RESUMEN

Textured vegetable proteins (TVP) are an alternative to meet the increasing demand for non-animal food. This study aimed to develop a TVP from mixtures with 45 % pea protein isolate (PPI) enriched with amaranth (AF) and oat (OF) flours using high-moisture extrusion technology (HME) varying the moisture (50-70 %) and the temperature in the second heating zone of the extruder (110-140 °C). After extrusion, all samples demonstrated higher values of water absorption capacity (WAC) than non-extruded mixtures. Mixture of AF:OF:PPI (40:15:45 %) extruded at 60 % moisture and 135 °C showed promising functional properties with WAC and WSI values of 3.2 ± 0.2 g H2O/g and 24.89 ± 2.31 %, respectively, and oil absorption capacity (OAC) of 1.3 g oil/g. The extrusion process altered the thermal and structural properties of proteins promoting a desirable fibrous structure. This confirms the feasibility of using HME to develop TVP based on PPI, AF, and OF.

Asunto(s)

Amaranthus , Avena , Harina , Pisum sativum , Agua , Amaranthus/química , Harina/análisis , Avena/química , Pisum sativum/química , Agua/química , Proteínas de Guisantes/química , Manipulación de Alimentos , Proteínas de Plantas/química , Calor

RESUMEN

Anthocyanins are bioactive compounds responsible for various physiological processes in plants and provide characteristic colors to fruits and flowers. Their biosynthetic pathway is well understood; however, the enzymatic degradation mechanism is less explored. Anthocyanase (ß-glucosidase (BGL)), peroxidase (POD), and polyphenol oxidase (PPO) are enzymes involved in degrading anthocyanins in plants such as petunias, eggplants, and Sicilian oranges. The aim of this work was to investigate the physicochemical interactions between these enzymes and the identified anthocyanins (via UPLC-MS/MS) in cranberry (Vaccinium macrocarpon) through molecular docking to identify the residues likely involved in anthocyanin degradation. Three-dimensional models were constructed using the AlphaFold2 server based on consensus sequences specific to each enzyme. The models with the highest confidence scores (pLDDT) were selected, with BGL, POD, and PPO achieving scores of 87.6, 94.8, and 84.1, respectively. These models were then refined using molecular dynamics for 100 ns. Additionally, UPLC-MS/MS analysis identified various flavonoids in cranberries, including cyanidin, delphinidin, procyanidin B2 and B4, petunidin, pelargonidin, peonidin, and malvidin, providing important experimental data to support the study. Molecular docking simulations revealed the most stable interactions between anthocyanase and the anthocyanins cyanidin 3-arabinoside and cyanidin 3-glucoside, with a favorable ΔG of interaction between -9.3 and -9.2 kcal/mol. This study contributes to proposing a degradation mechanism and seeking inhibitors to prevent fruit discoloration.

Asunto(s)

Antocianinas , Catecol Oxidasa , Simulación del Acoplamiento Molecular , Vaccinium macrocarpon , Antocianinas/química , Antocianinas/metabolismo , Catecol Oxidasa/metabolismo , Catecol Oxidasa/química , Vaccinium macrocarpon/química , Peroxidasa/metabolismo , Peroxidasa/química , Espectrometría de Masas en Tándem , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Simulación de Dinámica Molecular , Simulación por Computador , Frutas/química , Frutas/metabolismo , Frutas/enzimología

RESUMEN

Amidst increasing awareness of diet-health relationships, plant-derived bioactive peptides are recognized for their dual nutritional and health benefits. This study investigates bioactive peptides released after Alcalase hydrolysis of protein from chachafruto (Erythrina edulis), a nutrient-rich South American leguminous plant, focusing on their behavior during simulated gastrointestinal digestion. Evaluating their ability to scavenge radicals, mitigate oxidative stress, and influence immune response biomarkers, this study underscores the importance of understanding peptide interactions in digestion. The greatest contribution to the antioxidant activity was exerted by the low molecular weight peptides with ORAC values for the <3 kDa fraction of HES, GD-HES, and GID-HES of 0.74 ± 0.03, 0.72 ± 0.004, and 0.56 ± 0.01 (µmol TE/mg protein, respectively). GD-HES and GID-HES exhibited immunomodulatory effects, promoting the release of NO up to 18.52 and 8.58 µM, respectively. The findings of this study highlighted the potential of chachafruto bioactive peptides in functional foods and nutraceuticals, supporting human health through dietary interventions.

