RESUMEN

The high content and quality of protein in Andean legumes make them valuable for producing protein hydrolysates using proteases from bacteria isolated from extreme environments. This study aimed to carry out a single-step purification of a haloprotease from Micrococcus sp. PC7 isolated from Peru salterns. In addition, characterize and apply the enzyme for the production of bioactive protein hydrolysates from underutilized Andean legumes. The PC7 protease was fully purified using only tangential flow filtration (TFF) and exhibited maximum activity at pH 7.5 and 40 °C. It was characterized as a serine protease with an estimated molecular weight of 130 kDa. PC7 activity was enhanced by Cu2+ (1.7-fold) and remained active in the presence of most surfactants and acetonitrile. Furthermore, it stayed completely active up to 6% NaCl and kept Ì´ 60% of its activity up to 8%. The protease maintained over 50% of its activity at 25 °C and 40 °C and over 70% at pH from 6 to 10 for up to 24 h. The determined Km and Vmax were 0.1098 mg mL-1 and 273.7 U mL-1, respectively. PC7 protease hydrolyzed 43%, 22% and 11% of the Lupinus mutabilis, Phaseolus lunatus and Erythrina edulis protein concentrates, respectively. Likewise, the hydrolysates from Lupinus mutabilis and Erythrina edulis presented the maximum antioxidant and antihypertensive activities, respectively. Our results demonstrated the feasibility of a simple purification step for the PC7 protease and its potential to be applied in industrial and biotechnological processes. Bioactive protein hydrolysates produced from Andean legumes may lead to the development of nutraceuticals and functional foods contributing to address some United Nations Sustainable Development Goals (SDGs).

Asunto(s)

Fabaceae , Micrococcus , Hidrolisados de Proteína , Micrococcus/metabolismo , Micrococcus/enzimología , Concentración de Iones de Hidrógeno , Hidrolisados de Proteína/química , Hidrolisados de Proteína/metabolismo , Peso Molecular , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/aislamiento & purificación , Perú , Temperatura , Serina Proteasas/metabolismo , Serina Proteasas/aislamiento & purificación , Serina Proteasas/química , Estabilidad de Enzimas , Cloruro de Sodio/metabolismo , Cloruro de Sodio/farmacología , Hidrólisis , Cinética

RESUMEN

Climate change intensifies soil salinization and jeopardizes the development of crops worldwide. The accumulation of salts in plant tissue activates the defense system and triggers ethylene production thus restricting cell division. We hypothesize that the inoculation of plant growth-promoting bacteria (PGPB) producing ACC (1-aminocyclopropane-1-carboxylate) deaminase favors the development of arbuscular mycorrhizal fungi (AMF), promoting the growth of maize plants under saline stress. We investigated the efficacy of individual inoculation of PGPB, which produce ACC deaminase, as well as the co-inoculation of PGPB with Rhizophagus clarus on maize plant growth subjected to saline stress. The isolates were acquired from the bulk and rhizospheric soil of Mimosa bimucronata (DC.) Kuntze in a temporary pond located in Pernambuco State, Brazil. In the first greenhouse experiment, 10 halophilic PGPB were inoculated into maize at 0, 40 and 80 mM of NaCl, and in the second experiment, the PGPB that showed the best performance were co-inoculated with R. clarus in maize under the same conditions as in the first experiment. Individual PGPB inoculation benefited the number of leaves, stem diameter, root and shoot dry mass, and the photosynthetic pigments. Inoculation with PGPB 28-10 Pseudarthrobacter enclensis, 24-1 P. enclensis and 52 P. chlorophenolicus increased the chlorophyll a content by 138%, 171%, and 324% at 0, 40 and 80 mM NaCl, respectively, comparing to the non-inoculated control. We also highlight that the inoculation of PGPB 28-10, 28-7 Arthrobacter sp. and 52 increased the content of chlorophyll b by 72%, 98%, and 280% and carotenoids by 82%, 98%, and 290% at 0, 40 and 80 mM of NaCl, respectively. Co-inoculation with PGPB 28-7, 46-1 Leclercia tamurae, 70 Artrobacter sp., and 79-1 Micrococcus endophyticus significantly increased the rate of mycorrhizal colonization by roughly 50%. Furthermore, co-inoculation promoted a decrease in the accumulation of Na and K extracted from plant tissue, with an increase in salt concentration, from 40 mM to 80 mM, also favoring the establishment and development of R. clarus. In addition, co-inoculation of these PGPB with R. clarus promoted maize growth and increased plant biomass through osmoregulation and protection of the photosynthetic apparatus. The tripartite symbiosis (plant-fungus-bacterium) is likely to reprogram metabolic pathways that improve maize growth and crop yield, suggesting that the AMF-PGPB consortium can minimize damages caused by saline stress.

