RESUMO
Gut microbiota members from the Bacteroidota phylum play a pivotal role in mammalian health and metabolism. They thrive in this diverse ecosystem due to their notable ability to cope with distinct recalcitrant dietary glycans via polysaccharide utilization loci (PULs). Our study reveals that a PUL from an herbivore gut bacterium belonging to the Bacteroidota phylum, with a gene composition similar to that in the human gut, exhibits extended functionality. While the human gut PUL targets mixed-linkage ß-glucans specifically, the herbivore gut PUL also efficiently processes linear and substituted ß-1,3-glucans. This gain of function emerges from molecular adaptations in recognition proteins and carbohydrate-active enzymes, including a ß-glucosidase specialized for ß(1,6)-glucosyl linkages, a typical substitution in ß(1,3)-glucans. These findings broaden the existing model for non-cellulosic ß-glucans utilization by gut bacteria, revealing an additional layer of functional and evolutionary complexity within the gut microbiota, beyond conventional gene insertions/deletions to intricate biochemical interactions.
Assuntos
Bacteroidetes , Microbioma Gastrointestinal , Herbivoria , beta-Glucanas , beta-Glucanas/metabolismo , Animais , Bacteroidetes/genética , Humanos , Filogenia , Metabolismo dos Carboidratos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismoRESUMO
There is a limited number of studies analyzing the molecular and biochemical processes regulating the metabolism of the maturation of Cocos nucifera L. zygotic embryos. Our research focused on the regulation of carbohydrate and lipid metabolic pathways occurring at three developmental stages of embryos from the Mexican Pacific tall (MPT) and the Yucatan green dwarf (YGD) cultivars. We used the TMT-synchronous precursor selection (SPS)-MS3 strategy to analyze the dynamics of proteomes from both embryos; 1044 and 540 proteins were determined for the MPT and YGD, respectively. A comparison of the differentially accumulated proteins (DAPs) revealed that the biological processes (BP) enriched in the MPT embryo included the glyoxylate and dicarboxylate metabolism along with fatty acid degradation, while in YGD, the nitrogen metabolism and pentose phosphate pathway were the most enriched BPs. Findings suggest that the MPT embryos use fatty acids to sustain a higher glycolytic/gluconeogenic metabolism than the YGD embryos. Moreover, the YGD proteome was enriched with proteins associated with biotic or abiotic stresses, e.g., peroxidase and catalase. The goal of this study was to highlight the differences in the regulation of carbohydrate and lipid metabolic pathways during the maturation of coconut YGD and MPT zygotic embryos.
Assuntos
Metabolismo dos Carboidratos , Cocos , Ácidos Graxos , Proteínas de Plantas , Sementes , Ácidos Graxos/metabolismo , Proteínas de Plantas/metabolismo , Sementes/metabolismo , Sementes/crescimento & desenvolvimento , Cocos/metabolismo , Proteômica/métodos , Proteoma/metabolismo , Metabolismo dos Lipídeos , Regulação da Expressão Gênica de PlantasRESUMO
Carbohydrates and fats constitute our primary energy sources. The importance of each of these energy substrates varies across cell types and physiological conditions. For example, the brain normally relies almost exclusively on glucose oxidation, whereas skeletal muscle shifts from lipids toward higher carbohydrate oxidation rates as exercise intensity increases. Understanding how carbohydrates are stored in our cells and which tissues contain significant carbohydrate stores is crucial for health professionals, especially given the role of carbohydrate metabolism in various pathophysiological conditions. This laboratory activity uses a simple and low-cost iodine binding method to quantify glycogen in mouse skeletal muscle and liver samples. By integrating the results of this activity with literature data, students can determine overall glycogen storage in the human body. The primary goal of the activity is to enhance students' understanding of the importance and limitations of glycogen stores in energy metabolism.NEW & NOTEWORTHY Carbohydrates are one of the primary energy sources utilized by our cells. Liver and skeletal muscle glycogen, which are the main carbohydrate reserves in the body, play a central role in energy metabolism, especially during periods of fasting and exercise. In this laboratory activity, students measure glycogen levels in tissues to gain insights into how carbohydrates are stored in our cells and understand the role and limitations of liver and muscle carbohydrate stores.
