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1.
Metabolites ; 13(3)2023 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-36984833

RESUMEN

This investigation aimed to determine the effect of high-power ultrasonic pulses on the antioxidant, antibacterial, and antiproliferative activity of sorghum (Sorghum bicolor) lignin. A lignin yield of 7.35% was obtained using the organosolv method. Additionally, the best conditions of the ultrasonic pulses were optimized to obtain a more significant increase in antioxidant capacity, resulting in 10 min for all treatments, with amplitudes of 20% for DPPH and FRAP, 18% for ABTS, and 14% for total phenols. The effect of ultrasonic pulses was mainly observed with FRAP (1694.88 µmol TE/g), indicating that the main antioxidant mechanism of lignin is through electron transport. Sorghum lignin with and without ultrasonic pulses showed high percentages of hemolysis inhibition (>80%) at concentrations of 0.003 to 0.33 mg/mL. The AB blood group and, in general, all Rh- groups are the most susceptible to hemolysis. Lignin showed high anti-inflammatory potential due to heat and hypotonicity (>82%). A higher antimicrobial activity of lignin on Escherichia coli bacteria was observed. The lignins evaluated without sonication and sonication presented higher activity in the cell line PC-3. No effect was observed on the lignin structure with the FT-IR technique between sonication and non-sonication; however, the organosolv method helped extract pure lignin according to HPLC.

2.
Metabolites ; 12(12)2022 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-36557241

RESUMEN

Previous studies have reported that different blood groups are associated with the risk of chronic degenerative diseases that mainly involve inflammation and neoplastic processes. We investigate the relationship between blood groups and the erythroprotective effect of extracts from Navicula incerta against oxidative damage as a proposal to develop drugs designed for people with a specific blood type related to chronic pathology. The study was carried out through the elucidation of the erythroprotective potential, anti-inflammatory and antiproliferative activity of Navicula incerta. Research suggests that the presence or absence of certain blood groups increases or decreases the abilities of certain phytochemicals to inhibit oxidative stress, which is related to the systemic inflammatory response involved in the development of different types of cancer. The pigment-rich extracts from Navicula incerta inhibit ROO•- induced oxidative stress in human erythrocytes on the A RhD+ve antigen without compromising the structure of the cell membrane. This result is very important, since the A antigen is related to the susceptibility of contracting prostate cancer. Similarly, it was possible to inhibit the proliferation of cervical (HeLa) and prostate (PC-3) carcinoma. The combinatorial analysis of different biological activities can help design phytochemicals as new candidates for preventive drugs treating the chronic degenerative diseases associated with a specific blood group.

3.
Nanomaterials (Basel) ; 12(16)2022 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-36014684

RESUMEN

The objective of this paper was to report the effect of ionic liquids (ILs) in the elaboration of nanofibers of cellulose bagasse from Agave tequilana Weber var. azul by the electrospinning method. The ILs used were 1-butyl-3-methylimidazolium chloride (BMIMCl), and DMSO was added as co-solvent. To observe the effect of ILs, this solvent was compared with the organic solvent TriFluorAcetic acid (TFA). The nanofibers were characterized by transmission electron microscopy (TEM), X-ray, Fourier transform-infrared using attenuated total reflection (FTIR-ATR) spectroscopy, and thermogravimetric analysis (TGA). TEM showed different diameters (ranging from 35 to 76 nm) of cellulose nanofibers with ILs (CN ILs). According to X-ray diffraction, a notable decrease of the crystalline structure of cellulose treated with ILs was observed, while FTIR-ATR showed two bands that exhibit the physical interaction between cellulose nanofibers and ILs. TGA revealed that CN ILs exhibit enhanced thermal properties due to low or null cellulose crystallinity. CN ILs showed better characteristics in all analyses than nanofibers elaborated with TFA organic solvent. Therefore, CN ILs provide new alternatives for cellulose bagasse. Due to their small particle size, CN ILs could have several applications, including in food, pharmaceutical, textile, and material areas, among others.