Asunto(s)

Antioxidantes , Digestión , Erythrina , Péptidos , Proteínas de Plantas , Hidrólisis , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Péptidos/química , Péptidos/metabolismo , Erythrina/química , Antioxidantes/farmacología , Antioxidantes/química , Antioxidantes/metabolismo , Humanos , Subtilisinas/metabolismo , Subtilisinas/química , Estrés Oxidativo , Tracto Gastrointestinal/metabolismo

RESUMEN

Hydrogen peroxide (H2O2) transport by aquaporins (AQP) is a critical feature for cellular redox signaling. However, the H2O2 permeation mechanism through these channels remains poorly understood. Through functional assays, two Plasma membrane Intrinsic Protein (PIP) AQP from Medicago truncatula, MtPIP2;2 and MtPIP2;3 have been identified as pH-gated channels capable of facilitating the permeation of both water (H2O) and H2O2. Employing a combination of unbiased and enhanced sampling molecular dynamics simulations, we investigated the key barriers and translocation mechanisms governing H2O2 permeation through these AQP in both open and closed conformational states. Our findings reveal that both H2O and H2O2 encounter their primary permeation barrier within the selectivity filter (SF) region of MtPIP2;3. In addition to the SF barrier, a second energetic barrier at the NPA (asparagine-proline-alanine) region that is more restrictive for the passage of H2O2 than for H2O, was found. This behavior can be attributed to a dissimilar geometric arrangement and hydrogen bonding profile between both molecules in this area. Collectively, these findings suggest mechanistic heterogeneity in H2O and H2O2 permeation through PIPs.

Asunto(s)

Acuaporinas , Peróxido de Hidrógeno , Simulación de Dinámica Molecular , Proteínas de Plantas , Agua , Peróxido de Hidrógeno/metabolismo , Acuaporinas/metabolismo , Acuaporinas/química , Acuaporinas/genética , Agua/metabolismo , Agua/química , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Medicago truncatula/metabolismo , Medicago truncatula/genética , Membrana Celular/metabolismo , Enlace de Hidrógeno

RESUMEN

Plant peptidase inhibitors play crucial roles in plant defence mechanisms and physiological processes. In this study, we isolated and characterised a Kunitz trypsin inhibitor from Enterolobium gummiferum seeds named EgPI (E. gummiferum peptidase inhibitor). The purification process involved two chromatography steps using size exclusion and hydrophobic resins, resulting in high purity and yield. EgPI appeared as a single band of ~20 kDa in SDS-PAGE. Under reducing conditions, the inhibitor exhibited two polypeptide chains, with 15 and 5 kDa. Functional characterisation revealed that EgPI displayed an inhibition stoichiometry of 1:1 against trypsin, with a dissociation constant of 8.4 × 10-9 mol·L-1. The amino-terminal sequencing of EgPI revealed the homology with Kunitz inhibitors. Circular dichroism analysis provided insights into the secondary structure of EgPI, which displayed the signature typical of Kunitz inhibitors. Stability studies demonstrated that EgPI maintained the secondary structure necessary to exhibit its inhibitory activity up to 70 °C and over a pH range from 2 to 8. Microbiological screening revealed that EgPI has antibiofilm properties against pathogenic yeasts at 1.125 µmol·L-1, and EgPI reduced C. albicans biofilm formation by 82.7%. The high affinity of EgPI for trypsin suggests potential applications in various fields. Furthermore, its antibiofilm properties recommended its usefulness in agriculture and antimicrobial therapy research, highlighting the practical implications of our research.

Asunto(s)

Biopelículas , Fabaceae , Proteínas de Plantas , Semillas , Inhibidores de Tripsina , Semillas/química , Biopelículas/efectos de los fármacos , Fabaceae/química , Inhibidores de Tripsina/farmacología , Inhibidores de Tripsina/química , Inhibidores de Tripsina/aislamiento & purificación , Proteínas de Plantas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Candida albicans/efectos de los fármacos , Antifúngicos/farmacología , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Secuencia de Aminoácidos , Péptidos

RESUMEN

Alternative protein sources have been required to meet the significant plant protein demand. Agro-industrial by-products such as leaves have considerable potential as a source of macromolecules once they are mostly discarded as waste. The current study evaluated dried cassava leaves as a protein source. First, alkaline extraction parameters (solid-liquid ratio, pH, and temperature) were optimized and the run that result in the highest protein yield were acidified at pH 2.5 or 4. The influence of carbohydrate solubilized on protein precipitation was also evaluated by removing it via alcoholic extraction prior to precipitation. The experimental design showed that high pH and temperature conditions associated with a low solid-liquid ratio led to increased protein yields. The presence of carbohydrates in the supernatant significantly influenced protein precipitation. The protein concentrate had around 17.51% protein when it was obtained from a supernatant with carbohydrates, while protein content increased to 26.88% when it was obtained from carbohydrate-free supernatant. The precipitation pH also influenced protein content, whereas protein content significantly decreased when pH increased from 2.5 to 4. The natural interaction between carbohydrates and proteins from cassava leaves positively influenced the emulsion stability index and the foaming capacity and stability. Thus, the presented results bring insights into challenges in extracting and precipitation proteins from agro-industrial by-products.

