The carbohydrate-active enzyme database: functions and literature.

Thirty years have elapsed since the emergence of the classification of carbohydrate-active enzymes in sequence-based families that became the CAZy database over 20 years ago, freely available for browsing and download at www.cazy.org. In the era of large scale sequencing and high-throughput Biology, it is important to examine the position of this specialist database that is deeply rooted in human curation. The three primary tasks of the CAZy curators are (i) to maintain and update the family classification of this class of enzymes, (ii) to classify sequences newly released by GenBank and the Protein Data Bank and (iii) to capture and present functional information for each family. The CAZy website is updated once a month. Here we briefly summarize the increase in novel families and the annotations conducted during the last 8 years. We present several important changes that facilitate taxonomic navigation, and allow to download the entirety of the annotations. Most importantly we highlight the considerable amount of work that accompanies the analysis and report of biochemical data from the literature.

Drosophila insulin-like peptides regulate concentration-dependent changes of appetite to different carbohydrates.

The volumes of sugar solutions ingested and amounts of different carbohydrates eaten were measured in fruit fly lines with mutated genes for Drosophila insulin-like peptides (DILPs). The wild type w1118 flies consumed 20-40 µg of fructose or glucose per day regardless of carbohydrate concentration. This relatively constant amount of consumed carbohydrate was regulated due to satiety-driven decreases in the ingested volume of sugar solution, a so-called "compensatory feeding" strategy. This decrease was not observed for flies fed sucrose solutions. The dilp3 mutant and quadruple mutant dilp1-4 showed no "compensatory feeding" when fed glucose but these two mutants consumed larger amounts of sucrose than the wild type from solutions with carbohydrate concentrations equal to or higher than 4%. Flies with mutations of dilp2, dilp3, dilp4, dilp5, and dilp6 genes consumed larger amounts of carbohydrate from 4-10% sucrose solutions as compared to the wild type. Mutations of DILPs affected appetite mainly for sucrose and glucose, but the least for fructose. The presented data confirm our hypothesis that DILPs are involved in the regulation of fly appetite in response to type and concentration of carbohydrate.

Influence of Partial Acid Hydrolysis on Size, Dispersity, Monosaccharide Composition, and Conformation of Linearly-Branched Water-Soluble Polysaccharides.

The effect of partial acid hydrolysis on the physical and chemical properties of galactomannan, arabinoxylan, and xyloglucan was investigated. Polysaccharides were treated at 50 °C with hydrochloric acid for 3-48 h. Portions of isopropanol (i-PrOH) were added sequentially to the hydrolyzates, resulting in fractions that were collected by centrifugation. As expected, a significant reduction of weight-average molecular weight (Mw) was observed with increasing hydrolysis time. Fractional precipitation was successfully applied to collect at least one polymer fraction with dispersity (D) close to one for each polysaccharide. The monosaccharide composition analysis showed that the partial hydrolysis usually lowered the relative amount of side chains, with the exception of galactomannan, where the composition remained largely unaffected. Estimation of the polymer conformation in solution, through evaluation of the Mark-Houwink parameter coefficient (α), confirmed that acid hydrolysis influenced the polysaccharides' conformation. It was demonstrated that acid treatment in dilute solution followed by fractional isopropanol precipitation is a method, extendible to a variety of polysaccharides, to obtain materials of decreased molecular weight and low dispersity with slightly altered overall composition and conformation.

Microbial protein formation of different carbohydrates in vitro.

The aim of this study was to investigate the microbial protein yield of different pure carbohydrates to contribute to a more precise prediction of the microbial protein formed in the rumen. In a first experiment, sucrose, wheat starch, microcrystalline cellulose and citrus pectin were incubated for 8 and 24 hr in the modified Hohenheim gas test (HGT) system (3 runs × 2 syringes) including gas production, ammonia and short-chain fatty acid concentration measurements. Ammonia values were used for estimation of the microbial protein formation. In a second experiment, the same substrates were incubated for 96 hr in the HGT system (2 runs × 3 syringes) and gas production was measured after 2, 4, 6, 8, 12, 16, 24, 30, 36, 48, 60, 72 and 96 hr of incubation to obtain the fermentation kinetics and the time of half-maximal gas production (t1/2 ) of the substrates. The substrates differed considerably in their fermentation kinetics, and therefore, comparison on the basis of t1/2 was chosen as the most meaningful. At t1/2 , microbial protein yield [g/kg dry matter] was higher for cellulose than for sucrose and pectin and higher for starch than for sucrose. The microbial protein expressed in g/L gas production was higher for starch and cellulose than for sucrose and pectin at t1/2 . Effects of carbohydrates related to ruminal pH may remain undetected in in vitro trials.