4.
Molecules ; 26(9)2021 May 08.
Artículo en Inglés | MEDLINE | ID: mdl-34066776

RESUMEN

The aim of this work was to monitor the quality, antioxidant capacity and digestibility of chickpea exposed to different modified atmospheres. Chickpea quality (proximal analysis, color, texture, and water absorption) and the antioxidant capacity of free, conjugated, and bound phenol fractions obtained from raw and cooked chickpea, were determined. Cooked chickpea was exposed to N2 and CO2 atmospheres for 0, 25, and 50 days, and the antioxidant capacity was analyzed by DPPH (2,2'-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis-[3ethylbenzothiazoline-6-sulfonic acid]), and total phenols. After in vitro digestion, the antioxidant capacity was measured by DPPH, ABTS, FRAP (ferric reducing antioxidant power), and AAPH (2,2'-Azobis [2-methylpropionamidine]). Additionally, quantification of total phenols, and UPLC-MS profile were determined. The results indicated that this grain contain high quality and high protein (18.38%). Bound phenolic compounds showed the highest amount (105.6 mg GAE/100 g) and the highest antioxidant capacity in all techniques. Cooked chickpeas maintained their quality and antioxidant capacity during 50 days of storage at 4 and -20 °C under a nitrogen atmosphere. Free and conjugated phenolic compounds could be hydrolyzed by digestive enzymes, increasing their bioaccessibility and their antioxidant capacity during each step of digestion. The majority compound in all samples was enterodiol, prevailing the flavonoid type in the rest of the identified compounds. Chickpea contains biological interest compounds with antioxidant potential suggesting that this legume can be exploited for various technologies.


Asunto(s)
Antioxidantes/química , Atmósfera/química , Dióxido de Carbono , Cicer/química , Grano Comestible/química , Nitrógeno , Cromatografía Liquida , Digestión , Flavonoides/química , Lignanos/química , Espectrometría de Masas , Fenoles/química , Proteínas de Plantas/química
5.
Saudi J Biol Sci ; 28(2): 1401-1416, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33613070

RESUMEN

Navicula incerta is a marine microalga distributed in Baja California, México, commonly used in aquaculture nutrition, and has been extended to human food, biomedical, and pharmaceutical industries due to its high biological activity. Therefore, the study aimed to optimize culture conditions to produce antioxidant pigments. A central composite experimental design and response surface methodology (RSM) was employed to analyze the best culture conditions. The medium (nitrogen-deficient concentrations), salinity (PSU = Practical Salinity Unity [g/kg]), age of culture (days), and solvent extraction (ethanol, methanol, and acetone) were the factors used for the experiment. Chlorophyll a (Chl a) and total carotenoids (T-Car), determined spectroscopically, were used as the response variables. The antioxidant capacity was evaluated by DPPH• and ABTS•+ radical inhibition, FRAP, and anti-hemolytic activity. According to the overlay plots, the optimum growth conditions for Chl a and T-Car production were the following conditions: medium = 0.44 mol·L-1 of NaNO3, salinity = 40 PSU, age of culture: 3.5 days, and solvent = methanol. The pigment extracts obtained in these optimized conditions had high antioxidant activity in ABTS•+ (86.2-92.1% of inhibition) and anti-hemolytic activity (81.8-96.7% of hemolysis inhibition). Low inhibition (33-35%) was observed in DPPH•. The highest value of FRAP (766.03 ± 16.62 µmol TE/g) was observed in the acetonic extract. The results demonstrated that RSM could obtain an extract with high antioxidant capacity with potential applications in the biomedical and pharmaceutical industry, which encourages the use of natural resources for chemoprevention of chronic-degenerative pathologies.