Assuntos
Glicogênio , Fígado , Músculo Esquelético , Fisiologia , Glicogênio/metabolismo , Animais , Fisiologia/educação , Músculo Esquelético/metabolismo , Humanos , Fígado/metabolismo , Camundongos , Metabolismo Energético/fisiologia , Metabolismo dos Carboidratos/fisiologia , LaboratóriosRESUMO
Nitrogen is the principal nutrient deficiency that increases lipids and carbohydrate content in diatoms but negatively affects biomass production. Marine diatom Chaetoceros muelleri is characterized by lipid and carbohydrate accumulation under low nitrogen concentration without affecting biomass. To elucidate the molecular effects of nitrogen concentrations, we performed an RNA-seq analysis of C. muelleri grown under four nitrogen concentrations (3.53 mM, 1.76 mM, 0.44 mM, and 0.18 mM of NaNO3). This research revealed that changes in global transcription in C. muelleri are differentially expressed by nitrogen concentration. "Energetic metabolism", "Carbohydrate metabolism" and "Lipid metabolism" pathways were identified as the most upregulated by N deficiency. Due to N limitation, alternative pathways to self-supply nitrogen employed by microalgal cells were identified. Additionally, nitrogen limitation decreased chlorophyll content and caused a greater response at the transcriptional level with a higher number of unigenes differentially expressed. By contrast, the highest N concentration (3.53 mM) recorded the lowest number of differentially expressed genes. Amt1, Nrt2, Fad2, Skn7, Wrky19, and Dgat2 genes were evaluated by RT-qPCR. In conclusion, C. muelleri modify their metabolic pathways to optimize nitrogen utilization and minimize nitrogen losses. On the other hand, the assembled transcriptome serves as the basis for metabolic engineering focused on improving the quantity and quality of the diatom for biotechnological applications. However, proteomic and metabolomic analysis is also required to compare gene expression, protein, and metabolite accumulation.
Assuntos
Diatomáceas , Nitrogênio , Transcriptoma , Nitrogênio/metabolismo , Diatomáceas/metabolismo , Diatomáceas/genética , Perfilação da Expressão Gênica/métodos , Metabolismo dos Lipídeos/genética , Metabolismo dos Carboidratos/genética , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/genética , BiomassaRESUMO
In this work extensive misconceptions of university students' -from nutrition area- about the metabolism of carbohydrates (CHM) in the human organism have been documented. The results lead to consider their difficulties concerning the learning of a complex set of imbricated biochemical models involved. Pursuant to these considerations, three physiological models are proposed as conceptual axes around which CHM in the human organism could be taught, in order to avoid fragmentation in students' knowledge and to give simple physiological contexts where to integrate those biochemical models. These contexts are: (a) a physiological model of the carbon cycle, (b) a physiological model of blood glucose uptake and homeostasis, and (c) a physiological model of the availability of small metabolites.
Assuntos
Metabolismo dos Carboidratos , Estudantes , Humanos , Aprendizagem , Carboidratos , EnsinoRESUMO
Aureobasidium pullulans LB83 is a versatile biocatalyst that produces a plethora of bioactive products thriving on a variety of feedstocks under the varying culture conditions. In our last study using this microorganism, we found cellulase activity (FPase, 2.27 U/ml; CMCase, 7.42 U/ml) and other plant cell wall degrading enzyme activities grown on sugarcane bagasse and soybean meal as carbon source and nitrogen, respectively. In the present study, we provide insights on the secretome analysis of this enzymatic cocktail. The secretome analysis of A. pullulans LB83 by Liquid Chromatography coupled to Mass Spectroscopy (LC-MS/MS) revealed 38 classes of Carbohydrate Active enZymes (CAZymes) of a total of 464 identified proteins. These CAZymes consisted of 21 glycoside hydrolases (55.26%), 12 glycoside hydrolases harboring carbohydrate-binding module (31.58%), 4 carbohydrate esterases (10.53%) and one glycosyl transferase (2.63%). To the best of our knowledge, this is the first report on the secretome analysis of A. pullulans LB83.
Assuntos
Aureobasidium , Metabolismo dos Carboidratos , Celulase , Celulose , Fermentação , Secretoma , Aureobasidium/enzimologia , Secretoma/enzimologia , Celulase/metabolismo , Saccharum/química , Cromatografia Líquida , Celulose/química , Celulose/metabolismoRESUMO
The use of a sprout suppressor is crucial for the use of potatoes beyond their natural dormancy period. The main sprout inhibitor used on a commercial scale, chlorpropham (CIPC), is becoming increasingly limited owing to its toxicity. Therefore, we evaluated the effectiveness of 1,4-dimethylnaphthalene (1,4-DMN) compared to CIPC in controlling sprouting and maintaining the quality of potato, Solanum tuberosum 'Asterix', during cold storage. Treatment with 1,4-DMN reduced fresh weight loss and controlled the number and length of sprouts comparable to CIPC. Compared to the control, both sprouting inhibitors led to higher starch and lower reducing sugar contents, and the tubers retained the recommended quality for industrial processing. After frying, less browning was observed in French fries obtained from 1,4-DMN- or CIPC-treated tubers. We ascertain that 1,4-DMN besides being an efficient sprouting inhibitor and alternative to CIPC, it contributes to maintaining the quality of French fries after cold storage.