Asunto(s)

Precipitación Química , Manihot , Hojas de la Planta , Proteínas de Plantas , Temperatura , Manihot/química , Hojas de la Planta/química , Concentración de Iones de Hidrógeno , Proteínas de Plantas/aislamiento & purificación , Proteínas de Plantas/química , Emulsiones/química

RESUMEN

Legumes are abundant sources of proteins, and white common bean proteins play an important role in air-water interface properties. This study aims to investigate the technical-functional properties of white common bean protein isolate (BPI) as a function of pH, protein concentration, and guar gum (GG) presence. BPI physicochemical properties were analyzed in terms of solubility, zeta potential, and mean particle diameter at pH ranging from 2 to 9, in addition to water-holding capacity (WHC), oil-holding capacity (OHC), and thermogravimetric analysis. Protein dispersions were evaluated in terms of dynamic, interfacial, and foam-forming properties. BPI showed higher solubility (>80 %) at pH 2 and above 7. Zeta potential and mean diameter ranged from 15.43 to -34.08 mV and from 129.55 to 139.90 nm, respectively. BPI exhibited WHC and OHC of 1.37 and 4.97 g/g, respectively. Thermograms indicated decomposition temperature (295.81 °C) and mass loss (64.73 %). Flow curves indicated pseudoplastic behavior, with higher η100 values observed in treatments containing guar gum. The behavior was predominantly viscous (tg δ > 1) at lower frequencies, at all pH levels, shifting to predominantly elastic at higher frequencies. Equilibrium surface tension (γeq) ranged from 43.87 to 41.95 mN.m-1 and did not decrease with increasing protein concentration under all pH conditions. All treatments exhibited ϕ < 15°, indicating predominantly elastic surface films. Foaming properties were influenced by higher protein concentration and guar gum addition, and the potential formation of protein-polysaccharide complexes favored the kinetic stability of the system.

Asunto(s)

Galactanos , Mananos , Phaseolus , Gomas de Plantas , Proteínas de Plantas , Solubilidad , Propiedades de Superficie , Gomas de Plantas/química , Galactanos/química , Mananos/química , Concentración de Iones de Hidrógeno , Proteínas de Plantas/química , Phaseolus/química , Tamaño de la Partícula , Agua/química

RESUMEN

With the emergence of multidrug-resistant microorganisms, microbial agents have become a serious global threat, affecting human health and various plants. Therefore, new therapeutic alternatives, such as chitin-binding proteins, are necessary. Chitin is an essential component of the fungal cell wall, and chitin-binding proteins exhibit antifungal activity. In the present study, chitin-binding peptides isolated from Capsicum chinense seeds were characterized and evaluated for their in vitro antimicrobial effect against the growth of Candida and Fusarium fungi. Proteins were extracted from the seeds and subsequently the chitin-binding proteins were separated by chitin affinity chromatography. After chromatography, two fractions, Cc-F1 (not retained on the column) and Cc-F2 (retained on the column), were obtained. Electrophoresis revealed major protein bands between 6.5 and 26.6 kDa for Cc-F1 and only a ~ 6.5 kDa protein band for Cc-F2, which was subsequently subjected to mass spectrometry. The protein showed similarity with hevein-like and endochitinase and was then named Cc-Hev. Data are available via ProteomeXchange with identifier PXD054607. Next, we predicted the three-dimensional structure of the peptides and performed a peptide docking with (NAG)3. Subsequently, growth inhibition assays were performed to evaluate the ability of the peptides to inhibit microorganism growth. Cc-Hev inhibited the growth of C. albicans (up to 75% inhibition) and C. tropicalis (100% inhibition) and induced a 65% decrease in cell viability for C. albicans and 100% for C. tropicalis. Based on these results, new techniques to combat fungal diseases could be developed through biotechnological applications; therefore, further studies are needed.