6.
Artículo en Inglés | MEDLINE | ID: mdl-33130328

RESUMEN

Shrimp are increasingly exposed to warmer temperatures and lower oxygen concentrations in their habitat due to climate change. These conditions may lead to oxidative stress and apoptosis. We studied the effects of high temperature, hypoxia, reoxygenation, and the combination of these factors on lipid peroxidation, protein carbonylation, and caspase-3 activity in gills of white shrimp Litopenaeus vannamei. Silencing of mitochondrial manganese superoxide dismutase (mMnSOD) was used to determine the role of this enzyme in response to the abiotic stressors described above, to avoid oxidative damage and apoptosis. In addition, mMnSOD gene expression and mitochondrial SOD activity were evaluated to determine the efficiency of silencing this enzyme. The results showed that there was no effect of the abiotic stress conditions on the thiobarbituric acid reactive substances (TBARS), but protein carbonylation increased in all the oxidative stress treatments and caspase-3 activity decreased in hypoxia at 28 °C. On the other hand, mMnSOD-silenced shrimp experienced higher oxidative stress, since TBARS, carbonylated proteins and caspase-3 activity increased in some silenced treatments. Unexpectedly, mitochondrial SOD activity increased in some of the silenced treatments as well. Altogether, these results suggest that mMnSOD has a key role in shrimp for the prevention of oxidative damage development and induction of apoptosis in response to hypoxia, reoxygenation, high temperature, and their interactions, as conditions derived from climate change.


Asunto(s)
Caspasa 3/metabolismo , Crustáceos/fisiología , Técnicas de Silenciamiento del Gen , Calor , Hipoxia/metabolismo , Mitocondrias/enzimología , Estrés Oxidativo/genética , Oxígeno/metabolismo , Superóxido Dismutasa/genética , Animales , Crustáceos/metabolismo , Silenciador del Gen , Superóxido Dismutasa/metabolismo
7.
Indian J Microbiol ; 60(4): 458-467, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33087995

RESUMEN

This study aimed to determine the ability of Fusarium verticillioides in developing mechanisms to counteract the antifungal effect of a fraction from Jacquinia macrocarpa plant extract (JmAF), as well as the morphological and physiological changes that occur during its exposure. The fungus was exposed to JmAF during consecutive periods. A culture sample was taken weekly to determine radial growth, spore germination and size, and fungal ß-1,3-glucanase activity. The results showed that, in the beginning, the radial growth decreased by 85.8%, and spore germination was delayed. As the exposure continued, the fungus showed a recovery, to some extent, in its original characteristics. However, the radial growth of the fungus continued to be inhibited (42.9%) throughout the experiment (7 weeks). The ß-1,3-glucanase activity also was inhibited by 36.4% during the first week of exposure to JmAF. However, the activity was recovered after 7 weeks of exposure.

8.
Int J Biol Macromol ; 164: 2701-2710, 2020 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-32827617

RESUMEN

Trypsins (E.C. 3.4.21.4) are digestive enzymes that catalyze the hydrolysis of peptide bonds containing arginine and lysine residues. Some trypsins from fish species are active at temperatures just above freezing, and for that are called cold-adapted enzymes, having many biotechnological applications. In this work, we characterized a recombinant trypsin-III from Monterey sardine (Sardinops caeruleus) and studied the role of a single residue on its cold-adapted features. The A236N mutant from sardine trypsin-III showed higher activation energy for the enzyme-catalyzed reaction, it was more active at higher temperatures, and exhibited a higher thermal stability than the wild-type enzyme, suggesting a key role of this residue. The thermodynamic activation parameters revealed an increase in the activation enthalpy for the A236N mutant, suggesting the existence of more intramolecular contacts during the activation step. Molecular models for both enzymes suggest that a hydrogen-bond involving N236 may contact the C-terminal α-helix to the vicinity of the active site, thus affecting the biochemical and thermodynamic properties of the enzyme.


Asunto(s)
Peces/metabolismo , Mutación , Tripsina/química , Tripsina/genética , Animales , Frío , Activación Enzimática , Estabilidad de Enzimas , Proteínas de Peces/química , Proteínas de Peces/genética , Peces/genética , Enlace de Hidrógeno , Modelos Moleculares , Simulación del Acoplamiento Molecular , Estructura Secundaria de Proteína
9.
Molecules ; 24(23)2019 Nov 29.
Artículo en Inglés | MEDLINE | ID: mdl-31795320

