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1.
Nat Commun ; 11(1): 5104, 2020 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-33037214

RESUMO

Many intestinal pathogens, including Clostridioides difficile, use mucus-derived sugars as crucial nutrients in the gut. Commensals that compete with pathogens for such nutrients are therefore ecological gatekeepers in healthy guts, and are attractive candidates for therapeutic interventions. Nevertheless, there is a poor understanding of which commensals use mucin-derived sugars in situ as well as their potential to impede pathogen colonization. Here, we identify mouse gut commensals that utilize mucus-derived monosaccharides within complex communities using single-cell stable isotope probing, Raman-activated cell sorting and mini-metagenomics. Sequencing of cell-sorted fractions reveals members of the underexplored family Muribaculaceae as major mucin monosaccharide foragers, followed by members of Lachnospiraceae, Rikenellaceae, and Bacteroidaceae families. Using this information, we assembled a five-member consortium of sialic acid and N-acetylglucosamine utilizers that impedes C. difficile's access to these mucosal sugars and impairs pathogen colonization in antibiotic-treated mice. Our findings underscore the value of targeted approaches to identify organisms utilizing key nutrients and to rationally design effective probiotic mixtures.


Assuntos
/patogenicidade , Microbioma Gastrointestinal/fisiologia , Monossacarídeos/metabolismo , Acetilglucosamina/metabolismo , Animais , Antibacterianos , Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Separação Celular/métodos , /crescimento & desenvolvimento , Infecções por Clostridium/microbiologia , Deutério , Feminino , Mucinas Gástricas/química , Mucinas Gástricas/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/microbiologia , Metagenoma , Camundongos Endogâmicos C57BL , Ácido N-Acetilneuramínico/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismo , Análise Espectral Raman
2.
Gene ; 741: 144522, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32145329

RESUMO

Virus-induced gene silencing (VIGS) is a transient based reverse genetic tool used to elucidate the function of novel gene in N. benthamiana. In current study, 14 UDP-D-glucuronate 4-epimerase (GAE) family members were identified and their gene structure, phylogeny and expression pattern were analyzed. VIGS system was optimized for the functional characterization of NbGAE6 homologous genes in N. benthamiana. Whilst the GAE family is well-known for the interconversion of UDP-D-GlcA and UDP-D-GalA during pectin synthesis. Our results revealed that the downregulation of these genes significantly reduced the amount of GalA in the homogalacturunan which is the major component of pectin found in primary cell wall. Biphenyl assay and high performance liquid chromatography analysis (HPLC) depicted that the level of 'GalA' monosaccharide reduced to 40-51% in VIGS plants as compared to the wild type plants. Moreover, qRT-PCR also confirmed the downregulation of the NbGAE6 mRNA in VIGS plants. In all, this is the first comprehensive study of the optimization of VIGS system for the provision of rapid silencing of GAE family members in N. benthamiana, eliminating the need of stable transformants.


Assuntos
Proteínas de Arabidopsis/genética , Carboidratos Epimerases/genética , Parede Celular/metabolismo , Pectinas/genética , Tabaco/genética , Arabidopsis/genética , Parede Celular/genética , Parede Celular/virologia , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Vetores Genéticos/genética , Monossacarídeos/metabolismo , Pectinas/biossíntese , Peptídeos , Vírus de Plantas/genética , RNA Mensageiro/genética , Tabaco/virologia
3.
J Agric Food Chem ; 68(8): 2588-2596, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32031793

RESUMO

Monosaccharides play important roles in plant growth and development, and their biofunctions are closely related to their endogenous contents. Therefore, the determination of monosaccharides is beneficial for the further study of monosaccharide biofunction. In this work, we developed a liquid chromatography-mass spectrometry analytical method assisted by a post-column derivatization technique (LC-PCD-MS) for the fast and automatic determination of 16 monosaccharides in samples. Post-column chemical derivatization of monosaccharides was performed by a reaction of monosaccharides with 4-benzylaminobenzeneboronic acid (4-PAMBA) through boronate ester formation in a three-way connector. 4-PAMBA worked as a derivatization reagent to improve the selectivity and sensitivity of monosaccharide detection by MS. The developed LC-PCD-MS method integrates LC separation, chemical derivatization, and MS detection in one run, thus greatly reducing the analysis time for each sample. The limits of detection and limits of quantification for 16 monosaccharides were in the range of 0.002-0.1 and 0.007-0.5 ng/mL, respectively. Good linearity was obtained from the linear regression, with a determination coefficient (R2) ranging from 0.9928 to 1.0000. The relative recoveries were in the range of 80.7-117.8%, with the intra- and interday relative standard deviations less than 19.7 and 16.5%, respectively, indicating good accuracy and acceptable reproducibility of the method. Finally, the method was successfully applied to investigate the spatial and temporal distribution of 16 monosaccharides in the developing flower and germinating seed of Arabidopsis thaliana.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Monossacarídeos/análise , Espectrometria de Massas em Tandem/métodos , Arabidopsis/química , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Cromatografia Líquida de Alta Pressão/instrumentação , Flores/química , Flores/crescimento & desenvolvimento , Flores/metabolismo , Limite de Detecção , Monossacarídeos/metabolismo , Sementes/química , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Espectrometria de Massas em Tandem/instrumentação
4.
Fish Shellfish Immunol ; 98: 25-33, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31904539