Asunto(s)

Antifúngicos , Candida , Capsicum , Quitina , Quitinasas , Fusarium , Semillas , Semillas/química , Antifúngicos/farmacología , Antifúngicos/aislamiento & purificación , Antifúngicos/química , Antifúngicos/metabolismo , Quitina/metabolismo , Quitina/farmacología , Fusarium/efectos de los fármacos , Quitinasas/farmacología , Quitinasas/metabolismo , Quitinasas/química , Quitinasas/aislamiento & purificación , Candida/efectos de los fármacos , Candida/enzimología , Lectinas de Plantas/farmacología , Lectinas de Plantas/química , Lectinas de Plantas/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Péptidos/farmacología , Péptidos/química , Péptidos/aislamiento & purificación , Péptidos/metabolismo , Simulación del Acoplamiento Molecular , Proteínas de Plantas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Proteínas de Plantas/metabolismo , Péptidos Catiónicos Antimicrobianos

RESUMEN

Cancer prevails as one of the major health concerns worldwide due to the consistent rise in incidence and lack of effective therapies. Previous studies identified the peptides KLKKNL, MLKSKR, and KKYRVF from Salvia hispanica seeds and stated their selective anticancer activity. Thus, this study aimed to determine the cell death pathway induced by these peptides on five cancer cell lines (MCF-7, Caco2, HepG2, DU145, and HeLa). Based on the results of this work, it is possible to suggest that KLKKNL primarily induces selective cancer cell death through the apoptotic pathway in the Caco2 and HeLa lines. On the other hand, the peptide KKYRVF reported the highest statistical (p < 0.05) selective cytotoxic effect on the MCF-7, Caco2, HepG2, and DU145 cancer cell lines by induction of the necrotic pathway. These findings offer some understanding of the selective anticancer effect of KLKKNL, MLKSKR, and KKYRVF.

Asunto(s)

Apoptosis , Péptidos , Salvia , Semillas , Humanos , Semillas/química , Péptidos/farmacología , Péptidos/química , Apoptosis/efectos de los fármacos , Salvia/química , Línea Celular Tumoral , Extractos Vegetales/farmacología , Extractos Vegetales/química , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Proliferación Celular/efectos de los fármacos , Antineoplásicos Fitogénicos/farmacología , Antineoplásicos Fitogénicos/química , Supervivencia Celular/efectos de los fármacos , Proteínas de Plantas/farmacología , Proteínas de Plantas/química

RESUMEN

Protein hydrolysates with antioxidant potential have been reported to act as adjuvants in preventing and treating type-2 diabetes (T2D). This work investigated the biochemical, antidiabetic, antioxidant potential, and physicochemical properties of chia meal protein hydrolysate (CMPH). Bands smaller than 14 kDa were observed in the electrophoretic profile. The predominant amino acids were hydrophobic and aromatic. CMPH had the potential to inhibit α-amylase (IC50: 1.76 ± 0.13 mg/mL), α-glucosidase (IC50: 0.42 ± 0.13 mg/mL), and DPP-IV (IC50: 0.46 ± 0.14 mg/mL). Antioxidant activity for ABTS (IC50: 0.236 mg/mL), DPPH (8.83 ± 0.52%), and ORAC (IC25: 0.115 mg/mL). Against chia meal protein isolate (CMPI), CMPH has a broad solubility (pH 2-12.46). Particle size (624.5 ± 247.3 nm), low PDI (0.22 ± 0.06), ζ-potential (-31.1 ± 2.5 mV), and surface hydrophobicity (11,183.33 ± 2024.11) and the intrinsic fluorescence peak of CMPH was lower than that of CMPI. CMPH represents an alternative to add value to the agri-food co-product of the chia seed oil industry, generating food ingredients with outstanding antidiabetic and antioxidant potential.

Asunto(s)

Antioxidantes , Hipoglucemiantes , Hidrolisados de Proteína , Salvia hispanica , alfa-Amilasas , Hipoglucemiantes/química , Antioxidantes/química , Hidrolisados de Proteína/química , alfa-Amilasas/química , Salvia hispanica/química , alfa-Glucosidasas/química , alfa-Glucosidasas/metabolismo , Humanos , Inhibidores de Glicósido Hidrolasas/química , Inhibidores de Glicósido Hidrolasas/farmacología , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Proteínas de Plantas/química , Interacciones Hidrofóbicas e Hidrofílicas , Salvia/química