RESUMEN

Pistachio nuts are included among the foods with the highest antioxidant capacity. Stressed cultivating conditions, such as the use of regulated deficit irrigation (RDI), are expected to create a plant response that might increase the production of secondary metabolites. Fruits that are obtained under RDI treatments are commonly called hydroSOS products. The aim of this work was to study the influence of using different rootstocks (P. atlantica, P. integerrima, and P. terebinthus) and two RDI treatments on the antioxidant (ABTS, ferric reducing antioxidant power (FRAP), and DPPH), antimutagenic (Ames test), and cytotoxicity (MTT assay in five human cell lines) activities of pistachios. P. terebinthus showed the best antioxidant activity, and the RDI treatments maintained and improved the antioxidant properties of pistachios. Neither the rootstock nor the RDI had significant impact on the antimutagenic potential of pistachios. The nut extracts had no toxic effect on non-cancerous cells and the application of RDI did not reduce their cytoprotective capacity. Furthermore, neither rootstock nor RDI treatments affected the ability of the pistachio extracts of preventing the oxidative damage by H2O2. The application of RDI strategies, in addition to allowing irrigation water saving, led to obtaining pistachios with the same or even better biofunctional characteristics as compared to fully irrigated pistachios.


Asunto(s)
Antimutagênicos , Antioxidantes , Citoprotección/efectos de los fármacos , Nueces/química , Pistacia/química , Células A549 , Antimutagênicos/química , Antimutagênicos/aislamiento & purificación , Antimutagênicos/farmacología , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Humanos , Peróxido de Hidrógeno/farmacología
10.
Biomolecules ; 9(11)2019 10 31.
Artículo en Inglés | MEDLINE | ID: mdl-31683580

RESUMEN

(1) Background: Lipases and esterases are important enzymes that share the α/ß hydrolase fold. The activity and cellular localization are important characteristics to understand the role of such enzymes in an organism. (2) Methods: Bioinformatic and biochemical tools were used to describe a new α/ß hydrolase from a Litopenaeus vannamei transcriptome (LvFHS for Family Serine Hydrolase). (3) Results: The enzyme was obtained by heterologous overexpression in Escherichia coli and showed hydrolytic activity towards short-chain lipid substrates and high affinity to long-chain lipid substrates. Anti-LvFHS antibodies were produced in rabbit that immunodetected the LvFSH enzyme in several shrimp tissues. (4) Conclusions: The protein obtained and analyzed was an α/ß hydrolase with esterase and lipase-type activity towards long-chain substrates up to 12 carbons; its immunodetection in shrimp tissues suggests that it has an intracellular localization, and predicted roles in energy mobilization and signal transduction.


Asunto(s)
Hidrolasas/metabolismo , Penaeidae/enzimología , Secuencia de Aminoácidos , Animales , Hidrolasas/química , Hidrolasas/genética , Espacio Intracelular/metabolismo , Modelos Moleculares , Penaeidae/citología , Estructura Secundaria de Proteína , Serina/metabolismo , Transducción de Señal
11.
J Food Sci ; 84(10): 2883-2897, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31553062

RESUMEN

Quercetin is a hydrophobic flavonoid with high antioxidant activity. However, for biological applications, the bioavailability of quercetin is low due to physiological barriers. For this reason, an alternative is the protection of quercetin in matrices of biopolymers as zein. The objective of this work was to prepare and characterize quercetin-loaded zein nanoparticles by electrospraying and its study of in vitro bioavailability. The physicochemical parameters such as viscosity, density, and electrical conductivity of zein solutions showed a dependence of the ethanol concentration. In addition, rheological parameters demonstrated that solutions of zein in aqueous ethanol present Newtonian behavior, rebounding in the formation of nanoparticles by electrospraying, providing spherical, homogeneous, and compact morphologies, mainly at a concentration of 80% (v/v) of ethanol and of 5% (w/v) of zein. The size and shape of quercetin-loaded zein nanoparticles were studied by transmission electron microscopy (TEM), observing that it was entrapped, distributed throughout the nanoparticle of zein. Analysis by Fourier transform-infrared (FT-IR) of zein nanoparticles loaded with quercetin revealed interactions via hydrogen bonds. The efficacy of zein nanoparticles to entrap quercetin was particularly high for all quercetin concentration evaluated in this work (87.9 ± 1.5% to 93.0 ± 2.6%). The in vitro gastrointestinal release of trapped quercetin after 240 min was 79.1%, while that for free quercetin was 99.2%. The in vitro bioavailability was higher for trapped quercetin (5.9%) compared to free quercetin (1.9%), than of gastrointestinal digestion. It is concluded, that the electrospraying technique made possible the obtention of quercitin-loaded zein nanoparticles increasing their bioavailability. PRACTICAL APPLICATION: This type of nanosystems can be used in the food and pharmaceutical industry. Quercetin-loaded zein nanoparticles for its improvement compared to free quercetin can be used to decrease the prevalence of chronic degenerative diseases by increasing of the bioavailability of quercetin in the bloodstream. Other application can be as an antioxidant system in functional foods or oils to increase shelf life.