RESUMO

Mannose-binding lectin (MBL) is a crucial pattern recognition receptor in the host innate immune system. Previously, we reported the biological function of Ctenopharyngodon idella MBL (CiMBL) in initiating the lectin pathway of the complement system. In the present study, we further explored its biological function including the agglutinating ability, binding capacity and protective role in vitro and in vivo. After Aeromonas hydrophila infection, western blot analysis revealed that the CiMBL were fluctuated and expressed in the serum and major immune-related tissues. The result of quantitative PCR (qPCR) showed that the recombinant CiMBL (rCiMBL) significantly inhibited the mRNA expression of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in liver, spleen and hepatic cells. Due to rCiMBL bound to d-mannose, d-galactose, d-glucose, N-acetyl-d-glucosamine (GlcNAc), lipopolysaccharide (LPS), peptidoglycan (PGN) and Agar in the presence of Ca2+, herein gram-positive (Staphylococcus aureus and Micrococcus luteus) and gram-negative (A. hydrophila and Vibrio anguillarum) bacteria were agglutinated by rCiMBL in a Ca2+-dependent manner. More importantly, rCiMBL enhanced the survival rate of grass carp following bacterial infection. Overall, the results provide an evidence that CiMBL can protect grass carp against A. hydrophila infection in aquaculture.


Assuntos
Aglutinação , Carpas/imunologia , Doenças dos Peixes/imunologia , Lectina de Ligação a Manose/imunologia , Monossacarídeos/metabolismo , Polissacarídeos/metabolismo , Aeromonas hydrophila/fisiologia , Animais , Carpas/metabolismo , Proteínas de Peixes/imunologia , Proteínas de Peixes/farmacologia , Bactérias Gram-Negativas/fisiologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Bactérias Gram-Positivas/fisiologia , Lectina de Ligação a Manose/farmacologia , Substâncias Protetoras/farmacologia
5.
Food Chem ; 315: 126221, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32000077

RESUMO

Xylo- and arabinoxylo-oligosaccharides (XOS and AXOS) are of interest for their prebiotic activity. The production of these oligomers might be accompanied with monosaccharides. The measurement of both oligosaccharides and monosaccharides usually requires two methods. The current work presents an HPAEC-PAD method based on gradient elution of aqueous solvents sodium hydroxide and sodium acetate, in contrast to conventional isocratic elution, for the simultaneous separation of 16 standards of monosaccharides, xylo-oligosaccharides, arabinoxylo-oligosaccharides and uronic acids using CarboPac PA 200 column. The presented method showed a stable baseline and high-resolution separation of the standards. The method showed acceptable accuracy and precision. Limits of Detection and Quantitation (LOD and LOQ) were estimated for all the standards. The method was applied to measure the activity of a commercial endoxylanase on wheat bran; a steady release of xylose monosaccharide was observed. Enzyme action on oligosaccharide standards showed a preference for the larger oligosaccharides.


Assuntos
Monossacarídeos/análise , Oligossacarídeos/análise , Prebióticos/análise , Triticum/química , Arabinose/análise , Arabinose/metabolismo , Cromatografia por Troca Iônica , Fibras na Dieta , Grão Comestível/química , Endo-1,4-beta-Xilanases/metabolismo , Limite de Detecção , Monossacarídeos/metabolismo , Oligossacarídeos/metabolismo , Reprodutibilidade dos Testes , Ácidos Urônicos/química , Xilose/análise , Xilose/metabolismo
6.
Soft Matter ; 16(5): 1246-1258, 2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-31912078