RESUMEN

Faced with the emergence of multiresistant microorganisms that affect human health, microbial agents have become a serious global threat, affecting human health and plant crops. Antimicrobial peptides have attracted significant attention in research for the development of new microbial control agents. This work's goal was the structural characterization and analysis of antifungal activity of chitin-binding peptides from Capsicum baccatum and Capsicum frutescens seeds on the growth of Candida and Fusarium species. Proteins were initially submitted to extraction in phosphate buffer pH 5.4 and subjected to chitin column chromatography. Posteriorly, two fractions were obtained for each species, Cb-F1 and Cf-F1 and Cb-F2 and Cf-F2, respectively. The Cb-F1 (C. baccatum) and Cf-F1 (C. frutescens) fractions did not bind to the chitin column. The electrophoresis results obtained after chromatography showed two major protein bands between 3.4 and 14.2 kDa for Cb-F2. For Cf-F2, three major bands were identified between 6.5 and 14.2 kDa. One band from each species was subjected to mass spectrometry, and both bands showed similarity to nonspecific lipid transfer protein. Candida albicans and Candida tropicalis had their growth inhibited by Cb-F2. Cf-F2 inhibited the development of C. albicans but did not inhibit the growth of C. tropicalis. Both fractions were unable to inhibit the growth of Fusarium species. The toxicity of the fractions was tested in vivo on Galleria mellonella larvae, and both showed a low toxicity rate at high concentrations. As a result, the fractions have enormous promise for the creation of novel antifungal compounds.

Asunto(s)

Antifúngicos , Candida , Quitina , Fusarium , Simulación del Acoplamiento Molecular , Antifúngicos/farmacología , Antifúngicos/química , Antifúngicos/metabolismo , Quitina/química , Quitina/metabolismo , Fusarium/efectos de los fármacos , Candida/efectos de los fármacos , Proteínas Portadoras/química , Proteínas Portadoras/metabolismo , Animales , Capsicum/química , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proteínas de Plantas/farmacología , Pruebas de Sensibilidad Microbiana , Unión Proteica , Conformación Proteica

RESUMEN

The Jatropha curcas cake, a protein-rich by-product of biofuel production, was the subject of our study. We identified and quantified the ACE inhibitory, antioxidant, and antidiabetic activities of bioactive peptides from a Jatropha curcas L. var Sevangel protein isolate. The protein isolate (20.44% recovered dry matter, 38.75% protein content, and 34.98% protein yield) was subjected to two enzyme systems for hydrolysis: alcalase (PEJA) and flavourzyme (PEJF), recording every 2 h until 8 h had passed. The highest proteolytic capacity in PEJA was reached at 2 h (4041.38 ± 50.89), while in PEJF, it was reached at 6 h (3435.16 ± 59.31). Gel electrophoresis of the PEJA and PEJF samples showed bands corresponding to peptides smaller than 10 kDa in both systems studied. The highest values for the antioxidant capacity (DPPH) were obtained at 4 h for PEJA (56.17 ± 1.14), while they were obtained at 6 h for PEJF (26.64 ± 0.52). The highest values for the antihypertensive capacity were recorded at 6 h (86.46 ± 1.85) in PEJF. The highest antidiabetic capacity obtained for PEJA and PEJF was observed at 6 h, 68.86 ± 8.27 and 52.75 ± 2.23, respectively. This is the first report of their antidiabetic activity. Notably, alcalase hydrolysate outperformed flavourzyme hydrolysate and the cereals reported in other studies, confirming its better multi-bioactivity.

Asunto(s)

Inhibidores de la Enzima Convertidora de Angiotensina , Antioxidantes , Hipoglucemiantes , Jatropha , Proteínas de Plantas , Jatropha/química , Hidrólisis , Antioxidantes/química , Antioxidantes/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/química , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Subtilisinas/metabolismo , Subtilisinas/química , Endopeptidasas

RESUMEN

Tribolium castaneum, also known as the red flour beetle, is a polyphagous pest that seriously damages agricultural products, including stored and processed grains. Researchers have aimed to discover alternative pest control mechanisms that are less harmful to the ecosystem than those currently used. We conduct the purification and characterization of a protease inhibitor from C. plumieri seeds and an in vitro evaluation of its insecticidal potential against the insect pest T. castaneum. The trypsin inhibitor was isolated from C. plumieri seeds in a single-step DEAE-Sepharose column chromatography and had a molecular mass of 50 kDA. When analyzed for interaction with different proteolytic enzymes, the inhibitor exhibited specificity against trypsin and no activity against other serine proteases such as chymotrypsin and elastase-2. The isolated inhibitor was able to inhibit digestive enzymes of T. castaneum from extracts of the intestine of this insect. Therefore, we conclude that the new protease inhibitor, specific in tryptic inhibition, of protein nature from the seeds of C. plumieri was effective in inhibiting the digestive enzymes of T. castaneum and is a promising candidate in the ecological control of pests.