Assuntos
Clorprofam , Solanum tuberosum , Clorprofam/metabolismo , Clorprofam/farmacologia , Solanum tuberosum/metabolismo , Naftalenos , Metabolismo dos Carboidratos , Tubérculos/metabolismoRESUMO
Temperature is an important factor that conditions the physiological responses of fish, generating a stressful condition when in non-ideal parameters. Thus, the objective was to evaluate metabolic aspects in the muscle of fish Astyanax lacustris submitted to thermal shock. The specimens were subjected to 2, 6, 12, 24, 48, and 96 h of exposure to 15 °C and 31 1 °C, with their respective controls 23 °C. At 15 °C there was a reduction in glycogenolysis in the initial periods indicated by changes in glycogen phosphorylase (GP), pyruvate and lactate. Subsequently, there was an increase in GP activity, pyruvate levels and hexokinase activity in the next time periods, suggesting an increase in energy demand. At 31 °C there was observed low need for the protein metabolism, indicated by reduction in the activity of aspartate aminotransferase and alanine aminotransferase. At 15 °C, initial periods show an increase in glutathione reductase activity and an increase in carbonylated protein levels, indicating induction of oxidative stress for muscle. At 31 °C, there was a punctual increase in reduced glutathione levels at 24 h. In addition, the integrated biomarker response index proved to be a good ally in the evaluation of a set of biomarkers, corroborating the results observed by the biomarkers individually. Thus, it is possible to conclude that the acute thermal shock affects the metabolism of A. lacustris muscle, which undergoes rearrangements to deal with temperature, where 15 °C is more stressful than 31 °C.
Assuntos
Antioxidantes , Músculos , Animais , Músculos/metabolismo , Antioxidantes/metabolismo , Estresse Oxidativo , Metabolismo dos Carboidratos , Biomarcadores/metabolismoRESUMO
La peroxidación lipídica es un proceso complejo que hace referencia a la degradación oxidativa de los lípidos, a través del cual los radicales libres capturan electrones de los lípidos en las membranas celulares, lo cual compromete la integridad y la función de la membrana. Mediante una serie de reacciones en cadena, se forman los peróxidos lipídicos que se degradan para formar compuestos reactivos como el malondialdehído (MDA) y 4-hidroxinonenal, los cuáles pueden ser cuantificados por diferentes metodologías. Objetivo: El presente trabajo se realizó con la finalidad establecer el grado de oxidación en una población con diabetes tipo 2 (DM2).Métodos: Estudio descriptivo, analítico y transversal; muestra de 55 personas, conformada por 30 controles entre 25-35 años y 25 pacientes con DM2 entre 25-50 años, se les determinó glicemia, triglicéridos, colesterol total, HDL-Colesterol y LDL-Colesterol por método colorimétrico enzimático, así como se determinó la concentración de 4-hidroxinonenal como un marcador de estrés oxidativo Resultados: Los valores de 4-hidroxinonenal en la población control oscilaron entre 2,61y 6,83 µmol/L y en los diabéticos de 28,99 y 73,74 µmol/L., encontrándose diferencias estadísticamente significativas entre ambas poblaciones, así como en el perfil lipídico y en la glicemia entre ambos grupos. Conclusión: Los resultados demuestran una elevación de la peroxidación lipídica en pacientes diabéticos, lo cual es indicativo de estrés oxidativo y riesgo adicional en estos pacientes que podrían conllevar a las complicaciones crónicas dela diabetes tipo 2(AU)
Lipid peroxidation is a complexprocess that refers to the oxidative degradation of lipids, through which free radicals capture electrons from lipids incell membranes, which compromises the integrity and functionof the membrane. Trough a series of chain reactions, lipidperoxides are formed that degrade to form reactive compoundssuch as malondialdehyde (MDA) and 4-hydroxynonenal, whichcan be quantified by different methodologies. Objective: The present work was carried out with the purpose ofestablishing the degree of oxidation in a population withtype 2 diabetes (DM2). Methods: the sample was 55 people,made up of 30 controls between 25-35 years and 25 patientswith DM2 and between 25-50 years, glycemia, triglycerides,total cholesterol, HDL-Cholesterol and LDL-Cholesterol were etermined by colorimetric method. enzymatic, as well as theconcentration of 4-hydroxynonenal was determined as a markerof oxidative stress. Results: The values of 4-hydroxynonenal inthe control population ranged between 2.61 and 6.83 µmol/Land in diabetics 28.99 and 73.74 µmol/L., finding statisticallysignificant differences between both populations, as well as inthe lipid profile and glycemia between both groups. Conclusion:The results show an elevation of lipid peroxidation in diabeticpatients, which is indicative of oxidative stress and additionalrisk in these patients that could lead to chronic complications oftype 2 diabetes(AU)