Asunto(s)

Bacterias , Liasas de Carbono-Carbono , Micorrizas , Raíces de Plantas , Microbiología del Suelo , Zea mays , Zea mays/microbiología , Zea mays/crecimiento & desarrollo , Micorrizas/fisiología , Liasas de Carbono-Carbono/metabolismo , Raíces de Plantas/microbiología , Raíces de Plantas/crecimiento & desarrollo , Bacterias/clasificación , Bacterias/metabolismo , Bacterias/aislamiento & purificación , Estrés Salino , Clorofila/metabolismo , Glomeromycota/fisiología , Tolerancia a la Sal , Fotosíntesis , Rizosfera , Cloruro de Sodio/metabolismo , Hojas de la Planta/microbiología , Suelo/química

RESUMEN

Yeasts from cold environments have a wide range of strategies to prevent the negative effects of extreme conditions, including the production of metabolites of biotechnological interest. We investigated the growth profile and production of metabolites in yeast species isolated from cold environments. Thirty-eight strains were tested for their ability to grow at different temperatures (5-30 °C) and solute concentrations (3-12.5% NaCl and 50% glucose). All strains tested were able to grow at 5 °C, and 77% were able to grow with 5% NaCl at 18 °C. We were able to group strains based on different physicochemical/lifestyle profiles such as polyextremotolerant, osmotolerant, psychrotolerant, or psychrophilic. Five strains were selected to study biomass and metabolite production (glycerol, trehalose, ergosterol, and mycosporines). These analyses revealed that the accumulation pattern of trehalose and ergosterol was related to each lifestyle profile. Also, our findings would suggest that mycosporines does not have a role as an osmolyte. Non-conventional fermentative yeasts such as Phaffia tasmanica and Saccharomyces eubayanus may be of interest for trehalose production. This work contributes to the knowledge of non-conventional yeasts with biotechnological application from cold environments, including their growth profile, metabolites, and biomass production under different conditions.

Asunto(s)

Basidiomycota , Trehalosa , Trehalosa/metabolismo , Cloruro de Sodio/metabolismo , Levaduras , Ergosterol/metabolismo , Frío

RESUMEN

Castor (Ricinus communis) is a relevant industrial oilseed feedstock for many industrial applications, being globally mainly cultivated by smallholder farmers in semiarid areas, where abiotic stresses predominate. Therefore, susceptible to generating reactive oxygen species (ROS) and subsequent oxidative stress, compromising cell metabolism upon seed imbibition and germination, seedling and crop establishment, and yield. The present study evaluated the consequences of water restriction by Polyethylene glycol (PEG) and Sodium chloride (NaCl) on cell cycle and metabolism reactivation on germinability, seedling growth, and vigor parameters in 2 commercial castor genotypes (Nordestina and Paraguaçu). PEG water restriction inhibited germination completely at -0.23 MPa or higher, presumably due to reduced oxygen availability. The restrictive effects of NaCl saline stress on germination were observed only from -0.46 MPa onwards, affecting dry mass accumulation and the production of normal seedlings. In general, superoxide dismutase (SOD) activity increased in NaCl -0.23 MPa, whereas its modulation during the onset of imbibition (24h) seemed to depend on its initial levels in dry seeds in a genotype-specific manner, therefore, resulting in the higher stress tolerance of Nordestina compared to Paraguaçu. Overall, results show that Castor germination and seedling development are more sensitive to the restrictive effects of PEG than NaCl at similar osmotic potentials, contributing to a better understanding of the responses to water restriction stresses by different Castor genotypes. Ultimately, SOD may constitute a potential marker for characterizing castor genotypes in stressful situations during germination, early seedling, and crop establishment, and a target for breeding for Castor-improved stress tolerance.

Asunto(s)

Ricinus communis , Plantones , Plantones/metabolismo , Cloruro de Sodio/farmacología , Cloruro de Sodio/metabolismo , Ricinus communis/genética , Polietilenglicoles/farmacología , Polietilenglicoles/metabolismo , Germinación , Ciclo Celular , Semillas/metabolismo , Agua/metabolismo , Superóxido Dismutasa/metabolismo

RESUMEN

The renal Na-K-2Cl and Na-Cl cotransporters are the major salt reabsorption pathways in the thick ascending limb of Henle loop and the distal convoluted tubule, respectively. These transporters are the target of the loop and thiazide type diuretics extensively used in the world for the treatment of edematous states and arterial hypertension. The diuretics appeared in the market many years before the salt transport systems were discovered. The evolving of the knowledge and the cloning of the genes encoding the Na-K-2Cl and Na-Cl cotransporters were possible thanks to the study of marine species. This work presents the history of how we came to know the mechanisms for the loop and thiazide type diuretics actions, the use of marine species in the cloning process of these cotransporters and therefore in the whole solute carrier cotransproters 12 (SLC12) family of electroneutral cation chloride cotransporters, and the disease associated with each member of the family.