Asunto(s)

Tribolium , Inhibidores de Tripsina , Animales , Inhibidores de Tripsina/farmacología , Inhibidores de Tripsina/química , Inhibidores de Tripsina/aislamiento & purificación , Tribolium/enzimología , Tribolium/efectos de los fármacos , Proteínas de Insectos/química , Proteínas de Insectos/aislamiento & purificación , Proteínas de Insectos/antagonistas & inhibidores , Semillas/química , Insecticidas/farmacología , Insecticidas/química , Insecticidas/aislamiento & purificación , Proteínas de Plantas/farmacología , Proteínas de Plantas/aislamiento & purificación , Proteínas de Plantas/química

RESUMEN

RAMOSA1 (RA1) is a Cys2-His2-type (C2H2) zinc finger transcription factor that controls plant meristem fate and identity and has played an important role in maize domestication. Despite its importance, the origin of RA1 is unknown, and the evolution in plants is only partially understood. In this paper, we present a well-resolved phylogeny based on 73 amino acid sequences from 48 embryophyte species. The recovered tree topology indicates that, during grass evolution, RA1 arose from two consecutive SUPERMAN duplications, resulting in three distinct grass sequence lineages: RA1-like A, RA1-like B, and RA1; however, most of these copies have unknown functions. Our findings indicate that RA1 and RA1-like play roles in the nucleus despite lacking a traditional nuclear localization signal. Here, we report that copies diversified their coding region and, with it, their protein structure, suggesting different patterns of DNA binding and protein-protein interaction. In addition, each of the retained copies diversified regulatory elements along their promoter regions, indicating differences in their upstream regulation. Taken together, the evidence indicates that the RA1 and RA1-like gene families in grasses underwent subfunctionalization and neofunctionalization enabled by gene duplication.

Asunto(s)

Evolución Molecular , Filogenia , Proteínas de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Embryophyta/genética , Embryophyta/metabolismo , Secuencia de Aminoácidos

RESUMEN

The obtained seeds from fruit processing are considered by-products containing proteins that could be utilized as ingredients in food manufacturing. However, in the specific case of soursop seeds, their usage for the preparation of protein isolates is limited. In this investigation a protein isolate from soursop seeds (SSPI) was obtained by alkaline extraction and isoelectric precipitation methods. The SSPI was sonicated at 200, 400 and 600 W during 15 and 30 min and its effect on the physicochemical, functional, biochemical, and structural properties was evaluated. Ultrasound increased (p < 0.05) up to 5 % protein content, 261 % protein solubility, 60.7 % foaming capacity, 30.2 % foaming stability, 86 % emulsifying activity index, 4.1 % emulsifying stability index, 85.4 % in vitro protein digestibility, 423.4 % albumin content, 83 % total sulfhydryl content, 316 % free sulfhydryl content, 236 % α-helix, 46 % ß-sheet, and 43 % ß-turn of SSPI, in comparison with the control treatment without ultrasound. Furthermore, ultrasound decreased (p < 0.05) up to 50 % particle size, 37 % molecular flexibility, 68 % surface hydrophobicity, 41 % intrinsic florescence spectrum, and 60 % random coil content. Scanning electron microscopy analysis revealed smooth structures of the SSPI with molecular weights ranging from 12 kDa to 65 kDa. The increase of albumins content in the SSPI by ultrasound was highly correlated (r = 0.962; p < 0.01) with the protein solubility. Improving the physicochemical, functional, biochemical and structural properties of SSPI by ultrasound could contribute to its utilization as ingredient in food industry.