Asunto(s)
Composición de Medicamentos/métodos , Quercetina/química , Zeína/química , Antioxidantes/química , Antioxidantes/metabolismo , Disponibilidad Biológica , Biopolímeros/química , Línea Celular , Portadores de Fármacos/química , Composición de Medicamentos/instrumentación , Humanos , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Nanopartículas/química , Nanopartículas/metabolismo , Tamaño de la Partícula , Quercetina/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier
12.
J Environ Sci Health B ; 54(8): 647-654, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31146638

RESUMEN

The aim of this study was to investigate the in vitro effect of an antifungal fraction obtained from Jacquinia macrocarpa plant (JmAF) in the generation of reactive oxygen species (ROS) and the activity of the catalase (CAT) and superoxide dismutase (SOD) enzymes from Fusarium verticillioides, as well as their influence in the viability of the fungus spores. The compounds present in the JmAF were determined by gas chromatography/quadrupole time-of-flight mass spectrometry (GC/QTOF-MS). The effect of the exposition to JmAF on the generation of ROS, as well as in the CAT and SOD activities in F. verticillioides, was determined. The main compounds detected were γ-sitosterol, stephamiersine, betulinol and oleic acid. JmAF showed very high ability in inhibiting the spore viability of F. verticillioides, and their capacity to cause oxidative stress by induction of ROS production. JmAF induced the highest ROS concentration and also inhibited CAT and SOD activities. The results obtained in this study indicate that JmAF is worthy of being considered for the fight against phytopathogenic fungi.


Asunto(s)
Antifúngicos/farmacología , Catalasa/antagonistas & inhibidores , Fusarium/efectos de los fármacos , Primulaceae/química , Superóxido Dismutasa/antagonistas & inhibidores , Antifúngicos/análisis , Antifúngicos/química , Antioxidantes/metabolismo , Catalasa/metabolismo , Proteínas Fúngicas/antagonistas & inhibidores , Proteínas Fúngicas/metabolismo , Fungicidas Industriales/química , Fungicidas Industriales/farmacología , Fusarium/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismo
13.
Protein Pept Lett ; 26(7): 532-541, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30950340