Asunto(s)

Cloruros , Simportadores de Cloruro de Sodio-Potasio , Animales , Humanos , Cationes/metabolismo , Cloruros/metabolismo , Diuréticos/metabolismo , Túbulos Renales Distales/metabolismo , Sodio/metabolismo , Cloruro de Sodio/metabolismo , Simportadores de Cloruro de Sodio-Potasio/genética , Simportadores de Cloruro de Sodio-Potasio/metabolismo , Miembro 3 de la Familia de Transportadores de Soluto 12/metabolismo , Tiazidas/metabolismo , Miembro 1 de la Familia de Transportadores de Soluto 12

RESUMEN

Extracellular proteases from halophilic archaea displays increased enzymatic activities in hypersaline environment. In this study, an extracellular protease-coding gene, hly34, from the haloarchaeal strain Halococcus salifodinae PRR34, was obtained through homologous search. The protease activity produced by this strain at 20% NaCl, 42 °C, and pH 7.0 was 32.5 ± 0.5 (U·mL-1). The codon-optimized hly34 which is specific for Escherichia coli can be expressed in E. coli instead of native hly34. It exhibits proteolytic activity under a wide range of low- or high-salt concentrations, slightly acidic or alkaline conditions, and slightly higher temperatures. The Hly34 presented the highest proteolytic activity at 50 °C, pH 9.0, and 0-1 M NaCl. It was found that the Hly34 showed a higher enzyme activity under low-salt conditions. Hly34 has good stability at different NaCl concentrations (1-4 M) and pH (6.0-10.0), as well as good tolerance to some metal ions. However, at 60 °C, the stability is reduced. It has a good tolerance to some metal ions. The proteolytic activity was completely inhibited by phenylmethanesulfonyl fluoride, suggesting that the Hly34 is a serine protease. This study further deepens our understanding of haloarchaeal extracellular protease, most of which found in halophilic archaea are classified as serine proteases. These proteases exhibit a certain level of alkaline resistance and moderate heat resistance, and they may emerge with higher activity under low-salt conditions than high-salt conditions. The protease Hly34 is capable of degrading a number of proteins, including substrate proteins, such as azocasein, whey protein and casein. It has promising applications in industrial production.

Asunto(s)

Halococcus , Halococcus/genética , Halococcus/metabolismo , Cloruro de Sodio/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Serina Proteasas , Serina Endopeptidasas , Metales , Iones , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Temperatura

RESUMEN

The primary structure of the thiazide-sensitive NaCl cotransporter (NCC) was resolved 30 years ago by the molecular identification of the cDNA encoding this cotransporter, from the winter's flounder urinary bladder, following a functional expression strategy. This review outlines some aspects of how the knowledge about thiazide diuretics and NCC evolved, the history of the cloning process, and the expansion of the SLC12 family of electroneutral cotransporters. The diseases associated with activation or inactivation of NCC are discussed, as well as the molecular model by which the activity of NCC is regulated. The controversies in the field are discussed as well as recent publication of the three-dimensional model of NCC obtained by cryo-electron microscopy, revealing not only the amino acid residues critical for Na+ and Cl- translocation but also the residues critical for polythiazide binding to the transporter, opening the possibility for a new era in thiazide diuretic therapy.

Asunto(s)

Proteínas Serina-Treonina Quinasas , Cloruro de Sodio , Miembro 3 de la Familia de Transportadores de Soluto 12/genética , Miembro 3 de la Familia de Transportadores de Soluto 12/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Cloruro de Sodio/metabolismo , Microscopía por Crioelectrón , Inhibidores de los Simportadores del Cloruro de Sodio , Clonación Molecular

RESUMEN

The Na+-activated Na+ channel (Nax) and salt-inducible kinase (SIK) are stimulated by increases in local Na+ concentration, affecting (Na+ + K+)-ATPase activity. To test the hypothesis that the triad Nax/SIK/(Na+ + K+)-ATPase contributes to kidney injury and salt-sensitive hypertension (HTN), uninephrectomized male Wistar rats (200 g; n = 20) were randomly divided into 4 groups based on a salt diet (normal salt diet; NSD-0.5% NaCl-or high-salt diet; HSD-4% NaCl) and subcutaneous administration of saline (0.9% NaCl) or deoxycorticosterone acetate (DOCA, 8 mg/kg), as follows: Control (CTRL), CTRL-Salt, DOCA, and DOCA-Salt, respectively. After 28 days, the following were measured: kidney function, blood pressure, (Na+ + K+)-ATPase and SIK1 kidney activities, and Nax and SIK1 renal expression levels. SIK isoforms in kidneys of CTRL rats were present in the glomerulus and tubular epithelia; they were not altered by HSD and/or HTN. CTRL-Salt rats remained normotensive but presented slight kidney function decay. HSD rats displayed augmentation of the Nax/SIK/(Na+ + K+)-ATPase pathway. HTN, kidney injury, and kidney function decay were present in all DOCA rats; these were aggravated by HSD. DOCA rats presented unaltered (Na+ + K+)-ATPase activity, diminished total SIK activity, and augmented SIK1 and Nax content in the kidney cortex. DOCA-Salt rats expressed SIK1 activity and downregulation in (Na+ + K+)-ATPase activity in the kidney cortex despite augmented Nax content. The data of this study indicate that the (Na+ + K+)-ATPase activity response to SIK is attenuated in rats under HSD, independent of HTN, as a mechanism contributing to kidney injury and salt-sensitive HTN.

