of inhibitory activity of wild-type γ-conglutin and its legume homologues.
Assuntos
Celulase/metabolismo , Lupinus/enzimologia , Celulase/química , Celulase/genética , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Lupinus/metabolismo , Mutagênese , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/farmacologiaRESUMO
The 11S storage globulin of white lupin seeds binds to a metal affinity chromatography matrix. Two unusual stretches of contiguous histidine residues, reminiscent of the multiple histidines forming metal binding motifs, at the C-terminal end of 11S globulin acidic chains were hypothesized as candidate elements responsible for the binding capacity. To prove this, the protein was incubated with a lupin seed endopeptidase previously shown to cleave at twin arginine motifs, recurrent in the sequence region of interest. Upon incubation with this enzyme, the loss of metal binding capacity paralleled that of the anti-his-tag reactive polypeptides. The recovered small proteolytic fragment was analyzed by mass spectrometry and N-terminal sequencing and found to correspond to the 24-mer region cleaved off at twin arginine residues and containing the natural his-tag-like region. Similarly, when lupin seeds were germinated for a few days, the his-tag containing 11S globulin chain was converted to a form devoid of such region, suggesting that this mechanism is a part of the natural degradatory process of the protein. The hypothesis that the ordered and controlled dismantling of storage proteins may generate peptide fragments with potential functional roles in plant ontogenesis is presented and discussed.
Assuntos
Globulinas/metabolismo , Lupinus/metabolismo , Proteínas de Plantas/metabolismo , Sementes/metabolismo , Sequência de Aminoácidos , Arginina/química , Arginina/metabolismo , Germinação , Globulinas/química , Lupinus/química , Lupinus/crescimento & desenvolvimento , Modelos Moleculares , Dados de Sequência Molecular , Proteínas de Plantas/química , Proteólise , Sementes/química , Sementes/crescimento & desenvolvimentoRESUMO
The role of seed proteins, especially soyabean 7S globulins, in controlling dyslipidaemia is widely acknowledged. Amino acid sequence homology among the proteins of this family could reflect similar biological functions in other species. The aim of the present study was to unveil a hypolipidaemic effect of the 7S globulins from cowpeas (7S-C) and adzuki beans (7S-A), administered orally to rats fed a hypercholesterolaemic (HC; high cholesterol and TAG) diet for 28 d. A total of forty-five rats were divided into five groups (nine rats per group): (1) standard (STD) diet; (2) HC diet; (3) HC diet + 7S-C (300 mg/kg per d); (4) HC diet + 7S-A (300 mg/kg per d); and (5) HC diet + simvastatin (SVT; 50 mg/kg per d), as a control. Significant decreases in food intake and final body weight of rats receiving HC + 7S-C and HC + 7S-A diets compared with groups fed the HC and STD diets were observed. Significant decreases in serum total and non-HDL-cholesterol of 7S-C, 7S-A and SVT groups were also observed. HDL-cholesterol levels increased in the 7S-C, 7S-A and SVT groups, while hepatic cholesterol and TAG concentrations were significantly lower than in the HC diet group for the 7S-C-supplemented group only. Faecal excretions of fat and cholesterol in HC diet groups were considerably higher in animals consuming the 7S globulins. The results show that cowpea and adzuki bean 7S globulins promote cholesterol-decreasing effects in hypercholesterolaemic rats even at low dosages, as already observed for other legume seed storage proteins of this family. This main effect is discussed in relation to the possible mechanisms of action.
RESUMO
The supplementation of foods with biologically active compounds can be a powerful approach for improving diet and well being. In this study we separately included in pasta matrices a concentrate of γ-conglutin, a glucose-lowering protein from Lupinus albus seeds, an isolate of the other main lupin storage proteins and ovalbumin, at a ratio corresponding to 125 mg of pure protein in 100 g of pasta. With these products we fed rats made hyperglycaemic, for 3 weeks. Among the most relevant changes measured in body and blood parameters were: (i) a significant reduction in food intake of rats fed γ-conglutin concentrate supplemented pasta and a significant limitation in the body weight increase in rats fed α, ß and δ-conglutin isolate supplemented pasta, while the food conversion indices were unchanged; (ii) a reduction in glycaemia upon glucose overload trial, especially in the γ-conglutin concentrate supplemented pasta fed animals, at a dose of 45 mg per kg body weight. The correlations among the measured parameters are discussed. Overall, the results evidence the potentiality of supplementing traditional foods with exogenous nutraceutical seed proteins to control body weight gain and glycaemia.
Assuntos
Glicemia/metabolismo , Aditivos Alimentares/metabolismo , Hiperglicemia/dietoterapia , Lupinus/metabolismo , Proteínas de Plantas/metabolismo , Aumento de Peso , Animais , Ingestão de Alimentos , Humanos , Hiperglicemia/metabolismo , Hiperglicemia/fisiopatologia , Lupinus/química , Masculino , Ratos , Ratos Sprague-Dawley , Proteínas de Armazenamento de Sementes/metabolismo , TriticumRESUMO
Mice fed long-term high-fat diets (HFD) are an established model for human metabolic disorders, such as obesity and diabetes. However, also the effects of short-term HFD feeding should be investigated to understand which are the first events that trigger the onset of a pre-disease condition, the so-called metabolic syndrome, that increases the risk of developing clinical diseases. In this study, C57BL/6N mice were fed a control diet (CTR) or a HFD for 1 (T1) or 2 weeks (T2). Metabolic and histological effects were examined. Cecum transcriptomes of HFD and CTR mice were compared at T2 by microarray analysis. Differentially expressed genes were validated by real-time PCR in the cecum and in the liver. After 2 weeks of diet administration, HFD mice showed an altered expression pattern in only seven genes, four of which are involved in the circadian clock regulatory pathway. Real-time PCR confirmed microarray results of the cecum and revealed the same trend of clock gene expression changes in the liver. These findings suggest that clock genes may play an important role in early controlling gut output systems in response to HFD in mice and that their expression change may also represent an early signaling of the development of an intestinal pro-inflammatory status.
RESUMO
γ-Conglutin is a blood glucose-lowering protein purified from lupin (Lupinus albus, L.) seed. Despite various features of this protein have already been studied, no function in the seed nor any mechanism of action as a hypoglycemic nutraceutical compound have been identified so far. The lupin protein was shown to exist both in monomeric and multimeric forms as a function of pH. However, a detailed description of the pH-dependent structural dynamics of this protein, as the basis to investigate the reason/s of its functional behaviour, is not available yet. In this study, multiple and independent spectroscopic approaches, including light scattering associated to size exclusion chromatography of both untreated and covalently cross-linked protein, near and far UV circular dichroism, intrinsic and extrinsic fluorescence measurements, have been used to monitor oligomeric and conformational modifications caused by pH changes. Altogether, the results revealed a tetramer-dimer-monomer transition between neutral to slightly acidic pH and a dramatic and abrupt conformational change below pH 3.5. According to these findings, a model depicting γ-conglutin structural dynamics was drawn. This model highlights the primary role of amino acid side group electrostatic interactions in the oligomer association/dissociation equilibria and in the pH-driven collapse of the native conformation.