RESUMO

Simple sugars such as glucose and sucrose are ubiquitous in all organisms. One remarkable property of these small solutes is their ability to protect biomembranes against dehydration damage. This property, which reflects the underlying sugar-lipid interactions, has been intensely studied for lipid bilayers interacting with a single sugar at low hydration. Here, we use giant vesicles to investigate fully hydrated lipid membranes in contact with two sugars, glucose and sucrose. The vesicles were osmotically balanced, with the same total sugar concentration in the interior and exterior aqueous solutions. However, the two solutions differed in their composition: the interior solution contained only sucrose whereas the exterior one contained primarily glucose. This sugar asymmetry generated a striking variety of multispherical or "multi-balloon" vesicle shapes. Each multisphere involved only a single membrane that formed several spherical segments, which were connected by narrow, hourglass-shaped membrane necks. These morphologies revealed that the sugar-lipid interactions generated a significant spontaneous curvature with a magnitude of about 1 µm-1. Such a spontaneous curvature can be generated both by depletion and by adsorption layers of the sugar molecules arising from effectively repulsive and attractive sugar-lipid interactions. All multispherical shapes are stable over a wide range of parameters, with a substantial overlap between the different stability regimes, reflecting the rugged free energy landscape in shape space. One challenge for future studies is to identify pathways within this landscape that allow us to open and close the membrane necks of these shapes in a controlled and reliable manner. We will then be able to apply these multispheres as metamorphic chambers for chemical reactions and nanoparticle growth.


Assuntos
Membrana Celular/metabolismo , Monossacarídeos/metabolismo , Lipossomas Unilamelares/química , Adsorção , Membrana Celular/química , Glucose/análise , Glucose/metabolismo , Lipídeos/química , Monossacarídeos/química , Osmose , Lipossomas Unilamelares/metabolismo
7.
Curr Opin Gastroenterol ; 36(2): 147-154, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31850930

RESUMO

PURPOSE OF REVIEW: Fermentable oligosaccharides disaccharides monosaccharides and polyols (FODMAP) dietary restriction ameliorates irritable bowel syndrome (IBS) symptoms; however, not all individuals with IBS respond. Given the gut microbiome's role in carbohydrate fermentation, investigators have evaluated whether the gut microbiome may predict low FODMAP diet efficacy. RECENT FINDINGS: Gut microbiome fermentation, even to the same carbohydrate, is not uniform across all individuals with several factors (e.g. composition) playing a role. In both children and adults with IBS, studies are emerging suggesting the gut microbiome may predict low FODMAP diet efficacy. However, there is significant heterogeneity in the approaches (study population, microbiome assessment methods, statistical techniques, etc.) used amongst these studies. SUMMARY: The gut microbiome holds promise as a predictor of low FODMAP diet efficacy. However, further investigation using standardized approaches to evaluate the microbiome while concomitantly assessing other potential predictors are needed to more rigorously evaluate this area.


Assuntos
Dieta com Restrição de Carboidratos/métodos , Microbioma Gastrointestinal/fisiologia , Síndrome do Intestino Irritável/dietoterapia , Monossacarídeos/administração & dosagem , Oligossacarídeos/administração & dosagem , Polímeros/administração & dosagem , Dissacarídeos/administração & dosagem , Dissacarídeos/metabolismo , Fermentação , Gastroenteropatias/dietoterapia , Gastroenteropatias/metabolismo , Gastroenteropatias/fisiopatologia , Humanos , Síndrome do Intestino Irritável/metabolismo , Síndrome do Intestino Irritável/fisiopatologia , Monossacarídeos/metabolismo , Oligossacarídeos/metabolismo , Polímeros/metabolismo , Resultado do Tratamento
8.
Food Chem ; 310: 125818, 2020 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31787397

RESUMO

Research on plant biostimulants is of interest in their potential benefits for agriculture production and environmental sustainability. These naturally occurring products induce beneficial consequences in plant metabolism and productivity. In most cases their modes of action, and consequences for the whole plant as well as parts, such as the fruit, are well characterized, but the precise mechanisms of action require further attention. This study examined the effects of the commercial biostimulant, Actium®, on Capsicum annuum L. cv Palermo leaves and fruits. The influence of time (characterized by ripening), after 14 and 28 days of treatment, treatment regimen, and their combined impact on the metabolome were studied using HPLC-ESI-QTOF-MS analysis of polar and apolar compounds. The results showed that flavonoids and capsianosides decreased with ripening in leaves, but organic acids, monosaccharides, and carotenoids increased in fruits. The treatment of Capsicum fruits with Actium® increased phenylalanine and total monosaccharides (glucose and fructose) compared to controls, suggesting a further stage in ripening. An increase in carotenoids concomitant with an increase of some digalactosyl diacylglycerols, which are part of the chromoplasts lipid machinery of enzymes involved in the synthesis of carotenoids, was also observed. Our findings suggest that this biostimulant may increase some metabolites related to pepper fruit maturity and coloration in pepper crops.