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Peroxidação de Lipídeos , Diabetes Mellitus Tipo 2 , Oxidação , Triglicerídeos , Glicemia , Colesterol , Metabolismo dos CarboidratosRESUMO
The largest living rodent, capybara, can efficiently depolymerize and utilize lignocellulosic biomass through microbial symbiotic mechanisms yet elusive. Herein, we elucidate the microbial community composition, enzymatic systems and metabolic pathways involved in the conversion of dietary fibers into short-chain fatty acids, a main energy source for the host. In this microbiota, the unconventional enzymatic machinery from Fibrobacteres seems to drive cellulose degradation, whereas a diverse set of carbohydrate-active enzymes from Bacteroidetes, organized in polysaccharide utilization loci, are accounted to tackle complex hemicelluloses typically found in gramineous and aquatic plants. Exploring the genetic potential of this community, we discover a glycoside hydrolase family of ß-galactosidases (named as GH173), and a carbohydrate-binding module family (named as CBM89) involved in xylan binding that establishes an unprecedented three-dimensional fold among associated modules to carbohydrate-active enzymes. Together, these results demonstrate how the capybara gut microbiota orchestrates the depolymerization and utilization of plant fibers, representing an untapped reservoir of enzymatic mechanisms to overcome the lignocellulose recalcitrance, a central challenge toward a sustainable and bio-based economy.
Assuntos
Microbioma Gastrointestinal , Plantas/metabolismo , Polissacarídeos/metabolismo , Roedores/microbiologia , Animais , Bactérias/classificação , Bactérias/enzimologia , Bactérias/metabolismo , Bacteroidetes/enzimologia , Bacteroidetes/genética , Bacteroidetes/metabolismo , Metabolismo dos Carboidratos , Cristalografia por Raios X , Fibras na Dieta/metabolismo , Glicosídeo Hidrolases/metabolismo , Lignina , Filogenia , Simbiose , Xilanos/metabolismoRESUMO
Penicillium echinulatum 2HH is an ascomycete well known for its production of cellulolytic enzymes. Understanding lignocellulolytic and sugar uptake systems is essential to obtain efficient fungi strains for the production of bioethanol. In this study we performed a genome-wide functional annotation of carbohydrate-active enzymes and sugar transporters involved in the lignocellulolytic system of P. echinulatum 2HH and S1M29 strains (wildtype and mutant, respectively) and eleven related fungi. Additionally, signal peptide and orthology prediction were carried out. We encountered a diverse assortment of cellulolytic enzymes in P. echinulatum, especially in terms of ß-glucosidases and endoglucanases. Other enzymes required for the breakdown of cellulosic biomass were also found, including cellobiohydrolases, lytic cellulose monooxygenases and cellobiose dehydrogenases. The S1M29 mutant, which is known to produce an increased cellulase activity, and the 2HH wild type strain of P. echinulatum did not show significant differences between their enzymatic repertoire. Nevertheless, we unveiled an amino acid substitution for a predicted intracellular ß-glucosidase of the mutant, which might contribute to hyperexpression of cellulases through a cellodextrin induction pathway. Most of the P. echinulatum enzymes presented orthologs in P. oxalicum 114-2, supporting the presence of highly similar cellulolytic mechanisms and a close phylogenetic relationship between these fungi. A phylogenetic analysis of intracellular ß-glucosidases and sugar transporters allowed us to identify several proteins potentially involved in the accumulation of intracellular cellodextrins. These may prove valuable targets in the genetic engineering of P. echinulatum focused on industrial cellulases production. Our study marks an important step in characterizing and understanding the molecular mechanisms employed by P. echinulatum in the enzymatic hydrolysis of lignocellulosic biomass.