Asunto(s)

Acetato de Desoxicorticosterona , Hipertensión , Ratas , Masculino , Animales , Cloruro de Sodio/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Ratas Wistar , Hipertensión/metabolismo , Sodio/metabolismo , Cloruro de Sodio Dietético/efectos adversos , Cloruro de Sodio Dietético/metabolismo , Presión Sanguínea , Riñón/metabolismo , Iones/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo

RESUMEN

Previously, we have shown that purinergic signalling is involved in the control of hyperosmotic-induced sympathoexcitation at the level of the PVN, via activation of P2X receptors. However, the source(s) of ATP that drives osmotically-induced increases in sympathetic outflow remained undetermined. Here, we tested the two competing hypotheses that either (1) higher extracellular ATP in PVN during salt loading (SL) is a result of a failure of ectonucleotidases to metabolize ATP; and/or (2) SL can stimulate PVN astrocytes to release ATP. Rats were salt loaded with a 2 % NaCl solution replacing drinking water up to 4 days, an experimental model known to cause a gradual increase in blood pressure and plasma osmolarity. Immunohistochemical assessment of glial-fibrillary acidic protein (GFAP) revealed increased glial cell reactivity in the PVN of rats after 4 days of high salt exposure. ATP and adenosine release measurements via biosensors in hypothalamic slices showed that baseline ATP release was increased 17-fold in the PVN while adenosine remained unchanged. Disruption of Ca2+-dependent vesicular release mechanisms in PVN astrocytes by virally-driven expression of a dominant-negative SNARE protein decreased the release of ATP. The activity of ectonucleotidases quantified in vitro by production of adenosine from ATP was increased in SL group. Our results showed that SL stimulates the release of ATP in the PVN, at least in part, from glial cells by a vesicle-mediated route and likely contributes to the neural control of circulation during osmotic challenges.

Asunto(s)

Núcleo Hipotalámico Paraventricular , Cloruro de Sodio , Ratas , Animales , Núcleo Hipotalámico Paraventricular/metabolismo , Cloruro de Sodio/metabolismo , Cloruro de Sodio Dietético/metabolismo , Astrocitos/metabolismo , Adenosina Trifosfato/metabolismo , Adenosina

RESUMEN

Biochemical and physiological parameters, growth, and yield of field crops especially salt sensitive crops like chickpea are affected adversely by salinity in arid to semi-arid regions. To investigate the effect of different salinity levels on growth, biochemical and physiological parameters of chickpea genotypes, a pot experiment following CRD, two factor factorial design, was conducted in the glasshouse at the Institute of Biotechnology and Genetic Engineering, The University of Agriculture, Peshawar, Pakistan. Ten (10) kg of soil was filled in each pot and salinity levels were maintained @ S0= 0 mM NaCl, S1= 50 mM NaCl, S2= 100 mM NaCl and S3= 150 mM by applying NaCl and 5 genotypes of chickpea (KK-2, Bhakkar-2011, Bittle-98, Punjab-2008, and CM-98) were used. At crop maturity, growth parameters, physiological, biochemical, and ionic parameters were measured using standard analysis procedures. Salinity reduced the growth and yield of all genotypes, but the rate of decrease was different among the genotypes tested. From the results, a decrease in K concentration, K/Na ratio, transpiration rate, stomatal conductance, N, and P was observed in all genotypes with the increase in salinity. An increase in salinity level increased the proline content (35.45%), crude protein (42%), H2O2 (19%), lipid peroxidation (62%), carbohydrates (23.22%), and Na+ concentration (137%). The highest level of salinity, 150 mM NaCl has exhibited the highest salinity stress in all parameters. Genotype KK-2 and Bhakkar-11 showed a lower rate of relative decrease in yield (4.5 and 12%), K+/Na+ ratio (23.34 and 11.47%), and K+ concentration (7.9 and 11%), respectively, and the lowest relative increase in Na+ accumulation (20.3 and 0.48%), @ 50 mM salinity compared to control. Genotype KK-2 and Bhakkar-11 proved better @ 50mM salinity. The findings suggest that the critical level of the salinity must be kept in mind and the salt-tolerant genotypes should be cultivated in salt affected soils.

Asunto(s)

Cicer , Salinidad , Cicer/genética , Cicer/metabolismo , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/farmacología , Cloruro de Sodio/metabolismo , Cloruro de Sodio/farmacología , Sodio/metabolismo , Sodio/farmacología , Genotipo , Suelo

RESUMEN

Cowpea (Vigna unguiculata (L.) Walp.) is a legume widely cultivated by small, medium and large producers in several Brazilian regions. However, one of the concerns for the production of cowpea in Brazil in recent years is the low rainfall activity in these regions, which generates the accumulation of salts on the surface. The objective of this work was to evaluate the effects of salt stress on growth parameters and enzyme activity in cowpea plants at different concentrations of brassinosteroids. Experiment was developed in a greenhouse using a completely randomized experimental design in a 3 x 3 factorial scheme. The treatments consisted of three levels of brassinosteroids (0, 3 and 6 µM EBL) and three levels of salt stress (0, 50 and 100 mM NaCl). Growth factors (height, diameter and number of leaves) decreased in the saline condition. With the presence of brassinosteroid the height did not increase, but the number of leaves did, mainly in the saline dosage of 100 mM NaCl. In the variable membrane integrity, brassinosteroid was efficient in both salinity dosages, the same not happening with the relative water content, where the saline condition did not affect the amount of water in the vegetable, with the application of brassino it remained high, decreasing only at dosage 100 mM NaCl. The nitrate reductase enzyme was greatly affected in the root system even with the application of increasing doses of brassino. Therefore, brassinosteroids as a promoter of saline tolerance in cowpea seedlings was positive. The concentration of 3µM of EBL provided the most satisfactory effect in tolerating the deleterious effects of the saline condition. The same cannot be concluded for the concentration of 6µM of EBL that did not promote tolerance to some variables.