Assuntos
Capsicum/metabolismo , Frutas/metabolismo , Metaboloma , Reguladores de Crescimento de Planta/farmacologia , Folhas de Planta/metabolismo , Capsicum/efeitos dos fármacos , Carotenoides/metabolismo , Cromatografia Líquida de Alta Pressão , Produtos Agrícolas/metabolismo , Frutas/efeitos dos fármacos , Frutas/fisiologia , Metabolômica , Monossacarídeos/metabolismo , Fenilalanina/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/fisiologia , Espectrometria de Massas por Ionização por Electrospray
9.
J. pediatr. (Rio J.) ; 95(6): 642-656, Nov.-Dec. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1056660

RESUMO

ABSTRACT Objective: This narrative review aimed to provide practitioners a synthesis of the current knowledge on the role of a low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet in reducing symptoms associated with functional abdominal pain disorders in children. This review is focused on the pathophysiology, efficacy and criticism of low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet in children. Sources: Cochrane Database, Pubmed and Embase were searched using specific terms for Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet interventions and functional abdominal pain disorders. Summary of the findings: In children, only one Randomized Control Trial and one open-label study reported positive results of low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet; one Randomized Control Trial showed exacerbation of symptoms with fructans in children with Irritable Bowel Syndrome; no effect was found for the lactose-free diet whilst fructose-restricted diets were effective in 5/6 studies. Conclusions: In children there are few trials evaluating low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols in functional abdominal pain disorders, with encouraging data on the therapeutic efficacy particularly of fructose-restricted diet. Additional efforts are still needed to fill this research gap and clarify the most efficient way for tailoring dietary restrictions based on the patient's tolerance and/or identification of potential biomarkers of low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols efficacy, to maintain nutritional adequacy and to simplify the adherence to diet by labeling Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols content in commercial products.


RESUMO Objetivo: Nos últimos anos, foram feitos esforços consideráveis para esclarecer o papel da dieta com baixo teor de oligossacarídeos fermentáveis, dissacarídeos, monossacarídeos e polióis (FODMAPs) para o tratamento de distúrbios gastrintestinais funcionais (DGIFs). Esta revisão narrativa teve como objetivo fornecer aos profissionais uma síntese do conhecimento atual sobre o papel de uma dieta com baixo teor de FODMAPs (BFM) na redução dos sintomas associados a distúrbios funcionais de dor abdominal (DFDA) em crianças. Esta revisão está focada na fisiopatologia, eficácia e crítica da dieta BFM em crianças. Fontes: O banco de dados Cochrane, Pubmed e Embase foram pesquisados com o uso dos termos específicos para intervenções na dieta FODMAP e DFDA. Resumo dos achados: Em crianças, apenas um estudo controlado randomizado e um estudo aberto relataram resultados positivos da dieta BFM; um estudo controlado randomizado mostrou exacerbação dos sintomas com frutanos em crianças com síndrome do intestino irritável; nenhum efeito foi encontrado para a dieta livre de lactose, enquanto dietas com restrição de frutose foram eficazes em 5/6 estudos. Conclusões: Existem poucos estudos que avaliam BFM em DFDA em crianças, com dados encorajadores sobre a eficácia terapêutica, particularmente de dietas com restrição de frutose. Esforços adicionais ainda são necessários para preencher essa lacuna de pesquisa e esclarecer a maneira mais eficiente de adaptar as restrições dietéticas com base na tolerância do paciente e/ou identificação de biomarcadores potenciais de eficácia da BFM, para manter a adequação nutricional e simplificar a adesão à dieta, ao incluir informações sobre conteúdo de FODMAPs em rótulos de produtos comerciais.


Assuntos
Humanos , Dor Abdominal/dietoterapia , Dieta com Restrição de Carboidratos , Oligossacarídeos/metabolismo , Oligossacarídeos/uso terapêutico , Síndrome do Intestino Irritável , Dieta , Dissacarídeos/metabolismo , Dissacarídeos/uso terapêutico , Monossacarídeos/metabolismo , Monossacarídeos/uso terapêutico
10.
Nat Commun ; 10(1): 4065, 2019 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-31492838

RESUMO

Unnatural monosaccharides such as azidosugars that can be metabolically incorporated into cellular glycans are currently used as a major tool for glycan imaging and glycoproteomic profiling. As a common practice to enhance membrane permeability and cellular uptake, the unnatural sugars are per-O-acetylated, which, however, can induce a long-overlooked side reaction, non-enzymatic S-glycosylation. Herein, we develop 1,3-di-esterified N-azidoacetylgalactosamine (GalNAz) as next-generation chemical reporters for metabolic glycan labeling. Both 1,3-di-O-acetylated GalNAz (1,3-Ac2GalNAz) and 1,3-di-O-propionylated GalNAz (1,3-Pr2GalNAz) exhibit high efficiency for labeling protein O-GlcNAcylation with no artificial S-glycosylation. Applying 1,3-Pr2GalNAz in mouse embryonic stem cells (mESCs), we identify ESRRB, a critical transcription factor for pluripotency, as an O-GlcNAcylated protein. We show that ESRRB O-GlcNAcylation is important for mESC self-renewal and pluripotency. Mechanistically, ESRRB is O-GlcNAcylated by O-GlcNAc transferase at serine 25, which stabilizes ESRRB, promotes its transcription activity and facilitates its interactions with two master pluripotency regulators, OCT4 and NANOG.