Assuntos
Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Lignina/metabolismo , Penicillium/metabolismo , Substituição de Aminoácidos , Transporte Biológico , Metabolismo dos Carboidratos , Celulose/análogos & derivados , Dextrinas , Regulação Fúngica da Expressão Gênica , Anotação de Sequência Molecular , Penicillium/genética , Filogenia , Açúcares/metabolismoRESUMO
Carbohydrate metabolism via cyclodextrins (CM-CD) is an uncommon starch-converting pathway that thoroughly depends on extracellular cyclomaltodextrin glucanotransferases (CGTases) to transform the surrounding starch substrate to α-(1,4)-linked oligosaccharides and cyclodextrins (CDs). The CM-CD pathway has emerged as a convenient microbial adaptation to thrive under extreme temperatures, as CDs are functional amphipathic toroids with higher heat-resistant values than linear dextrins. Nevertheless, although the CM-CD pathway has been described in a few mesophilic bacteria and archaea, it remains obscure in extremely thermophilic prokaryotes (Topt ≥ 70 °C). Here, a new monophyletic group of CGTases with an exceptional three-domain ABC architecture was detected by (meta)genome mining of extremely thermophilic Thermoanaerobacterales living in a wide variety of hot starch-poor environments on Earth. Functional studies of a representative member, CldA, showed a maximum activity in a thermoacidophilic range (pH 4.0 and 80 °C) with remarkable product diversification that yielded a mixture of α:ß:γ-CDs (34:62:4) from soluble starch, as well as G3-G7 linear dextrins and fermentable sugars as the primary products. Together, comparative genomics and predictive functional analysis, combined with data of the functionally characterized key proteins of the gene clusters encoding CGTases, revealed the CM-CD pathway in Thermoanaerobacterales and showed that it is involved in the synthesis, transportation, degradation, and metabolic assimilation of CDs.
Assuntos
Metabolismo dos Carboidratos/fisiologia , Ciclodextrinas/metabolismo , Glucosiltransferases/genética , Glucosiltransferases/fisiologia , Thermoanaerobacterium/metabolismo , Genoma Bacteriano/genética , Glucosiltransferases/metabolismo , Família Multigênica , Thermoanaerobacterium/genéticaRESUMO
Flooding negatively influences the growth and development of several plant species. Here, we show that the flood tolerance of young Handroanthus chrysotrichus plants involves growth deficit, carbon assimilation reductions, starch remobilization, and energy regulation. The effect of hypoxia was evaluated in a controlled experiment consisting of plants subjected to normoxia and water-logging, with later recovery. We measured morphological changes, gas exchange, photosynthetic pigments, soluble carbohydrates and starch contents, the activity of the enzymes alcohol dehydrogenase (ADH), and pyruvate decarboxylase (PDC), and ATP and ADP levels. While control plants showed normal appearance and growth, flooded plants exhibited a drastic decrease in growth, necrosis of some root tips, hypertrophic lenticels on the stems, and foliar chlorosis. Oxygen deprivation in root cells led to a significant decrease in stomatal conductance. The low Amax rates caused a decline in foliar soluble sugar content at 20 days and a subsequent increase in the leaves and roots, coinciding with starch degradation at 40 days. We also observed increases of 220.5% in ADH and 292% in PDC activities in the roots at 20 and 40 days of flooding. The activation of anaerobic metabolism in stressed plants was an essential mechanism for ATP regulation in both tissues used to maintain a minimal metabolism to cope with hypoxia to the detriment of growth. The post-stress recovery process in H. chrysotrichus occurred slowly, with gas exchange gradually resumed and anaerobic metabolism and sugar content maintained to improve energy regulation.
Assuntos
Inundações , Raízes de Plantas , Metabolismo dos Carboidratos , Fotossíntese/fisiologia , Folhas de Planta/fisiologia , Raízes de Plantas/metabolismoRESUMO
MAIN CONCLUSION: The relationship of fructan to plant growth regulators is clearly more complicated than it looks and is likely related to differences between fructan molecules in size and structure as well as localization. Fructans are a complex group of carbohydrates composed mainly of fructose units linked to a sucrose molecule. Fructans are present in plants as heterogeneous mixtures with diverse molecular structures and mass, different polymerization degrees, and linkage types between fructosyl residues. Like sucrose, they are frequently stored in leaves and other organs, acting as carbohydrate reserves. Fructans are synthesized in the cell vacuole by fructosyltransferase enzymes and catabolized by fructan exohydrolase enzymes. Several publications have shown that fructan metabolism varies with the stage of plant development and in response to the environment. Recent studies have shown a correlation between plant growth regulators (PGR), fructan metabolism, and tolerance to drought and cold. PGR are compounds that profoundly influence the growth and differentiation of plant cells, tissues, and organs. They play a fundamental role in regulating plant responses to developmental and environmental signals. In this review, we summarize the most up-to-date knowledge on the metabolism of fructans and their crosstalk with PGR signaling pathways. We identify areas that require more research to complete our understanding of the role of fructans in plants.