Nondigestible carbohydrates, butyrate, and butyrate-producing bacteria.

Nondigestible carbohydrates (NDCs) are fermentation substrates in the colon after escaping digestion in the upper gastrointestinal tract. Among NDCs, resistant starch is not hydrolyzed by pancreatic amylases but can be degraded by enzymes produced by large intestinal bacteria, including clostridia, bacteroides, and bifidobacteria. Nonstarch polysaccharides, such as pectin, guar gum, alginate, arabinoxylan, and inulin fructans, and nondigestible oligosaccharides and their derivatives, can also be fermented by beneficial bacteria in the large intestine. Butyrate is one of the most important metabolites produced through gastrointestinal microbial fermentation and functions as a major energy source for colonocytes by directly affecting the growth and differentiation of colonocytes. Moreover, butyrate has various physiological effects, including enhancement of intestinal barrier function and mucosal immunity. In this review, several representative NDCs are introduced, and their chemical components, structures, and physiological functions, including promotion of the proliferation of butyrate-producing bacteria and enhancement of butyrate production, are discussed. We also describe the strategies for achieving directional accumulation of colonic butyrate based on endogenous generation mechanisms.

Evolution of a Vegetarian Vibrio: Metabolic Specialization of Vibrio breoganii to Macroalgal Substrates.

While most Vibrionaceae are considered generalists that thrive on diverse substrates, including animal-derived material, we show that Vibrio breoganii has specialized for the consumption of marine macroalga-derived substrates. Genomic and physiological comparisons of V. breoganii with other Vibrionaceae isolates revealed the ability to degrade alginate, laminarin, and additional glycans present in algal cell walls. Moreover, the widely conserved ability to hydrolyze animal-derived polymers, including chitin and glycogen, was lost, along with the ability to efficiently grow on a variety of amino acids. Ecological data showing associations with particulate algal material but not zooplankton further support this shift in niche preference, and the loss of motility appears to reflect a sessile macroalga-associated lifestyle. Together, these findings indicate that algal polysaccharides have become a major source of carbon and energy in V. breoganii, and these ecophysiological adaptations may facilitate transient commensal associations with marine invertebrates that feed on algae.IMPORTANCE Vibrios are often considered animal specialists or generalists. Here, we show that Vibrio breoganii has undergone massive genomic changes to become specialized on algal carbohydrates. Accompanying genomic changes include massive gene import and loss. These vibrios may help us better understand how algal biomass is degraded in the environment and may serve as a blueprint on how to optimize the conversion of algae to biofuels.

Detection, purification and characterization of a lectin from freshwater green algae Spirogyra spp.

Freshwater algae are rich sources of structurally biologically active metabolites, such as fatty acids, steroids, carotenoids and polysaccharides. Among these metabolites, lectins stand out. Lectins are proteins or glycoproteins of non-immune origin which bind to carbohydrates or glycoconjugates, without changing ligand structure. Many studies have reported on the use of Spirogyra spp. as effective bioindicators of heavy metals; however, reports on Spirogyra molecular bioprospecting are quite limited. Therefore, this study aimed to detect, isolate, purify and characterize a lectin present in the freshwater green algae Spirogyra. Presence of the lectin protein in the extract was detected by hemagglutination assays. Subsequently, the protein extract was subjected to a sugar inhibition assay to identify the lectin-specific carbohydrate. Following this, the extract was applied to a guar gum column to afford the pure lectin. The lectin was inhibited by N-acetyl-glucosamine and N-acetyl-beta-D-mannose, but more strongly by D-galactose. The apparent molecular mass of the purified lectin was evaluated by Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Electrophoretic analysis revealed a single protein band with an apparent molecular mass of 56 kDa. Thus, it could be concluded that a lectin was purified from Spirogyra spp.

DrawGlycan-SNFG: a robust tool to render glycans and glycopeptides with fragmentation information.

Glycan or carbohydrate structures can be pictorially represented using symbolic nomenclatures. The symbol nomenclature for glycans (SNFG) contains 67 different monosaccharides represented using various colors and geometric shapes. A simple tool to convert International Union of Pure and Applied Chemistry (IUPAC) format text to SNFG will be useful for sketching glycans and glycopeptides. Such code can also enable the development of more sophisticated applications, where the visual representation of carbohydrate structures is necessary. To address this need, the current manuscript describes DrawGlycan-SNFG, a freely available, platform-independent, open-source tool. It allows: i. the display of glycans and glycopeptides from IUPAC-condensed text inputs and ii. the depiction of glycan and glycopeptide fragments. The online version of this program is provided with a user-friendly web interface at www.virtualglycome.org/DrawGlycan. Downloadable, stand-alone GUI (Graphical User Interface) version and the program source code are also available from this repository. DrawGlycan-SNFG will be useful for experimentalists looking for a ready to use, simple program for sketching carbohydrates and for software developers interested in incorporating SNFG into their program suite.