Asunto(s)

Annona , Proteínas de Plantas , Semillas , Solubilidad , Semillas/química , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Annona/química , Ondas Ultrasónicas , Fenómenos Químicos , Sonicación

RESUMEN

Salinity in plants generates an osmotic and ionic imbalance inside cells that compromises the viability of the plant. Rab GTPases, the largest family within the small GTPase superfamily, play pivotal roles as regulators of vesicular trafficking in plants, including the economically important and globally cultivated tomato (Solanum lycopersicum). Despite their significance, the specific involvement of these small GTPases in tomato vesicular trafficking and their role under saline stress remains poorly understood. In this work, we identified and classified 54 genes encoding Rab GTPases in cultivated tomato, elucidating their genomic distribution and structural characteristics. We conducted an analysis of duplication events within the S. lycopersicum genome, as well as an examination of gene structure and conserved motifs. In addition, we investigated the transcriptional profiles for these Rab GTPases in various tissues of cultivated and wild tomato species using microarray-based analysis. The results showed predominantly low expression in most of the genes in both leaves and vegetative meristem, contrasting with notably high expression levels observed in seedling roots. Also, a greater increase in gene expression in shoots from salt-tolerant wild tomato species was observed under normal conditions when comparing Solanum habrochaites, Solanum pennellii, and Solanum pimpinellifolium with S. lycopersicum. Furthermore, an expression analysis of Rab GTPases from Solanum chilense in leaves and roots under salt stress treatment were also carried out for their characterization. These findings revealed that specific Rab GTPases from the endocytic pathway and the trans-Golgi network (TGN) showed higher induction in plants exposed to saline stress conditions. Likewise, disparities in gene expression were observed both among members of the same Rab GTPase subfamily and between different subfamilies. Overall, this work emphasizes the high degree of conservation of Rab GTPases, their high functional diversification in higher plants, and the essential role in mediating salt stress tolerance and suggests their potential for further exploration of vesicular trafficking mechanisms in response to abiotic stress conditions.

Asunto(s)

Proteínas de Plantas , Solanum lycopersicum , Proteínas de Unión al GTP rab , Solanum lycopersicum/enzimología , Solanum lycopersicum/genética , Proteínas de Unión al GTP rab/química , Proteínas de Unión al GTP rab/genética , Proteínas de Unión al GTP rab/metabolismo , Perfilación de la Expresión Génica , Filogenia , Duplicación de Gen , Intrones , Exones , Secuencias de Aminoácidos , Vesículas Transportadoras/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo

RESUMEN

Due to allergenic concerns, only pea, potato, and wheat proteins have been approved as alternatives for replacing animal-based fining agents in wines. In pursuit of other substitutes, this work aimed to determine the fining ability of amaranth (Amaranthus caudatus L.) proteins (AP) in red wine, compared to quinoa (Chenopodium quinoa Willd.) (QP) and a commercial pea protein. Phenolic and volatile composition, as well as color characteristics, were analyzed. AP was as effective as QP at decreasing condensed tannins, with AP at 50 g/hL being the most effective treatment (25.6% reduction). QP and AP produced a minor or no statistical change in the total anthocyanins and wine color intensity. They reduced the total ester concentration, but the total alcohols remained unchanged. The outcomes of AP and QP were similar, and sometimes better than the pea proteins, thus suggesting that they could be promising options for the development of novel fining agents.

Asunto(s)

Amaranthus , Chenopodium quinoa , Proteínas de Plantas , Vino , Amaranthus/química , Chenopodium quinoa/química , Proteínas de Plantas/química , Proteínas de Plantas/análisis , Vino/análisis , Fenoles/química , Fenoles/análisis , Extractos Vegetales/química , Color

RESUMEN

In the present study, we identified and characterized two defensin-like peptides in an antifungal fraction obtained from Capsicum chinense pepper fruits and inhibited the growth of Colletotrichum scovillei, which causes anthracnose. AMPs were extracted from the pericarp of C. chinense peppers and subjected to ion exchange, molecular exclusion, and reversed-phase in a high-performance liquid chromatography system. We investigated the endogenous increase in reactive oxygen species (ROS), the loss of mitochondrial functioning, and the ultrastructure of hyphae. The peptides obtained from the G3 fraction through molecular exclusion chromatography were subsequently fractionated using reverse-phase chromatography, resulting in the isolation of fractions F1, F2, F3, F4, and F5. The F1-Fraction suppressed C. scovillei growth by 90, 70.4, and 44% at 100, 50, and 25 µg mL-1, respectively. At 24 h, the IC50 and minimum inhibitory concentration were 21.5 µg mL-1 and 200 µg mL-1, respectively. We found an increase in ROS, which may have resulted in an oxidative burst, loss of mitochondrial functioning, and cytoplasm retraction, as well as an increase in autophagic vacuoles. MS/MS analysis of the F1-Fraction indicated the presence of two defensin-like proteins, and we were able to identify the expression of three defensin sequences in our C. chinense fruit extract. The F1-Fraction was also found to inhibit the activity of insect α-amylases. In summary, the F1-Fraction of C. chinense exhibits antifungal activity against a major pepper pathogen that causes anthracnose. These defensin-like compounds are promising prospects for further research into antifungal and insecticide biotechnology applications. © 2024 Society of Chemical Industry.