Assuntos
Hexosiltransferases , Reguladores de Crescimento de Plantas , Metabolismo dos Carboidratos , Frutanos , Hexosiltransferases/metabolismo , Folhas de Planta/metabolismo , SacaroseRESUMO
KEY MESSAGE: The functional absence of the electron-transfer flavoprotein: ubiquinone oxidoreductase (ETFQO) directly impacts electrons donation to the mitochondrial electron transport chain under carbohydrate-limiting conditions without major impacts on the respiration of cell cultures. Alternative substrates (e.g., amino acids) can directly feed electrons into the mitochondrial electron transport chain (mETC) via the electron transfer flavoprotein/electron-transfer flavoprotein: ubiquinone oxidoreductase (ETF/ETFQO) complex, which supports plant respiration during stress situations. By using a cell culture system, here we investigated the responses of Arabidopsis thaliana mutants deficient in the expression of ETFQO (etfqo-1) following carbon limitation and supplied with amino acids. Our results demonstrate that isovaleryl-CoA dehydrogenase (IVDH) activity was induced during carbon limitation only in wild-type and that these changes occurred concomit with enhanced protein content. By contrast, neither the activity nor the total amount of IVDH was altered in etfqo-1 mutants. We also demonstrate that the activities of mitochondrial complexes in etfqo-1 mutants, display a similar pattern as in wild-type cells. Our findings suggest that the defect of ETFQO protein culminates with an impaired functioning of the IVDH, since no induction of IVDH activity was observed. However, the functional absence of the ETFQO seems not to cause major impacts on plant respiration under carbon limiting conditions, most likely due to other alternative electron entry pathways.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Flavoproteínas Transferidoras de Elétrons , Aminoácidos de Cadeia Ramificada/farmacologia , Arabidopsis/citologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Metabolismo dos Carboidratos , Técnicas de Cultura de Células , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Flavoproteínas Transferidoras de Elétrons/genética , Flavoproteínas Transferidoras de Elétrons/metabolismo , Regulação da Expressão Gênica de Plantas , Isovaleril-CoA Desidrogenase/genética , Isovaleril-CoA Desidrogenase/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , MutaçãoRESUMO
Humicola grisea var. thermoidea is a thermophilic ascomycete and important enzyme producer that has an efficient enzymatic system with a broad spectrum of thermostable carbohydrate-active (CAZy) enzymes. These enzymes can be employed in lignocellulose biomass deconstruction and other industrial applications. In this work, the genome of H. grisea var. thermoidea was sequenced. The acquired sequence reads were assembled into a total length of 28.75 Mbp. Genome features correlate with what was expected for thermophilic Sordariomycetes. The transcriptomic data showed that sugarcane bagasse significantly upregulated genes related to primary metabolism and polysaccharide deconstruction, especially hydrolases, at both pH 5 and pH 8. However, a number of exclusive and shared genes between the pH values were found, especially at pH 8. H. grisea expresses an average of 211 CAZy enzymes (CAZymes), which are capable of acting in different substrates. The top upregulated genes at both pH values represent CAZyme-encoding genes from different classes, including acetylxylan esterase, endo-1,4-ß-mannosidase, exoglucanase, and endoglucanase genes. For the first time, the arsenal that the thermophilic fungus H. grisea var. thermoidea possesses to degrade the lignocellulosic biomass is shown. Carbon source and pH are of pivotal importance in regulating gene expression in this organism, and alkaline pH is a key regulatory factor for sugarcane bagasse hydrolysis. This work paves the way for the genetic manipulation and robust biotechnological applications of this fungus. IMPORTANCE Most studies regarding the use of fungi as enzyme producers for biomass deconstruction have focused on mesophile species, whereas the potential of thermophiles has been evaluated less. This study revealed, through genome and transcriptome analyses, the genetic repertoire of the biotechnological relevant thermophile fungus Humicola grisea. Comparative genomics helped us to further understand the biology and biotechnological potential of H. grisea. The results demonstrate that this fungus possesses an arsenal of carbohydrate-active (CAZy) enzymes to degrade the lignocellulosic biomass. Indeed, it expresses more than 200 genes encoding CAZy enzymes when cultivated in sugarcane bagasse. Carbon source and pH are key factors for regulating the gene expression in this organism. This work shows, for the first time, the great potential of H. grisea as an enzyme producer and a gene donor for biotechnological applications and provides the base for the genetic manipulation and robust biotechnological applications of this fungus.
