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PloS One 2022O-GlcNAcylation is the only sugar modification for proteins present in the cytoplasm and nucleus and is thought to be involved in the regulation of protein function and...
O-GlcNAcylation is the only sugar modification for proteins present in the cytoplasm and nucleus and is thought to be involved in the regulation of protein function and localization. Currently, several methods are known for detecting O-GlcNAcylated proteins using monoclonal antibodies or wheat germ agglutinin, but these methods have some limitations in their sensitivity and quantitative comparison. We developed a new disaccharide-tag method to overcome these problems. This is a method in which a soluble GalNAc transferase is expressed intracellularly, extended to a disaccharide of GalNAc-GlcNAc, and detected using a Wisteria japonica agglutinin specific to this disaccharide. We verified the method using human c-Rel protein and also highly sensitively compared the difference in O-GlcNAc modification of intracellular proteins associated with differentiation from embryonic stem cell (ESC) to epiblast-like cells (EpiLC). As one example of such a modification, a novel O-GlcNAc modification was found in the transcription factor Sox2 at residue Ser263, and the modification site could be identified by nano liquid chromatography-mass spectrometry.
Topics: Acetylglucosamine; Animals; Disaccharides; Glycosylation; Humans; Mammals; Mass Spectrometry; N-Acetylglucosaminyltransferases; Protein Processing, Post-Translational; Proteins
PubMed: 35604954
DOI: 10.1371/journal.pone.0267804 -
Biomacromolecules Jun 2021Antifreeze glycoproteins (AFGPs) are able to bind to ice, halt its growth, and are the most potent inhibitors of ice recrystallization known. The structural basis for...
Antifreeze glycoproteins (AFGPs) are able to bind to ice, halt its growth, and are the most potent inhibitors of ice recrystallization known. The structural basis for AFGP's unique properties remains largely elusive. Here we determined the antifreeze activities of AFGP variants that we constructed by chemically modifying the hydroxyl groups of the disaccharide of natural AFGPs. Using nuclear magnetic resonance, two-dimensional infrared spectroscopy, and circular dichroism, the expected modifications were confirmed as well as their effect on AFGPs solution structure. We find that the presence of all the hydroxyls on the disaccharides is a requirement for the native AFGP hysteresis as well as the maximal inhibition of ice recrystallization. The saccharide hydroxyls are apparently as important as the acetyl group on the galactosamine, the α-linkage between the disaccharide and threonine, and the methyl groups on the threonine and alanine. We conclude that the use of hydrogen-bonding through the hydroxyl groups of the disaccharide and hydrophobic interactions through the polypeptide backbone are equally important in promoting the antifreeze activities observed in the native AFGPs. These important criteria should be considered when designing synthetic mimics.
Topics: Antifreeze Proteins; Disaccharides; Glycoproteins; Hydrogen Bonding; Ice; Magnetic Resonance Spectroscopy
PubMed: 33957041
DOI: 10.1021/acs.biomac.1c00313 -
Applied Microbiology and Biotechnology May 2021In a competitive microbial environment, nutrient acquisition is a major contributor to the survival of any individual bacterial species, and the ability to access... (Review)
Review
In a competitive microbial environment, nutrient acquisition is a major contributor to the survival of any individual bacterial species, and the ability to access uncommon energy sources can provide a fitness advantage. One set of soluble carbohydrates that have attracted increased attention for use in biotechnology and biomedicine is the α-diglucosides. Maltose is the most well-studied member of this class; however, the remaining four less common α-diglucosides (trehalose, kojibiose, nigerose, and isomaltose) are increasingly used in processed food and fermented beverages. The consumption of trehalose has recently been shown to be a contributing factor in gut microbiome disease as certain pathogens are using α-diglucosides to outcompete native gut flora. Kojibiose and nigerose have also been examined as potential prebiotics and alternative sweeteners for a variety of foods. Compared to the study of maltose metabolism, our understanding of the synthesis and degradation of uncommon α-diglucosides is lacking, and several fundamental questions remain unanswered, particularly with regard to the regulation of bacterial metabolism for α-diglucosides. Therefore, this minireview attempts to provide a focused analysis of uncommon α-diglucoside metabolism in bacteria and suggests some future directions for this research area that could potentially accelerate biotechnology and biomedicine developments. KEY POINTS: • α-diglucosides are increasingly important but understudied bacterial metabolites. • Kinetically superior α-diglucoside enzymes require few amino acid substitutions. • In vivo studies are required to realize the biotechnology potential of α-diglucosides.
Topics: Bacteria; Biotechnology; Isomaltose; Maltose; Trehalose
PubMed: 33961116
DOI: 10.1007/s00253-021-11322-x -
Journal of Animal Science Feb 2022A completely randomized design employing a 2 × 2 factorial experiment was designed in this study to evaluate the effects of in ovo injection of disaccharide (DS)...