Asunto(s)

Fabaceae , Vigna , Brasinoesteroides/metabolismo , Brasinoesteroides/farmacología , Estrés Salino , Cloruro de Sodio/metabolismo , Cloruro de Sodio/farmacología , Vigna/metabolismo , Agua/metabolismo

RESUMEN

Soil salinity is considered one of the main types of soil degradation in semiarid environments around the globe. This work aims to evaluate the effectiveness of soil conditioners to enhance the growth and salt extraction ability of Salicornia ramosíssima for different soil moisture contents. Salicornia plants were cultivated in pots in which the soils were treated with the following conditioners: control; gypsum + organic matter; elemental sulfur + organic matter; and gypsum + elemental sulfur + organic matter. Salicornia plants were subjected to two soil moisture rates - at 35 and 85% field capacity. Soil conditioners associated with higher contents of soil moisture promoted significant increases, compared to control, in fresh (6.20 - 11.13 g) and dry matter (1.20 - 2.07 g), relative biomass (100 - 179%) as well as significantly increased the concentrations of Na+ (56.09 - 65.64 mg kg-1) and Cl- (110.83 - 150.0 mg kg-1) in plant tissues. Soil conditioners significantly increased salt extraction ability under the two moisture levels, mainly by promoting higher values for both transfer factor and phytoremediation potential. The best performance of Salicornia in terms of plant yield and salt extraction, regardless of the moisture level, was the gypsum + organic matter.Novelty statementThere are no studies in the literature relating the use of conditioners as a strategy to enhance Salicornia's ability to extract salts.This work contributes to the management of salinized areas around the globe in two main aspects. The first is that many of these salt-degraded areas are desertified and through this study, it is possible to revegetate and recover them. The second one is that, since Salicornia is a plant with economic value, this can serve as an incentive for farmers to grow Salicornia in saline areas.

Asunto(s)

Chenopodiaceae , Suelo , Biodegradación Ambiental , Chenopodiaceae/metabolismo , Salinidad , Cloruro de Sodio/metabolismo

RESUMEN

Crude oil extracted from oilfield reservoirs brings together hypersaline produced water. Failure in pipelines transporting this mixture causes contamination of the soil with oil and hypersaline water. Soil salinization is harmful to biological populations, impairing the biodegradation of contaminants. We simulated the contamination of a soil from an oilfield with produced water containing different concentrations of NaCl and crude oil, in order to evaluate the effect of salinity and hydrocarbon concentration on prokaryote community structure and biodegradation activity. Microcosms were incubated in CO2-measuring respirometer. After the incubation, residual aliphatic hydrocarbons were quantified and were performed 16S rRNA gene sequencing. An increase in CO2 emission and hydrocarbon biodegradation was observed with increasing oil concentration up to 100 g kg-1. Alpha diversity decreased in oil-contaminated soils with an increase in the relative abundance of Actinobacteria and reduction of Bacteroidetes with increasing oil concentration. In the NaCl-contaminated soils, alpha diversity, CO2 emission, and hydrocarbon biodegradation decreased with increasing NaCl concentration. There was an increase in the relative abundance of Firmicutes and Proteobacteria and a reduction of Actinobacteria with increasing salt concentration. Our results highlight the need to adopt specific bioremediation strategies in soils impacted by mixtures of crude oil and hypersaline produced water.

Asunto(s)

Microbiota , Yacimiento de Petróleo y Gas/microbiología , Petróleo/metabolismo , Microbiología del Suelo , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Dióxido de Carbono/metabolismo , Hidrocarburos/metabolismo , Microbiota/genética , Petróleo/microbiología , ARN Ribosómico 16S/genética , Salinidad , Cloruro de Sodio/metabolismo , Suelo/química