Detection, purification and characterization of a lectin from freshwater green algae Spirogyra spp

ABSTRACT Freshwater algae are rich sources of structurally biologically active metabolites, such as fatty acids, steroids, carotenoids and polysaccharides. Among these metabolites, lectins stand out. Lectins are proteins or glycoproteins of non-immune origin which bind to carbohydrates or glycoconjugates, without changing ligand structure. Many studies have reported on the use of Spirogyra spp. as effective bioindicators of heavy metals; however, reports on Spirogyra molecular bioprospecting are quite limited. Therefore, this study aimed to detect, isolate, purify and characterize a lectin present in the freshwater green algae Spirogyra. Presence of the lectin protein in the extract was detected by hemagglutination assays. Subsequently, the protein extract was subjected to a sugar inhibition assay to identify the lectin-specific carbohydrate. Following this, the extract was applied to a guar gum column to afford the pure lectin. The lectin was inhibited by N-acetyl-glucosamine and N-acetyl-beta-D-mannose, but more strongly by D-galactose. The apparent molecular mass of the purified lectin was evaluated by Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Electrophoretic analysis revealed a single protein band with an apparent molecular mass of 56 kDa. Thus, it could be concluded that a lectin was purified from Spirogyra spp.

Rethinking the Hierarchy of Sugar Utilization in Bacteria.

Bacteria are known to consume some sugars over others, although recent work reported by Koirala and colleagues in this issue of the Journal of Bacteriology (S. Koirala, X. Wang, and C. V. Rao, J Bacteriol 198:386-393, 2016, http://dx.doi.org/10.1128/JB.00709-15) revealed that individual cells do not necessarily follow this hierarchy. By studying the preferential consumption of l-arabinose over d-xylose in Escherichia coli, those authors found that subpopulations consume one, the other, or both sugars through cross-repression between utilization pathways. Their findings challenge classic assertions about established hierarchies and can guide efforts to engineer the simultaneous utilization of multiple sugars.

Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. glycolytica.

UNLABELLED: Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. The status of such systems in distantly related commensal species, however, remains largely unexplored. Using a strain of deeply branching Neisseria elongata subsp. glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. IMPORTANCE: Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. Here, we examined protein glycosylation in a distantly related, nonpathogenic neisserial species, Neisseria elongata subsp. glycolytica. This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria.

Flavor preference conditioning by different sugars in sweet ageusic Trpm5 knockout mice.

Knockout (KO) mice missing the taste signaling protein Trpm5 have greatly attenuated sweetener preferences but develop strong preferences for glucose in 24-h tests, which is attributed to post-oral sugar conditioning. Trpm5 KO mice express mild preferences for galactose but no preferences for fructose in 24-h tests, which suggests that these sugars differ in their post-oral reinforcing effects. Here we investigated sugar-conditioned flavor preferences in Trpm5 KO and C57BL/6J wildtype (B6) mice. The mice were trained to consume a flavored (CS+, e.g. grape) 8% sugar solution and flavored (CS-, e.g., cherry) water on alternating days followed by two-bottle choice tests with CS+ vs. CS- flavors in water and with unflavored sugar vs. water. The KO mice displayed strong preferences (>80%) for the CS+ glucose and CS+ galactose but not for the CS+ fructose flavor. They also preferred glucose and galactose, but not fructose to water. In contrast, the B6 mice preferred all three CS+ flavors to the CS- flavor, and all three sugars to water. In tests with the non-metabolizable sugar α-methyl-d-glucopyranoside (MDG), the KO and B6 mice preferred 8% MDG to water but did not prefer the CS+ 8% MDG to CS-. However, they preferred a CS+ flavor mixed with 4% MDG over the CS- flavor. Trpm5 KO mice also preferred galactose and MDG to fructose in direct choice tests. The Trpm5 KO data indicate that glucose and, to a lesser extent, galactose and MDG have post-oral reinforcing actions that stimulate intake and preference while fructose has a much weaker effect. The CS+ flavor and sugar preferences of B6 mice may be mediated by the sweet taste and/or post-oral actions of the various sugars. Glucose, galactose, and MDG, but not fructose, are ligands for the sodium-glucose transporter 1 (SGLT1) which is implicated in post-oral sugar conditioning in B6 mice.