RESUMEN

BACKGROUND: Cathepsin D is a lysosomal enzyme that is found in all organisms acting in protein turnover, in humans it is present in some types of carcinomas, and it has a high activity in Parkinson's disease and a low activity in Alzheimer disease. In marine organisms, most of the research has been limited to corroborate the presence of this enzyme. It is known that cathepsin D of some marine organisms has a low thermostability and that it has the ability to have activity at very acidic pH. Cathepsin D of the Jumbo squid (Dosidicus gigas) hepatopancreas was purified and partially characterized. The secondary structure of these enzymes is highly conserved so the role of temperature and pH in the secondary structure and in protein denaturation is of great importance in the study of enzymes. The secondary structure of cathepsin D from jumbo squid hepatopancreas was determined by means of circular dichroism spectroscopy. OBJECTIVE: In this article, our purpose was to determine the secondary structure of the enzyme and how it is affected by subjecting it to different temperature and pH conditions. METHODS: Circular dichroism technique was used to measure the modifications of the secondary structure of cathepsin D when subjected to different treatments. The methodology consisted in dissecting the hepatopancreas of squid and freeze drying it. Then a crude extract was prepared by mixing 1: 1 hepatopancreas with assay buffer, the purification was in two steps; the first step consisted of using an ultrafiltration membrane with a molecular cut of 50 kDa, and the second step, a pepstatin agarose resin was used to purification the enzyme. Once the enzyme was purified, the purity was corroborated with SDS PAGE electrophoresis, isoelectric point and zymogram. Circular dichroism is carried out by placing the sample with a concentration of 0.125 mg / mL in a 3 mL quartz cell. The results were obtained in mdeg (millidegrees) and transformed to mean ellipticity per residue, using 111 g/mol molecular weight/residue as average. Secondary-structure estimation from the far-UV CD spectra was calculated using K2D Dichroweb software. RESULTS: It was found that α helix decreases at temperatures above 50 °C and above pH 4. Heating the enzyme above 70°C maintains a low percentage of α helix and increases ß sheet. Far-UV CD measurements of cathepsin D showed irreversible thermal denaturation. The process was strongly dependent on the heating rate, accompanied by a process of oligomerization of the protein that appears when the sample is heated, and maintained a certain time at this temperature. An amount typically between 3 and 4% α helix of their secondary structure remains unchanged. It is consistent with an unfolding process kinetically controlled due to the presence of an irreversible reaction. The secondary structure depends on pH, and a pH above 4 causes α helix structures to be modified. CONCLUSION: In conclusion, cathepsin D from jumbo squid hepatopancreas showed retaining up to 4% α helix at 80°C. The thermal denaturation of cathepsin D at pH 3.5 is under kinetic control and follows an irreversible model.


Asunto(s)
Catepsina D/química , Decapodiformes/química , Hepatopáncreas/química , Secuencia de Aminoácidos , Animales , Concentración de Iones de Hidrógeno , Cinética , Desnaturalización Proteica , Estructura Secundaria de Proteína , Temperatura
14.
J Food Sci ; 84(4): 818-831, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30802954

RESUMEN

Currently, electrospraying is a novel process for obtaining the nanoparticles from biopolymers. Zein nanoparticles have been obtained by this method and used to protect both hydrophilic and hydrophobic antioxidant molecules from environmental factors. The objective of this work was to prepare and characterize gallic acid-loaded zein nanoparticles obtained by the electrospraying process to provide protection to gallic acid from environmental factors. Thus, it was related to the concentration of gallic acid in physicochemical and rheological properties of the electrosprayed solution, and also to equipment parameters, such as voltage, flow rate, and distance of the collector in morphology, and particle size. The physicochemical properties showed a relationship in the formation of a Taylor cone, in which at a low concentration of gallic acid (1% w/v), low viscosity (0.00464 ± 0.00001 Pa·s), and density (0.886 ± 0.00002 g/cm3 ), as well as high electrical conductivity (369 ± 4.3 µs/cm), forms a stable cone-jet mode. The rheological properties and the Power Law model of the gallic acid-zein electrosprayed solution demonstrated Newtonian behavior (n = 1). The morphology and size of the particle were dependent on the concentration of gallic acid. Electrosprayed parameters with high voltage (15 kV), low flow rate (0.1 mL/hr), and short distance (10 cm) exhibited a smaller diameter and spherical morphology. FT-IR showed interaction in the gallic acid-loaded zein nanoparticle by hydrogen bonds. Therefore, the electrospraying process is a feasible technique for obtaining gallic acid-loaded zein nanoparticles and providing potential protection to gallic acid from environmental factors.


Asunto(s)
Técnicas Electroquímicas , Ácido Gálico/química , Nanopartículas/química , Zeína/química , Antioxidantes , Biopolímeros/química , Enlace de Hidrógeno , Tamaño de la Partícula , Espectroscopía Infrarroja por Transformada de Fourier
15.
Protein Pept Lett ; 26(3): 170-175, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30338728