Assuntos
Ascomicetos/enzimologia , Ascomicetos/metabolismo , Metabolismo dos Carboidratos/fisiologia , Lignina/metabolismo , Saccharum/microbiologia , Ascomicetos/genética , Composição de Bases/genética , Biomassa , Metabolismo dos Carboidratos/genética , Perfilação da Expressão Gênica , Genoma Fúngico/genética , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Saccharum/metabolismo , Transcriptoma/genética , Sequenciamento Completo do GenomaRESUMO
Although lignocellulose is the most abundant and renewable natural resource for biofuel production, its use remains under exploration because of its highly recalcitrant structure. Its deconstruction into sugar monomers is mainly driven by carbohydrate-active enzymes (CAZymes). To develop highly efficient and fast strategies to discover biomass-degrading enzymes for biorefinery applications, an enrichment process combined with integrative omics approaches was used to identify new CAZymes. The lignocellulolytic-enriched mangrove microbial community (LignoManG) established on sugarcane bagasse (SB) was enriched with lignocellulolytic bacteria and fungi such as Proteobacteria, Bacteroidetes, Basidiomycota, and Ascomycota. These microbial communities were able to degrade up to 55 % of the total SB, indicating the production of lignocellulolytic enzymes. Metagenomic analysis revealed that the LignoManG harbors 18.042 CAZyme sequences such as of cellulases, hemicellulases, carbohydrate esterases, and lytic polysaccharide monooxygenase. Similarly, our metaproteomic analysis depicted several enzymes from distinct families of different CAZy families. Based on the LignoManG data, a xylanase (coldXynZ) was selected, amplified, cloned, expressed, and biochemically characterized. The enzyme displayed psicrofilic properties, with the highest activity at 15⯰C, retaining 77 % of its activity when incubated at 0⯰C. Moreover, molecular modeling in silico indicated that coldXynZ is composed of a TIM barrel, which is a typical folding found in the GH10 family, and displayed similar structural features related to cold-adapted enzymes. Collectively, the data generated in this study represent a valuable resource for lignocellulolytic enzymes with potential biotechnological applications.
Assuntos
Ascomicetos , Celulases , Saccharum , Bacteroidetes , Basidiomycota , Biomassa , Metabolismo dos Carboidratos , Esterases , Sedimentos Geológicos , Glicosídeo Hidrolases , Metagenoma , Proteobactérias , Áreas AlagadasRESUMO
The Bifidobacterium longum 51A strain of isolated from feces of a healthy child, has demonstrated probiotic properties by in vivo and in vitro studies, which may be assigned to its production of metabolites such as acetate. Thus, through the study of comparative genomics, the present work sought to identify unique genes that might be related to the production of acetate. To perform the study, the DNA strain was sequenced using Illumina HiSeq technology, followed by assembly and manual curation of coding sequences. Comparative analysis was performed including 19 complete B. longum genomes available in Genbank/NCBI. In the phylogenetic analysis, the CECT 7210 and 157F strains of B. longum subsp. infantis aggregated within the subsp. longum cluster, suggesting that their taxonomic classification should be reviewed. The strain 51A of B. longum has 26 unique genes, six of which are possibly related to carbohydrate metabolism and acetate production. The phosphoketolase pathway from B. longum 51A showed a difference in acetyl-phosphate production. This result seems to corroborate the analysis of their unique genes, whose presence suggests the strain may use different sources of carbohydrates that allow a greater production of acetate and consequently offer benefits to the host health.