RESUMEN

Saline environments are extreme habitats with a high diversity of microorganisms source of a myriad of biomolecules. These microorganisms are assigned as extremophiles recognized to be producers of new natural compounds, which can be synthesized by helping to survive under harshness and extreme conditions. In Brazil, in the saline and semi-arid region of Areia Branca (Caatinga biome), halotolerant bacteria (able to growth at high NaCl concentrations) were isolated from rhizosphere of native plants Blutaparon portulacoides and Spergularia sp. and their biopolymer production was studied. A total of 25 bacterial isolates were identified at genus level based on 16S rRNA gene sequence analysis. Isolates were mainly Gram-positive bacteria from Bacillaceae, Staphylococcaceae, Microbacteriaceae, and Bacillales XII incertae sedis families, affiliates to Bacillus, Staphylococcus, Curtobacterium, and Exiguobacterium genera, respectively. One of the Gram-negative isolates was identified as member of the Pseudomonadaceae family, genus Pseudomonas. All the identified strains were halotolerant bacteria with optimum growth at 0.6-2.0 M salt concentrations. Assays for biopolymer production showed that the halotolerant strains are a rich source of compounds as polyhydroxyalkanoates (PHA), biodegradable biopolymer, such as poly(3-hydroxybutyrate) (PHB) produced from low-cost substrates, and exopolysaccharides (EPS), such as hyaluronic acid (HA), metabolite of great interest to the cosmetic and pharmaceutical industry. Also, eight bacterial EPS extracts showed immunostimulatory activity, promising results that can be used in biomedical applications. Overall, our findings demonstrate that these biomolecules can be produced in culture medium with 0.6-2.0 M NaCl concentrations, relevant feature to avoid costly production processes. This is the first report of biopolymer-producing bacteria from a saline region of Caatinga biome that showed important biological activities.

Asunto(s)

Bacterias/aislamiento & purificación , Bacterias/metabolismo , Biopolímeros/metabolismo , Cloruro de Sodio/metabolismo , Microbiología del Suelo , Bacterias/clasificación , Bacterias/genética , Brasil , Filogenia , Polihidroxialcanoatos/metabolismo , Polisacáridos Bacterianos/metabolismo , Cloruro de Sodio/análisis , Suelo/química

RESUMEN

Description of a Gram-negative, motile, circular-shaped bacterial strain, designated A511T obtained from the skin of the pufferfish Sphoeroides spengleri (Family Tetraodontidae), collected in Arraial do Cabo, Brazil. Optimum growth occurs at 20-28 °C in the presence of 3% NaCl. The genome sequence of the novel isolate consisted of 4.36 Mb, 3,976 coding genes and G + C content of 42.5%. Genomic taxonomy analyses based on average amino acid (AAI), genome-to-genome-distance (GGDH) and phylogenetic reconstruction placed A511T (= CBAS 712T = CAIM 1939T) into a new species of the genus Vibrio (Vibrio tetraodonis sp. nov.). The genome of the novel species contains eight genes clusters (~ 183.9 Kbp in total) coding for different types of bioactive compounds that hint to several possible ecological roles in the pufferfish host.

Asunto(s)

Genoma Bacteriano/genética , Filogenia , Vibrio/clasificación , Vibrio/genética , Composición de Base , Brasil , ARN Ribosómico 16S/genética , Cloruro de Sodio/metabolismo , Especificidad de la Especie , Vibrio/crecimiento & desarrollo , Vibrio/metabolismo

RESUMEN

The use of plant growth-promoting bacteria represents an alternative to the massive use of mineral fertilizers in agriculture. However, some abiotic stresses commonly found in the environment, like salinity, can affect the efficiency of this approach. Here, we investigated the key mechanisms involved in the response of the plant growth-promoting bacterium Gluconacetobacter diazotrophicus to salt stress by using morphological and cell viability analyses, comparative proteomics, and reverse genetics. Our results revealed that the bacteria produce filamentous cells in response to salt at 100 mM and 150 mM NaCl. However, such a response was not observed at higher concentrations, where cell viability was severely affected. Proteomic analysis showed that salt stress modulates proteins involved in several pathways, including iron uptake, outer membrane efflux, osmotic adjustment, cell division and elongation, and protein transport and quality control. Proteomic data also revealed the repression of several extracytoplasmic proteins, especially those located at periplasm and outer membrane. The role of such pathways in the tolerance to salt stress was analyzed by the use of mutant defectives for Δtbdr (iron uptake), ΔmtlK and ΔotsA (compatible solutes synthesis), and ΔdegP (quality control of nascent extracytoplasmic proteins). ΔdegP presented the highest sensitivity to salt stress, Δtbdr, andΔmtlK also showed increased sensitivity, but ΔotsA was not affected. This is the first demonstration that DegP protein, a protease with minor chaperone activity, is essential for tolerance to salt stress in G. diazotrophicus. Our data contribute to a better understanding of the molecular bases that control the bacterial response/tolerance to salt stress, shedding light on quality control of nascent extracytoplasmic proteins.