Assuntos
Acetilglucosamina/metabolismo , Monossacarídeos/metabolismo , Células-Tronco Embrionárias Murinas/metabolismo , Células-Tronco Pluripotentes/metabolismo , Receptores Estrogênicos/metabolismo , Animais , Azidas/química , Azidas/metabolismo , Linhagem Celular Tumoral , Autorrenovação Celular , Células Cultivadas , Glicosilação , Células HeLa , Hexosaminas/metabolismo , Humanos , Células MCF-7 , Camundongos , Monossacarídeos/química , Células-Tronco Embrionárias Murinas/citologia , Células NIH 3T3 , Células-Tronco Pluripotentes/citologia , Processamento de Proteína Pós-Traducional
11.
Int J Biol Macromol ; 141: 307-312, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31491510

RESUMO

In this study, we demonstrate that chitosan is produced from Paecilomyces saturatus fungi using ternary monosaccharide carbon sources liquid cultivation via mixture design strategy. Sixteen experiments were carried out to obtain regression equations of fungal dry mycelial biomass (W), chitosan ratio (R), and deacetylation degree (DD) for plotting contour lines. Contour lines reveal that the maximum W, R, and DD can be simultaneously obtained in cultivated media containing 20% glucose, 60% fructose and 20% mannitol rather than pure monosaccharide cultivation. Three additional confirmation experiments based on the maximum FuCS deacetylation degree had been performed to confirm to be 92.3% via Fourier-transform infrared spectra. Accordingly, FuCS possessed much better anti-microbial activity on E. coli than commercial chitosan (CrCS). Meanwhile, X-ray diffraction results confirmed that FuCS possessed both α and γ crystalline peaks while CrCS possessed only α crystalline peak, being collaborated with thermogravimetric analysis results. The superior FuCS was obtained by using ternary monosaccharides system in fungal culture via mixture design for the first time. This study provides a new approach to produce chitosan from fungal cultivation by using the mixture design strategy.


Assuntos
Quitosana/metabolismo , Engenharia Metabólica , Monossacarídeos/metabolismo , Paecilomyces/metabolismo , Acetilação , Biomassa , Quitosana/análise , Quitosana/isolamento & purificação , Engenharia Metabólica/métodos , Análise Espectral
12.
Med Hypotheses ; 132: 109324, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31421429

RESUMO

Functional bowel symptoms can be occurred during remission from inflammatory bowel disease. In this case, a low fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAP) diet is effective for the amelioration or prevention of symptoms. However, the reason is not fully explained. This report proposes a hypothesis regarding the entire process in which inflammatory bowel disease with IBS-like symptoms (IBD-IBS) causes symptoms. A detailed process was assumed, starting from high pressure in the lumen and finally to abdominal symptoms. In this process, relationships were linked based on interactions such as ischemia, compliance, pain threshold, visceral hypersensitivity, mast cells, and permeability reported in IBD-IBS. In the process mapping, to understand the relationship between the amount of gas increased by FODMAP and ischemia, the hydrodynamic hypothesis and Ritchie's hypothesis were adapted. Ischemia in dilated intestines due to an increase in gas volume can induce excessive spasms via the mast cells and show the whole process of lowering the pain threshold. From the standpoint of the mechanism of IBD-IBS, the origin trigger may be FODMAP. Therefore, a low-FODMAP diet is recommended to relieve and prevent IBD-IBS symptoms.


Assuntos
Doenças Inflamatórias Intestinais/fisiopatologia , Síndrome do Intestino Irritável/fisiopatologia , Dieta , Dissacarídeos/metabolismo , Fermentação , Humanos , Sulfeto de Hidrogênio/química , Concentração de Íons de Hidrogênio , Intestinos , Isquemia , Mastócitos/metabolismo , Monossacarídeos/metabolismo , Oligossacarídeos/metabolismo , Permeabilidade , Polímeros/metabolismo , Pressão , Qualidade de Vida , Canais de Cátion TRPV/metabolismo
13.
Anal Chem ; 91(17): 11070-11077, 2019 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-31407893