RESUMEN

BACKGROUND: Trypsin from fish species is considered as a cold-adapted enzyme that may find potential biotechnological applications. In this work, the recombinant expression, refolding and activation of Trypsin I (TryI) from Monterey sardine (Sardinops sagax caerulea) are reported. METHODS: TryI was overexpressed in Escherichia coli BL21 as a fusion protein of trypsinogen with thioredoxin. Refolding of trypsinogen I was achieved by dialysis of bacterial inclusion bodies with a recovery of 16.32 mg per liter of Luria broth medium. RESULTS: Before activation, the trypsinogen fusion protein did not show trypsin activity. Trypsinogen I was activated by adding 0.002 U of native TryI purified from the sardine pyloric caeca (nonrecombinant). The activated recombinant trypsin showed three times more activity than the nonrecombinant trypsin alone. CONCLUSION: The described protocol allowed obtaining sufficient amounts of recombinant TryI from Monterey sardine fish for further biochemical and biophysical characterization of its coldadaptation parameters.


Asunto(s)
Escherichia coli , Proteínas de Peces , Peces/genética , Cuerpos de Inclusión , Replegamiento Proteico , Tripsina , Animales , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Peces/biosíntesis , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/aislamiento & purificación , Cuerpos de Inclusión/química , Cuerpos de Inclusión/genética , Cuerpos de Inclusión/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Tripsina/biosíntesis , Tripsina/química , Tripsina/genética , Tripsina/aislamiento & purificación
16.
Food Res Int ; 111: 451-471, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-30007708

RESUMEN

Oxidative Stress (OS) produces the formation of free radicals and other reactive oxygen and nitrogen species that are intimately involved in many diseases, especially Chronic Degenerative Diseases (CDD) such as cancer, diabetes, cardiovascular diseases, and obesity, among others. Thus, reactive compounds need to be quenched by antioxidants. The problems of these compounds include that they are susceptible to degradation, have low bioavailability, and can lose their bioactivity in the gastroIntestinal tract. Therefore, an alternative is encapsulation. Zein is a protein used in nanotechnology as a polymer matrix because it can encapsulate different compounds such as antioxidants to provide stability and control of the release. The disadvantage of zein as a delivery vehicle is that it is limited by the low stability of aggregation when suspended in water, in addition to the conditions of acid pH or that higher ionic strength tends to destabilize. To reduce these limitations, the incorporation of polysaccharides as a second polymer matrix can provide stability in zein nanoparticles. In this review, we discuss OS as a source of CDD, the role of antioxidants in the prevention of these diseases, and the preparation, characterization, and application of antioxidant-zein-polysaccharide particles as delivery systems as well as possible mechanisms to control CDD.


Asunto(s)
Antioxidantes/farmacología , Nanopartículas/química , Polisacáridos/química , Zeína/química , Enfermedad Crónica , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Concentración de Iones de Hidrógeno , Concentración Osmolar , Estrés Oxidativo/efectos de los fármacos , Tamaño de la Partícula , Fenoles/farmacología , Polímeros/química
17.
Oxid Med Cell Longev ; 2017: 8219023, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29158873

RESUMEN

An imbalance between free radicals and antioxidants is known as oxidative stress, and it promotes cellular aging and the development of chronic noncommunicable diseases. The bioactive compounds present in food play an important role in preventing oxidative stress. The aim of this study was to determine the contributions and interactions of the hydroxycinnamic acids found in the bran and whole grain of sorghum and to evaluate their effects on the antioxidant capacity and inhibition of the hemolysis of human erythrocytes. Results showed that the caffeic acid, p-coumaric acid, and ferulic acid found in sorghum contributed to the scavenging of DPPH and ABTS radicals in various proportions. Ferulic acid, which was present in bound form in the bran and wholegrain sorghum, significantly inhibited the AAPH radical-induced oxidation of the erythrocyte membranes by 78.0 and 4.3%, respectively. Combinations of two, three, or four hydroxycinnamic acids may interact in an antagonistic or synergistic manner, thereby altering each other's bioactivities. The various interactions between the different sorghum bioactives can have a significant impact on their potential bioactivities. These results can be useful in the design of functional foods that aim to deliver bioactives to mitigate cellular aging or noncommunicable diseases.