Asunto(s)

Proteínas Bacterianas/metabolismo , Gluconacetobacter/metabolismo , Proteínas de Choque Térmico/metabolismo , Péptido Hidrolasas/metabolismo , Proteínas Periplasmáticas/metabolismo , Serina Endopeptidasas/metabolismo , Cloruro de Sodio/metabolismo , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Gluconacetobacter/enzimología , Gluconacetobacter/genética , Proteínas de Choque Térmico/genética , Hierro/metabolismo , Péptido Hidrolasas/genética , Proteínas Periplasmáticas/genética , Serina Endopeptidasas/genética

RESUMEN

We evaluated the temporal profile of the flavor enhancers monosodium glutamate (MSG), disodium inosinate (IMP), disodium guanylate (GMP), and monoammonium glutamate (MAG). We also evaluated the ability of these flavor enhancers to enhance salty taste in solutions containing different reductions of sodium chloride. Four experiments were conducted using Central Composite Rotational Design (CCRD) with focus on two objectives: concentration of flavor enhancers (0% to 1%) and reduction of sodium chloride content (0% to 100%). A 0.75% saline solution of NaCl was used as a control. In each experiment, the treatments were evaluated by the intensity of salty and umami tastes using an intensity scale. Treatments, selected according to the results of CCRD, were analyzed using time-intensity (TI) and temporal dominance of sensations (TDS) analyses. Glutamates (MSG/MAG) showed greater capacity to enhance salty taste than treatments containing nucleotides (IMP/GMP). The intensity of umami taste, using all the examined flavor enhancers, showed a similar sensory profile. Temporal perception curves (TI and TDS) of salty and umami tastes also showed a similar temporal profile. The glutamic acid amino acids were better able to improve salty taste than nucleotides in any range of sodium chloride reduction. Flavor enhancers showed greater ability to increase salty taste in smaller reductions in sodium chloride content. PRACTICAL APPLICATION: This research expand the knowledge about the ability to enhance the salty taste of flavor enhancers in different reductions in sodium content, Beside that, will provide information about the time profile of flavor enhancers. This study provides scientific technical information on the ability to intensify the salty taste of flavor enhancers and can assist the industry to develop new low sodium products and encourage the scientific community to conduct future research on this subject.

Asunto(s)

Aromatizantes/metabolismo , Inosina Monofosfato/metabolismo , Cloruro de Sodio/metabolismo , Glutamato de Sodio/metabolismo , Aromatizantes/análisis , Humanos , Inosina Monofosfato/análisis , Cloruro de Sodio/análisis , Glutamato de Sodio/análisis , Gusto

RESUMEN

Female sex hormones participate in the regulation of blood pressure and renal epithelial proliferation, effects not related to their reproductive function. About one-third of the world's population has abnormally high levels of blood pressure, hypertension, which is responsible for almost 50% of deaths from stroke and coronary heart disease. Salt sensitivity is a risk factor for cardiovascular morbidity and mortality and other diseases as well. We reported a model of salt sensitive hypertension in adult ovariectomized (oVx) Wistar rats. oVx rats are normotensive under normal salt intake (NS, 0.24% NaCl), but upon a high salt intake (HS, 1% NaCl) oVx rats developed a blood pressure profile of salt-sensitive hypertension. Our studies on kidney molecules related to sodium balance found that the circuit dopamine D1-like receptor, cytochrome P450 4A and Na+, K+-ATPase is altered by the absence of ovary hormones which is accompanied by a reduced ability to excrete sodium. In oVx rats HS intake also promotes changes in the expression of proteins related to sodium transport in peripheral blood mononuclear cells, mainly peripheral lymphocytes. Therefore, sodium transport is modified at several levels of normal physiology. Lately, we described that estradiol increases the rate of renal epithelial cell proliferation in primary cultures developed from human renal cortex. Thus, salt sensitivity, adaptive immunity, blood pressure and renal cell proliferation are complex biological responses regulated by female sex hormones.

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Un tercio de la población mundial tiene niveles anormalmente altos de presión arterial, hipertensión, responsable de casi el 50% de las muertes por accidente cerebrovascular y enfermedad coronaria. La sensibilidad a la sal es un factor de riesgo para la morbilidad y mortalidad cardiovascular y también para otras enfermedades. En estudios previos describimos un modelo de hipertensión sal sensible (HSS) en ratas Wistar ovariectomizadas (oVx) adultas. Las ratas oVx son normotensas con ingesta normal de sal (NS, 0.24% de NaCl), pero desarrollan un perfil de HSS con una ingesta elevada de sal (HS, 1% de NaCl). En los estudios en riñón encontramos que el circuito receptor D1 de dopamina, citocromo P450 4A y Na+, K+-ATPasa está alterado por la ausencia de hormonas ováricas, lo que se asocia a menor excreción de sodio e hipertensión arterial. La ingesta HS en ratas oVx también promueve cambios en la expresión de proteínas relacionadas con el transporte de sodio en células mononucleares de sangre periférica, principalmente linfocitos periféricos. Por lo tanto, el transporte de sodio se modifica en varios niveles de la fisiología normal. En estudios recientes observamos que el estradiol aumenta la proliferación y diferenciación de células epiteliales en cultivos de corteza renal humana. Sensibilidad a la sal, inmunidad adaptativa, presión arterial y proliferación de células epiteliales en riñón son fenómenos de gran importancia biológica regulados por estradiol.