RESUMO

"The totality is not, as it were, a mere heap, but the whole is something besides the parts."-Aristotle. We built a classifier that uses the totality of the glycomic profile, not restricted to a few glycoforms, to differentiate samples from two different sources. This approach, which relies on using thousands of features, is a radical departure from current strategies, where most of the glycomic profile is ignored in favor of selecting a few features, or even a single feature, meant to capture the differences in sample types. The classifier can be used to differentiate the source of the material; applicable sources may be different species of animals, different protein production methods, or, most importantly, different biological states (disease vs healthy). The classifier can be used on glycomic data in any form, including derivatized monosaccharides, intact glycans, or glycopeptides. It takes advantage of the fact that changing the source material can cause a change in the glycomic profile in many subtle ways: some glycoforms can be upregulated, some downregulated, some may appear unchanged, yet their proportion-with respect to other forms present-can be altered to a detectable degree. By classifying samples using the entirety of their glycan abundances, along with the glycans' relative proportions to each other, the "Aristotle Classifier" is more effective at capturing the underlying trends than standard classification procedures used in glycomics, including PCA (principal components analysis). It also outperforms workflows where a single, representative glycomic-based biomarker is used to classify samples. We describe the Aristotle Classifier and provide several examples of its utility for biomarker studies and other classification problems using glycomic data from several sources.


Assuntos
Glicômica/métodos , Glicopeptídeos/classificação , Glicoproteínas/classificação , Cirrose Hepática/diagnóstico , Monossacarídeos/classificação , Polissacarídeos/classificação , Biomarcadores/análise , Glicopeptídeos/isolamento & purificação , Glicopeptídeos/metabolismo , Glicoproteínas/isolamento & purificação , Glicoproteínas/metabolismo , Glicosilação , Humanos , Cirrose Hepática/metabolismo , Monossacarídeos/isolamento & purificação , Monossacarídeos/metabolismo , Polissacarídeos/isolamento & purificação , Polissacarídeos/metabolismo , Análise de Componente Principal , Software , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Terminologia como Assunto
14.
Biotechnol Lett ; 41(10): 1187-1200, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31418101

RESUMO

OBJECTIVES: Bifunctional alginate lyase can efficiently saccharify alginate biomass and prepare functional oligosaccharides of alginate. RESULTS: A new BP-2 strain that produces alginate lyase was screened and identified from rotted Sargassum. A new alginate lyase, Alg17B, belonging to the polysaccharide lyase family 17, was isolated and purified from BP-2 fermentation broth by freeze-drying, dialysis, and ion exchange chromatography. The enzymatic properties of the purified lyase were investigated. The molecular weight of Alg17B was approximately 77 kDa, its optimum reaction temperature was 40-45 °C, and its optimum reaction pH was 7.5-8.0. The enzyme was relatively stable at pH 7.0-8.0, with a temperature range of 25-35 °C, and the specific activity of the purified enzyme reached 4036 U/mg. A low Na+ concentration stimulated Alg17B enzyme activity, but Ca2+, Zn2+, and other metal ions inhibited it. Substrate specificity analysis, thin-layer chromatography, and mass spectrometry showed that Alg17B is an alginate lyase that catalyses the hydrolysis of sodium alginate, polymannuronic acid (polyM) and polyguluronic acid to produce monosaccharides and low molecular weight oligosaccharides. Alg17B is also bifunctional, exhibiting both endolytic and exolytic activities toward alginate, and has a wide substrate utilization range with a preference for polyM. CONCLUSIONS: Alg17B can be used to saccharify the main carbohydrate, alginate, in the ethanolic production of brown algae fuel as well as in preparing and researching oligosaccharides.


Assuntos
Organismos Aquáticos/enzimologia , Gammaproteobacteria/enzimologia , Polissacarídeo-Liase/isolamento & purificação , Polissacarídeo-Liase/metabolismo , Sargassum/microbiologia , Alginatos/metabolismo , Ácido Algínico/metabolismo , Organismos Aquáticos/classificação , Organismos Aquáticos/genética , Organismos Aquáticos/isolamento & purificação , Ativadores de Enzimas/análise , Inibidores Enzimáticos/análise , Estabilidade Enzimática , Gammaproteobacteria/classificação , Gammaproteobacteria/genética , Gammaproteobacteria/isolamento & purificação , Concentração de Íons de Hidrogênio , Hidrólise , Peso Molecular , Monossacarídeos/metabolismo , Oligossacarídeos/metabolismo , Polissacarídeo-Liase/química , Polissacarídeo-Liase/genética , Polissacarídeos Bacterianos/metabolismo , Especificidade por Substrato , Temperatura
15.
Clin Nutr ESPEN ; 33: 18-28, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31451258