Asunto(s)
Ácidos Cumáricos/metabolismo , Eritrocitos/efectos de los fármacos , Sorghum/metabolismo , Granos Enteros/metabolismo , Antioxidantes , Humanos , Oxidación-Reducción
18.
J Environ Sci Health B ; 51(11): 760-8, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27382921

RESUMEN

The aim of the present work was to evaluate the effect of mixtures of antifungal fractions extracted from Baccharis glutinosa and Jacquinia macrocarpa plants on the development of the filamentous fungi Aspergillus flavus and Fusarium verticillioides. The minimal inhibitory concentration that inhibited 50% of growth (MIC50) of each plant antifungal fraction was determined from the percentage radial growth inhibition of both fungi. Binomial mixtures made with both plant fractions were used at their MIC50 to determine the Fractional Inhibitory Concentration index (FIC index) for each fungus in order to evaluate their synergistic effect. Each synergistic mixture was analyzed in their effect on spore germination, spore size, spore viability, mitotic divisions, hyphal diameter and length, and number of septa per hypha. Some antifungal mixtures, even at low concentrations, showed higher antifungal effect than those of the individual antifungal fraction. The FIC indices of mixtures that showed the highest antifungal activity against A. flavus and F. verticillioides were 0.5272 and 0.4577, respectively, indicating a synergistic effect against both fungi. Only 12% and 8% of the spores of A. flavus and F. verticillioides, respectively, treated with the synergistic mixtures, were able to germinate, although their viability was not affected. An increase in the number of septa per hypha of both fungi was observed. The results indicated that the synergistic mixtures strongly affected the fungal growth even at lower concentrations than those of the individual plant fractions.


Asunto(s)
Antifúngicos/farmacología , Aspergillus flavus/efectos de los fármacos , Baccharis/química , Fusarium/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Primulaceae/química , México , Pruebas de Sensibilidad Microbiana
19.
Artículo en Inglés | MEDLINE | ID: mdl-27006678

RESUMEN

According to the World Health Organization, cancer is the main cause of mortality worldwide; thus, the search of chemopreventive compounds to prevent the disease has become a priority. White shrimp (Litopenaeus vannamei) has been reported as a source of compounds with chemopreventive activities. In this study, shrimp lipids were extracted and then fractionated in order to isolate those compounds responsible for the antimutagenic activity. The antimutagenic activity was assessed by the inhibition of the mutagenic effect of aflatoxin B1 on TA98 and TA100 Salmonella tester strains using the Ames test. Methanolic fraction was responsible for the highest antimutagenic activity (95.6 and 95.9% for TA98 and TA100, resp.) and was further separated into fifteen different subfractions (M1-M15). Fraction M8 exerted the highest inhibition of AFB1 mutation (96.5 and 101.6% for TA98 and TA100, resp.) and, after further fractionation, four subfractions M8a, M8b, M8c, and M8d were obtained. Data from (1)H and (13)C NMR, and mass spectrometry analysis of fraction M8a (the one with the highest antimutagenic activity), suggest that the compound responsible for its antimutagenicity is an apocarotenoid.

20.
Artículo en Inglés | MEDLINE | ID: mdl-26451153

RESUMEN

Vitex mollis is used in traditional Mexican medicine for the treatment of some ailments. However, there are no studies on what happens to the anti-inflammatory activity or antioxidant properties and total phenolic content of leaves and stem extracts of Vitex mollis during the digestion process; hence, this is the aim of this work. Methanolic, acetonic, and hexanic extracts were obtained from both parts of the plant. Extract yields and anti-inflammatory activity (elastase inhibition) were measured. Additionally, changes in antioxidant activity (DPPH and ABTS) and total phenols content of plant extracts before and after in vitro digestion were determined. The highest elastase inhibition to prevent inflammation was presented by hexanic extracts (leaf = 94.63% and stem = 98.30%). On the other hand, the major extract yield (16.14%), antioxidant properties (ABTS = 98.51% and DPPH = 94.47% of inhibition), and total phenols (33.70 mg GAE/g of dried sample) were showed by leaf methanolic extract. Finally, leaf and stem methanolic extracts presented an antioxidant activity increase of 35.25% and 27.22%, respectively, in comparison to their initial values after in vitro digestion process. All samples showed a decrease in total phenols at the end of the digestion. These results could be the basis to search for new therapeutic agents from Vitex mollis.

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