Asunto(s)

Estradiol/metabolismo , Hipertensión/metabolismo , Riñón/metabolismo , Cloruro de Sodio/metabolismo , Animales , Presión Sanguínea , Proliferación Celular , Femenino , Humanos , Hipertensión/fisiopatología , Ratas , Ratas Wistar , Cloruro de Sodio/efectos adversos , ATPasa Intercambiadora de Sodio-Potasio

RESUMEN

Un tercio de la población mundial tiene niveles anormalmente altos de presión arterial, hipertensión, responsable de casi el 50% de las muertes por accidente cerebrovascular y enfermedad coronaria. La sensibilidad a la sal es un factor de riesgo para la morbilidad y mortalidad cardiovascular y también para otras enfermedades. En estudios previos describimos un modelo de hipertensión sal sensible (HSS) en ratas Wistar ovariectomizadas (oVx) adultas. Las ratas oVx son normotensas con ingesta normal de sal (NS, 0.24% de NaCl), pero desarrollan un perfil de HSS con una ingesta elevada de sal (HS, 1% de NaCl). En los estudios en riñón encontramos que el circuito receptor D1 de dopamina, citocromo P450 4A y Na+, K+-ATPasa está alterado por la ausencia de hormonas ováricas, lo que se asocia a menor excreción de sodio e hipertensión arterial. La ingesta HS en ratas oVx también promueve cambios en la expresión de proteínas relacionadas con el transporte de sodio en células mononucleares de sangre periférica, principalmente linfocitos periféricos. Por lo tanto, el transporte de sodio se modifica en varios niveles de la fisiología normal. En estudios recientes observamos que el estradiol aumenta la proliferación y diferenciación de células epiteliales en cultivos de corteza renal humana. Sensibilidad a la sal, inmunidad adaptativa, presión arterial y proliferación de células epiteliales en riñón son fenómenos de gran importancia biológica regulados por estradiol.

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Female sex hormones participate in the regulation of blood pressure and renal epithelial proliferation, effects not related to their reproductive function. About one-third of the world's population has abnormally high levels of blood pressure, hypertension, which is responsible for almost 50% of deaths from stroke and coronary heart disease. Salt sensitivity is a risk factor for cardiovascular morbidity and mortality and other diseases as well. We reported a model of salt sensitive hypertension in adult ovariectomized (oVx) Wistar rats. oVx rats are normotensive under normal salt intake (NS, 0.24% NaCl), but upon a high salt intake (HS, 1% NaCl) oVx rats developed a blood pressure profile of salt-sensitive hypertension. Our studies on kidney molecules related to sodium balance found that the circuit dopamine D1-like receptor, cytochrome P450 4A and Na+, K+-ATPase is altered by the absence of ovary hormones which is accompanied by a reduced ability to excrete sodium. In oVx rats HS intake also promotes changes in the expression of proteins related to sodium transport in peripheral blood mononuclear cells, mainly peripheral lymphocytes. Therefore, sodium transport is modified at several levels of normal physiology. Lately, we described that estradiol increases the rate of renal epithelial cell proliferation in primary cultures developed from human renal cortex. Thus, salt sensitivity, adaptive immunity, blood pressure and renal cell proliferation are complex biological responses regulated by female sex hormones.

Asunto(s)

Humanos , Animales , Femenino , Ratas , Cloruro de Sodio/metabolismo , Estradiol/metabolismo , Hipertensión/metabolismo , Riñón/metabolismo , Presión Sanguínea , Cloruro de Sodio/efectos adversos , Ratas Wistar , ATPasa Intercambiadora de Sodio-Potasio , Proliferación Celular , Hipertensión/fisiopatología

RESUMEN

New strategies to improve crop yield include the incorporation of plant growth-promoting bacteria in agricultural practices. The non-pathogenic bacterium Pseudomonas putida KT2440 is an excellent root colonizer of crops of agronomical importance and has been shown to activate the induced systemic resistance of plants in response to certain foliar pathogens. In this work, we have analyzed additional plant growth promotion features of this strain. We show it can tolerate high NaCl concentrations and determine how salinity influences traits such as the production of indole compounds, siderophore synthesis, and phosphate solubilization. Inoculation with P. putida KT2440 significantly improved seed germination and root and stem length of soybean and corn plants under saline conditions compared to uninoculated plants, whereas the effects were minor under non-saline conditions. Also, random transposon mutagenesis was used for preliminary identification of KT2440 genes involved in bacterial tolerance to saline stress. One of the obtained mutants was analyzed in detail. The disrupted gene encodes a predicted phosphoethanolamine-lipid A transferase (EptA), an enzyme described to be involved in the modification of lipid A during lipopolysaccharide (LPS) biosynthesis. This mutant showed changes in exopolysaccharide (EPS) production, low salinity tolerance, and reduced competitive fitness in the rhizosphere.

Asunto(s)

Proteínas Bacterianas/genética , Productos Agrícolas/microbiología , Desarrollo de la Planta , Raíces de Plantas/microbiología , Pseudomonas putida/fisiología , Estrés Salino , Productos Agrícolas/crecimiento & desarrollo , Etanolaminas/metabolismo , Pseudomonas putida/enzimología , Pseudomonas putida/genética , Rizosfera , Tolerancia a la Sal , Semillas/metabolismo , Cloruro de Sodio/metabolismo , Glycine max/metabolismo , Glycine max/microbiología , Transferasas/química , Transferasas/genética , Zea mays/metabolismo , Zea mays/microbiología