RESUMO

BACKGROUND AND AIMS: Monosaccharides are important components of the diet, where the sweetness of these common sugars draw animals to eat the tissue within which they are located - especially fruits. Higher (larger) saccharides, within which they are constituents, are ubiquitous throughout nature too - and include disaccharides, oligosaccharides and polysaccharides. These may be converted (hydrolysed) to monosaccharides by the plant tissue enzymes during ripening and stimulate consumption by a predator (whereupon seeds within the fruit are dispersed). Predators may have relevant enzymes in their digestive tract to effect conversion of the larger carbohydrates to its monosaccharides - which are then absorbed from the gut and like free monosaccharides in fruit/vegetables, provide an energy source. Starch is an important source of glucose. This review (on monosaccharides) is part one of a series of three which aim to link the role of carbohydrates in food through processing to health and disease related issues. The emphasis here is to understand the role of the three key monosaccharides from the diet - fructose, galactose and glucose - with perspectives in health and disease. METHODS: The review was based on a review of relevant databases for material (e.g. Pubmed, Science Direct, Web of Science, Wiley online library etc.). RESULTS: Data pertaining to the nutritional role of key dietary monosaccharides were evaluated together with their utilisation and role in health and disease. Disease states and their management in the context of monosaccharide consumption were considered. CONCLUSIONS: The body is designed to utilise carbohydrates - where a physiological balance of ingestion, storage and utilisation is critical. In disease states, the balance is lost and a number of carbohydrate based metabolic disorders are established within the medical community. Overall, this review considers digestive and metabolic issues associated with free monosaccharides commonly consumed in the human diet. Further reviews will focus on common di-, oligo and polysaccharides relevant to digestive energy and overall health.


Assuntos
Carboidratos da Dieta/metabolismo , Doença , Frutose/metabolismo , Galactose/metabolismo , Glucose/metabolismo , Saúde , Animais , Dieta , Dissacarídeos/metabolismo , Humanos , Erros Inatos do Metabolismo , Monossacarídeos/metabolismo , Oligossacarídeos/metabolismo , Polissacarídeos/metabolismo , Amido/metabolismo
16.
Int J Mol Sci ; 20(16)2019 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-31426432

RESUMO

Sugar transporters of the SWEET family mediate cross membrane movement of mono- and disaccharides and play vital roles in diverse physiological and pathophysiological processes, including sink-source relationship, pathogen responses, reproductive growth, and development. However, it remains to be determined how these transporters function in non-module plants of agricultural significance, given the evolutionarily diverse traits. In this study, we combined transcriptome analysis, rapid amplification of cDNA ends-cloning (RACE-cloning), expression profiling, and heterologous functional assay to identify SWEET genes that may have potential roles during flower opening and sexual reproduction in Jasminum sambac . During the anthesis, the floral organs of J. sambac express seven SWEET homologous genes from all four clades of the family. JsSWEET9 and 2 are significantly upregulated when flowers are fully opened, up to 6- and 3-fold compared to unopened buds, respectively. The other transporters, JsSWEET1, 5, 10, and 17 are also accumulated slightly at stage associated with fragrance release, whereas only the vacuole transporter JsSWEET16 showed small decrease in transcript level after anthesis. The JsSWEET5, a clade II member, is capable to complement yeast cell uptake on most tested sugar substrates with a preference for hexoses, while the clade I transporter JsSWEET1 mediates merely galactose import when expressed in yeast. Our results provide first evidence for further investigation on sugar transport and allocation during flowering and reproductive processes in J. sambac.


Assuntos
Flores/genética , Jasminum/genética , Proteínas de Membrana Transportadoras/genética , Proteínas de Plantas/genética , Clonagem Molecular , Dissacarídeos/metabolismo , Flores/crescimento & desenvolvimento , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Jasminum/crescimento & desenvolvimento , Jasminum/metabolismo , Proteínas de Membrana Transportadoras/análise , Proteínas de Membrana Transportadoras/metabolismo , Proteínas de Transporte de Monossacarídeos/análise , Proteínas de Transporte de Monossacarídeos/genética , Proteínas de Transporte de Monossacarídeos/metabolismo , Monossacarídeos/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/metabolismo
17.
Thromb Haemost ; 119(8): 1265-1273, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31266083

RESUMO

Complex glycans are readily accessible on the endothelium and on cell and plasma components. They interact with glycan-binding proteins which translate their structure into function. Advanced analytical tools are available to investigate their structure and functional interactions. Modifications to glycan structures which alter their capacity to bind proteins are particularly relevant in atherosclerosis. We summarize the regulatory role of glycans and their binding partners in the development of the disease. Given their complexity, accessibility, and important functional role, glycans and glycan-binding proteins represent promising diagnostic tools and therapeutic targets.