Los alimentos como fuente de mono y disacáridos: Aspectos bioquímicos y metabólicos / Food as sources of mono and disaccharides: Biochemical and metabolic aspects

Los hidratos de carbono constituyen una parte importante y necesaria en la alimentación humana. Aunque desempeñan una función primordialmente energética, también tienen funciones estructurales y funcionales. Según la Agencia Europea para la Seguridad Alimentaria, la ingesta de hidratos de carbono debe oscilar entre un 45 y 60% de la energía en adultos y niños mayores de un año. Una parte importante de los hidratos de carbono disponibles en los alimentos la componen los mono y disacáridos, comúnmente denominados azúcares. Las principales fuentes dietéticas de azúcares son las frutas, los zumos de fruta, algunos productos vegetales, la leche y los productos lácteos, y los alimentos que contengan sacarosa añadida e hidrolizados de almidón. A pesar de ser fundamentales en nuestra vida diaria, no existe una terminología adecuada y clara sobre los diversos tipos de hidratos de carbono, y de forma muy especial de los azúcares. Tampoco en lo referente a las recomendaciones de ingesta y contenido en los alimentos. Sin recomendaciones ni valores de referencia, pueden producirse desajustes alimentarios, que pueden asociarse con la aparición precoz de la mayor parte de las enfermedades crónicas o degenerativas en nuestra sociedad. Los objetivos de este trabajo son: clasificarlos hidratos de carbono presentes en los alimentos, establecer definiciones claras sobre todos los términos bioquímicos y comunes relacionados con los azúcares, explicar su valor nutricional y describir su metabolismo, así como las fuentes alimentarias que contienen tanto monocomo disacáridos y, finalmente, realizar un análisis DAFO(Debilidades, Amenazas, Fortalezas y Oportunidades)sobre la nomenclatura y las ingestas de azúcares (AU)

Carbohydrates are important and necessary components of human diet. Although they primarily play an energetic function, they also have structural and functional roles. According to the European Food Safety Authority, carbohydrate intake should range between 45 and 60percent of the energy in adults and children older than one year of age. An important part of carbohydrates available in foods are mono and disaccharides, commonly referred to as sugars. Dietary sources of sugars include fruits, fruit juices, vegetables, milk and milk products, and foods containing added sucrose and starch hydrolyzates. Despite their importance in daily life, there is currently no clear and adequate terminology on the various types of carbohydrates, particularly sugars. Nor are there available sugar intake recommendations or food composition tables. Without these recommendations or reference values, dietary unbalances might occur, which subsequently may end in the premature onset of most chronic or degenerative diseases of our society. The aims of the present work are: to classify dietary carbohydrates, to define the biochemical and common terms for sugars, to explain their nutritional value and their metabolism as well as their food sources and to carry out a SWOT (Strengths, Weaknesses, Opportunities, Threats) analysis about the nomenclature and dietary intakes of sugars (AU)

El azúcar en los distintos ciclos de la vida: desde la infancia hasta la vejez / Sugar role through the life cycle: from infancy to the elderly

Se revisa la importancia que tienen los diferentes tipos de azúcares consumidos a través de la dieta y en diferentes fuentes alimentarias en las etapas de la vida y situaciones fisiológicas especiales, y como el consumo moderado de azúcar es compatible con una dieta equilibrada y estilos de vida activos. Se describe también su función en el disfrute y placer de comer, como uno de los pilares básicos también de una alimentación equilibrada. Al mismo tiempo, se evalúan los riesgos del consumo muy insuficiente, principalmente en lo referido a un bajo aporte de glucosa en etapas críticas de la vida, así como las potenciales consecuencias negativas sobre el estado de salud en el ciclo vital cuando la ingesta es excesiva, y la asociación con factores de riesgo en enfermedades crónico-degenerativas, o con complicaciones en el embarazo. Finalmente, se aportan recomendaciones para un correcto consumo desde el punto de vista nutricional que constituyan, finalmente, herramientas válidas para una evaluación beneficio/riesgo a nivel individual o desde el punto de vista de la salud pública (AU)

The chapter reviews and updates the role of the different types of sugar along the life cycle, mainly during infancy and aging, but also for physiological situations such as pregnancy and breastfeeding. Moreover, several examples from infancy to the elderly illustrate that a moderate consumption may be considered as adequate within the context of a healthy diet and active life. In addition, the importance of sugar to provide palatability to the diet is also revised and attempted. The consequences of low sugar consumption (e.g. glucose) are also evaluated, but also the potential hazard effects of high and prolonged intakes at the different ages or pregnancy, aswell as its association with risk factors for chronic diseases. Finally, recommendations are given for adequate consumption to serve as tools for a benefit/risk evaluation at individual level and for public health strategies (AU)

Ayudas ergogénicas nutricionales para las personas que realizan ejercicio físico: Documento de Consenso de la Federación Española de Medicina del Deporte (FEMEDE) / No disponible

No disponible

Symbol nomenclature for representing glycan structures: Extension to cover different carbohydrate types.