Asunto(s)

Capsicum , Colletotrichum , Defensinas , Mitocondrias , Especies Reactivas de Oxígeno , Colletotrichum/efectos de los fármacos , Colletotrichum/crecimiento & desarrollo , Capsicum/microbiología , Especies Reactivas de Oxígeno/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Defensinas/farmacología , Defensinas/química , Antifúngicos/farmacología , Antifúngicos/química , Proteínas de Plantas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Frutas/microbiología

RESUMEN

Alterations in cell wall composition imply in new structural and functional traits in gall developmental sites, even when the inducer is a sucking exophytophagous insect with strict feeding sites as the aphid associated to Malus domestica Borkh. This host plant is an economically important, fruit-bearing species, susceptible to gall induction by the sucking aphid Eriosoma lanigerum Hausmann, 1802. Herein, the immunocytochemical detection of arabinogalactan-proteins (AGPs), pectins, and hemicelluloses using monoclonal antibodies was performed in samples of non-galled roots and stems, and of root and stem galls on M. domestica. The dynamics of these cell wall components was discussed under the structural and functional traits of the galls proximal, median, and distal regions, according to the proximity of E. lanigerum colony feeding site. In the proximal region, the epitopes of AGPs and homogalacturonans (HGs) are related to cell growth and divisions, which result in the overproduction of parenchyma cells both in root and stem galls. In the proximal and median regions, the co-occurrence of HGs and arabinans in the cell walls of parenchyma and secondary tissues favors the nutrient flow and water-holding capacity, while the xylogalacturonans and hemicelluloses may function as additional carbohydrate resources to E. lanigerum. The immunocytochemical profile of the cell walls support the feeding activity of E. lanigerum mainly in the gall proximal region. The similarity of the cell wall components of the gall distal region and the non-galled portions, both in roots and stems, relates to the decrease of the cecidogenetic field the more distant the E. lanigerum colony is.

Asunto(s)

Pared Celular , Inmunohistoquímica , Malus , Raíces de Plantas , Tallos de la Planta , Tumores de Planta , Pared Celular/química , Pared Celular/metabolismo , Animales , Raíces de Plantas/parasitología , Tallos de la Planta/química , Tumores de Planta/parasitología , Mucoproteínas/metabolismo , Áfidos/fisiología , Polisacáridos/metabolismo , Polisacáridos/química , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Pectinas/metabolismo

RESUMEN

A novel trypsin inhibitor from Cajanus cajan (TIC) fresh leaves was partially purified by affinity chromatography. SDS-PAGE revealed one band with about 15 kDa with expressive trypsin inhibitor activity by zymography. TIC showed high affinity for trypsin (Ki = 1.617 µM) and was a competitive inhibitor for this serine protease. TIC activity was maintained after 24 h of treatment at 70 °C, after 1 h treatments with different pH values, and ß-mercaptoethanol increasing concentrations, and demonstrated expressive structural stability. However, the activity of TIC was affected in the presence of oxidizing agents. In order to study the effect of TIC on secreted serine proteases, as well as on the cell culture growth curve, SK-MEL-28 metastatic human melanoma cell line and CaCo-2 colon adenocarcinoma was grown in supplemented DMEM, and the extracellular fractions were submitted salting out and affinity chromatography to obtain new secreted serine proteases. TIC inhibited almost completely, 96 to 89%, the activity of these serine proteases and reduced the melanoma and colon adenocarcinoma cells growth of 48 and 77% respectively. Besides, it is the first time that a trypsin inhibitor was isolated and characterized from C. cajan leaves and cancer serine proteases were isolated and partial characterized from SK-MEL-28 and CaCo-2 cancer cell lines. Furthermore, TIC shown to be potent inhibitor of tumor protease affecting cell growth, and can be one potential drug candidate to be employed in chemotherapy of melanoma and colon adenocarcinoma.

Asunto(s)

Cajanus , Hojas de la Planta , Humanos , Cajanus/química , Hojas de la Planta/química , Células CACO-2 , Proliferación Celular/efectos de los fármacos , Línea Celular Tumoral , Inhibidores de Tripsina/farmacología , Inhibidores de Tripsina/química , Inhibidores de Tripsina/aislamiento & purificación , Proteínas de Plantas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Serina Proteasas/química , Serina Proteasas/aislamiento & purificación , Serina Proteasas/metabolismo