Assuntos
Acetatos/metabolismo , Bifidobacterium longum/genética , Bifidobacterium longum/metabolismo , Metabolismo dos Carboidratos/genética , Genes Bacterianos , Probióticos/metabolismo , Sequência de Bases , Bifidobacterium longum/classificação , Criança , Simulação por Computador , Fezes/microbiologia , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Filogenia , Análise de Sequência de DNARESUMO
Introducción: La asociación de asma y obesidad ha aumentado su prevalencia en los últimos años. Objetivos: Caracterizar a niños y adolescentes asmáticos con obesidad asociada. Métodos: Investigación descriptiva, transversal y prospectiva, 2017-2018, Servicio de Neumología, Hospital Pediátrico Universitario Centro Habana. Se evaluaron 43 pacientes asmáticos entre 5 y 18 años de edad con obesidad exógena asociada. Se exploraron edad, sexo, antecedentes familiares de asma y obesidad, severidad y grado de control del asma. Se realizaron estudios para evaluar el metabolismo lipídico y de los hidratos de carbono. Resultados: La distribución por edades fue similar (escolares, adolescencia temprana y tardía), predominó el sexo masculino en escolares: 11 de 16 (68,7 por ciento) y el femenino en la adolescencia tardía: 12 de 15 (80 por ciento). Se refirieron antecedentes familiares de obesidad en 5 (11,6 por ciento) y asma asociada a obesidad en 26 (60,5 por ciento); la mayoría presentaba asma persistente leve: 38 (88,4 por ciento), no controlados: 28 (65,1 por ciento) y parcialmente controlados 10 (23,3 por ciento). Dos pacientes presentaban intolerancia a los hidratos de carbono (4,7 por ciento), uno de ellos con resistencia a la insulina. La dislipidemia fue el trastorno metabólico más frecuente: colesterol sérico elevado en 23 (53,5 por ciento), LDLc alto en 13 (30,2 por ciento) y HDLc bajo en 10 (23,3 por ciento). Conclusiones: Los niños asmáticos con historia familiar de obesidad tienen alto riesgo de ser también obesos, lo cual puede dificultar el control del asma; son frecuentes en ellos las alteraciones metabólicas(AU)
Introduction: The association of asthma and obesity has increased its prevalence in recent years. Objectives: Characterize asthmatic children and adolescents with associated obesity. Methods: Descriptive, cross-sectional and prospective research, 2017 - 2018, Pneumology Service, Centro Habana University Pediatric Hospital. 43 asthmatic patients from 5 to 18 years old with associated exogenous obesity were assessed. Age, sex, family history of asthma and obesity, severity and degree of asthma control were studied. Studies were conducted to evaluate lipid and carbohydrate metabolism. Results: Age distribution was similar (schoolers, early and late adolescence), predominated male sex in schoolchildren (11 of 16 - 68.7 percent) and the female in late adolescence (12 of 15 -80 percent). Family history of obesity was reported in 5 patients (11.6 percent) and obesity-associated asthma in 26 (60.5 percent); most had mild persistent asthma (38 -88.4 percent) poorly controlled: 28 uncontrolled (65.1 percent) and 10 partially controlled (23.3 percent). Two patients had carbohydrate intolerance (4.7 percent), one of them with insulin resistance. Dyslipidemia was the most common metabolic disorder: serum cholesterol increased in 23 - 53.5 percent, high LDLc in 13 - 30.2 percent, and low HDLc in 10 - 23.3 percent. Conclusions: Asthmatic children with a family history of obesity are also at high risk of being obese, which can make difficult to control asthma; metabolic alterations are common in them(Au)
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Adolescente , Asma , Distribuição por Idade , Dislipidemias , Metabolismo dos Carboidratos , ObesidadeRESUMO
This review commemorates the 50th anniversary of the Nobel Prize in Chemistry awarded to Luis F. Leloir 'for his discovery of sugar-nucleotides and their role in the biosynthesis of carbohydrates'. He and his co-workers discovered that activated forms of simple sugars, such as UDP-glucose and UDP-galactose, are essential intermediates in the interconversion of sugars. They elucidated the biosynthetic pathways for sucrose and starch, which are the major end-products of photosynthesis, and for trehalose. Trehalose 6-phosphate, the intermediate of trehalose biosynthesis that they discovered, is now a molecule of great interest due to its function as a sugar signalling metabolite that regulates many aspects of plant metabolism and development. The work of the Leloir group also opened the doors to an understanding of the biosynthesis of cellulose and other structural cell wall polysaccharides (hemicelluloses and pectins), and ascorbic acid (vitamin C). Nucleotide-sugars also serve as sugar donors for a myriad of glycosyltransferases that conjugate sugars to other molecules, including lipids, phytohormones, secondary metabolites, and proteins, thereby modifying their biological activity. In this review, we highlight the diversity of nucleotide-sugars and their functions in plants, in recognition of Leloir's rich and enduring legacy to plant science.