This Viewpoint article addresses comments made on our original article describing a symbolic system for the depiction of N- and O-linked carbohydrate structures and proposes a method for extending the symbol set to include monosaccharides commonly found in carbohydrates present in bacteria and plants. As before, basic monosaccharides are shown by shape with one or more additions such as solid fill or additions of lines, crosses or dots to represent functional groups. The use of colour to differentiate constituent monosaccharides is avoided, thus enabling the system to be used in a variety of formats. Linkage and anomericity are shown by the angle and type of line connecting the symbols. In this extended version, new symbols are proposed for additional hexoses and it is proposed that pyranose and furanose forms of the monosaccharides could be shown by solid or broken outlines to the symbols. Conventions for depicting the presence of multiple functional groups such as deoxy-(NH(2))(2) are also discussed. It is hoped that these proposals will stimulate discussion so that a consensus can be reached as to how the glycobiology community can best convey complex information in a simple manner.

Los edulcorantes y su uso en Niños / Sweeteners and use in children

Los carbohidratos simples como el azúcar, se encuentran en una gran cantidad de alimentos como tortas,caramelos, helados, refrescos, gaseosas y bocadillos. Los edulcorantes son sustancias artificiales que se clasifican en nutritivos, y no nutritivos o no calóricos. Para que los edulcorantes fueran aprobados por la Food Drugs Administration (FDA), han pasado por una serie de pruebas farmacológicas y toxicológicas paradeterminar si su uso es seguro. Las dosis o cantidades seguras de consumo se denominan ingesta diariaaceptable o admisible que puede ser consumida por las personas en forma mantenida sin riesgo apreciable para la salud. Su uso de manera moderada, puede ser de gran utilidad en el manejo de una dieta balanceada o con disminución en las calorías totales, para conservar el peso adecuado o controlar la ganancia y mantenerniveles de glicemia lo más cercano a lo normal. Aún queda mucho por investigar en relación con los edulcorantes y los datos hasta el momento indican que son seguros.

Carbohydrates as the simple sugar found in a variety of foods such as cakes, candy, ice cream, soft drinks and snacks. Artificial sweeteners are substances that are classified as nutritive and non-nutritive or non-caloric. For sweeteners are approved by the Food Drugs Administration (FDA), have gone through a series of pharmacological and toxicological tests to determine if their use is safe. Safe doses or quantities ofconsumption are called acceptable daily intake or intake (ADI) that can be consumed by people in theform maintained without appreciable health risk. Its use in moderation, can be very useful in managing abalanced diet or decrease in total calories, to keep the weight or gain and maintain control of blood glucoselevels as close to normal. Much remains to be investigated in relation to sweeteners and the data so farindicate they are safe.

Stable isotope ratio measurements of royal jelly samples for controlling production procedures: impact of sugar feeding.

The carbon and nitrogen stable ratios of royal jelly (RJ) samples from various origins are determined using an elemental analyser linked online to an isotope ratio mass spectrometer to evaluate authenticity and adulteration. The (13)C/(12)C and (15)N/(14)N stable isotope ratios are measured in more than 500 RJs (domestic, imported and derived from feeding experiments) in order to obtain isotopic measurements that take into account seasonal, botanical and geographical effects. Authenticity intervals are established for traditional beekeeping practices, without feeding, in the range -22.48 to -27.90‰ for δ(13)C. For these samples, the δ(15)N values range from -1.58 to 7.98‰, depending on the plant sources of pollen and nectar. The δ(13)C values of the commercial samples vary from -18.54 to -26.58‰. High δ(13)C values are typical of sugar cane or corn syrups which have distinctive isotopic (13)C signatures because both plants use the C4 photosynthetic cycle, in contrast to most RJs which are derived from C3 plants. These differences in the (13)C-isotopic composition allow the detection of the addition of such sugars. RJs from traditional sources and from industrial production by sugar feeding are thus successfully distinguished.

Bacterial carbohydrate structure database 3: principles and realization.

Bacterial carbohydrate structure database (BCSDB) is an open-access project that collects primary publication data on carbohydrate structures originating from bacteria, their biological properties, bibliographic and taxonomic annotations, NMR spectra, etc. Almost complete coverage and outstanding data consistency are achieved. BCSDB version 3 and the principles lying behind it, including glycan description language, are reported.