Assuntos
Aterosclerose/metabolismo , Proteínas de Transporte/metabolismo , Glicocálix/metabolismo , Polissacarídeos/metabolismo , Animais , Adesão Celular , Movimento Celular , Galectinas/metabolismo , Humanos , Lectinas/metabolismo , Lectinas Tipo C/metabolismo , Leucócitos/citologia , Metabolismo dos Lipídeos , Lectina de Ligação a Manose/metabolismo , Camundongos , Monossacarídeos/metabolismo , Ligação Proteica , Receptores de LDL/metabolismo , Transdução de Sinais , Trombose/metabolismo , Resultado do Tratamento
18.
PLoS One ; 14(6): e0217679, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31216303

RESUMO

Ruditapes philippinarum conglutination mud (RPM) is a byproduct from the aquiculture of an important commercially bivalve mollusk R. philippinarum and has been recently reported as a promising natural bioflocculant resource. However the origin of bioflocculation components within RPM is still a pending doubt and impedes its effective exploitation. This study investigated the probability that RPM bioflocculation components originate from its associated microbes. RPM samples from an aquaculture farm in Zhoushan of China were applied to characterize its microbial community structure, screen associated bioflocculant-producing strains, and explore the homology between extracellular polysaccharides (EPS) from bioflocculant-producing isolates and RPM flocculation components. Results showed that RPM exhibited high bacterial biodiversity, with Proteobacteria, Bacteroidetes and Actinobacteria as the most abundant phyla; hgcI_clade, CL500_29_marine_group, Fusibacter, MWH_UniP1_aquatic_group and Arcobacter as the dominant genera. Fourteen highly efficient bioflocculant-producing strains were screened and phylogenetically identified as Pseudoalteromonas sp. (5), Psychrobacter sp. (3), Halomonas sp. (2), Albirhodobacter sp. (1), Celeribacter sp. (1), Kocuria sp. (1) and Bacillus sp. (1), all of which except Bacillus sp. were reported for the first time for their excellent flocculation capability. Furthermore, EPS from the bioflocculant-producing strains exhibited highly similar monosaccharide composition to the reported flocculation-effective RPM polysaccharides. On the other hand, the existence of fungi in RPM was rare and showed no flocculation functionality. Findings from Zhoushan RPM strongly supported that RPM flocculation components were of bacterial origin and make RPM reproduction possible by fermentation approach.


Assuntos
Bactérias/genética , Bivalves/microbiologia , Recursos Naturais , Animais , Aquicultura , Bactérias/metabolismo , Bivalves/metabolismo , China , Fazendas , Floculação , Fungos/genética , Fungos/metabolismo , Humanos , Monossacarídeos/genética , Monossacarídeos/metabolismo , Reação em Cadeia da Polimerase , Polissacarídeos/genética , Polissacarídeos/metabolismo , Água do Mar/microbiologia , Temperatura
19.
PLoS One ; 14(5): e0217435, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31120985

RESUMO

Sudangrass, Sorghum sudanense (Piper) Stapf, is a vigorous forage crop that has also been used for biogas, paper, and electricity production. Due to the large biomass yields achieved by sudangrass and the large area of potential growth in Argentina seven sudangrass accessions from a collection of S. sudanense were analyzed to evaluate their potential as feedstocks for lignocellulosic bioethanol production, and to assess whether there is an association between the response to biotic and abiotic stresses and the composition of the biomass. The biomass composition was analyzed for major cell wall polymers, monosaccharides, and elemental composition. On average, 68% of stem lignocellulosic biomass was comprised of matrix polysaccharides and crystalline cellulose, representing a potential source of sugars for bioethanol production. Xylose was the predominant matrix polysaccharide monosaccharide comprising, on average, 45% of the total sugars, followed by arabinose, glucose, galactose, galacturonic acid, mannose, glucuronic acid, and fucose. Rhamnose was not detected in any of the biomasses analyzed. Silica was the most abundant element in sudangrass stem, followed by chloride, calcium, phosphorus and sulfur. We performed saccharification analyses after pretreatments. Alkaline pretreatment was more effective than water pretreatment. Sodium hydroxide pretreatment exposed different levels of recalcitrance among sudangrass accessions, whereas the water pretreatment did not. Phenological traits were also evaluated, showing significant variability among accessions. The comparison of major cell wall polymers and monosaccharide composition between tolerant and susceptible accessions to abiotic and biotic stresses suggests an association between the composition of the biomass and the response to stress.


Assuntos
Fontes de Energia Bioelétrica , Biomassa , Etanol/metabolismo , Lignina/metabolismo , Sorghum/fisiologia , Argentina , Parede Celular/química , Parede Celular/metabolismo , Lignina/análise , Monossacarídeos/análise , Monossacarídeos/metabolismo , Polissacarídeos/análise , Polissacarídeos/metabolismo , Sorghum/química , Estresse Fisiológico
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