A completely randomized design employing a 2 × 2 factorial experiment was designed in this study to evaluate the effects of in ovo injection of disaccharide (DS) and/or methionine (Met) on hatchability, growth performance, blood hematology, and serum antioxidant parameters in geese. A total of 600 fertilized geese's eggs containing live embryo were randomly assigned into 4 groups with 6 replicates and 25 eggs per replicate. Factors in four groups comprised noninjection, DS injection (25 g/L maltose + 25 g/L sucrose + 7.5 g/L NaCl), Met injection (5 g/L Met + 7.5 g/L NaCl), or DS plus Met injection (25 g/L maltose + 25 g/L sucrose + 5 g/L Met + 7.5 g/L NaCl), respectively. We found that the administration of DS in embryo increased hatching time, yolk sac-free carcass weight, yolk sac-free carcass indexes and decreased assisted hatching ratio, yolk sac weight, yolk sac indexes, but did not affect hatchability and mortality. Moreover, higher body weight and serum glucose concentrations in DS injection group compared with noninjection group were observed on day of hatching. The body weight and average daily gain (ADG) of geese in DS injection group were higher than noninjection group after incubation. In ovo injection of Met increased hatching time and yolk sac-free carcass indexes, but decreased yolk sac indexes. In addition, the strategy of in ovo feeding of Met led to higher body weight, ADG, serum uric acid, glutathione (GSH), and glutathione peroxidase concentrations, as well as lower GSSG/GSH ratio, serum glutathione disulfide (GSSG), and malondialdehyde (MDA) concentrations than the noninjection group on day of hatching. The post-hatching body weight, ADG, serum total protein, albumin, and uric acid concentrations increased, whereas post-hatching serum GSSG and MDA concentrations and GSSG/GSH ratio decreased when injected with Met. In addition, synergistic effects of in ovo injection of DS plus Met on hatching time as well as post-hatching body weight and ADG were observed. Therefore, in ovo injection of DS plus Met was demonstrated to be a way to improve the development of geese during early incubation stages.
Topics: Animals; Antioxidants; Chickens; Disaccharides; Geese; Hematology; Methionine; Ovum; Uric Acid
PubMed: 35094079
DOI: 10.1093/jas/skac014 -
Frontiers in Endocrinology 2022The non-absorbable disaccharide lactulose is mostly used in the treatment of various gastrointestinal disorders such as chronic constipation and hepatic encephalopathy.... (Review)
Review
The non-absorbable disaccharide lactulose is mostly used in the treatment of various gastrointestinal disorders such as chronic constipation and hepatic encephalopathy. The mechanism of action of lactulose remains unclear, but it elicits more than osmotic laxative effects. As a prebiotic, lactulose may act as a bifidogenic factor with positive effects in preventing and controlling diabetes. In this review, we summarized the current evidence for the effect of lactulose on gut metabolism and type 2 diabetes (T2D) prevention. Similar to acarbose, lactulose can also increase the abundance of the short-chain fatty acid (SCFA)-producing bacteria and as well as suppress the potentially pathogenic bacteria . These bacterial activities have anti-inflammatory effects, nourishing the gut epithelial cells and providing a protective barrier from microorganism infection. Activation of peptide tyrosine tyrosine (PYY) and glucagon-like peptide 1 (GLP1) can influence secondary bile acids and reduce lipopolysaccharide (LPS) endotoxins. A low dose of lactulose with food delayed gastric emptying and increased the whole gut transit times, attenuating the hyperglycemic response without adverse gastrointestinal events. These findings suggest that lactulose may have a role as a pharmacotherapeutic agent in the management and prevention of type 2 diabetes actions on the gut microbiota.
Topics: Acarbose; Anti-Inflammatory Agents; Bacteria; Bile Acids and Salts; Diabetes Mellitus, Type 2; Fatty Acids, Volatile; Glucagon-Like Peptide 1; Humans; Lactulose; Laxatives; Lipopolysaccharides; Peptides; Tyrosine
PubMed: 36187096
DOI: 10.3389/fendo.2022.956203 -
Angewandte Chemie (International Ed. in... Mar 2020C-Glycosides are both a common motif in many bioactive natural products and important glycoside mimetics. We demonstrate that activating a hemiacetal with a sulfonyl...
C-Glycosides are both a common motif in many bioactive natural products and important glycoside mimetics. We demonstrate that activating a hemiacetal with a sulfonyl chloride, followed by treating the resultant glycosyl sulfonate with an enolate results in the stereospecific construction of β-linked C-glycosides. This reaction tolerates a range of acceptors and donors, including disaccharides. The resulting products can be readily derivatized into C-glycoside analogues of β-glycoconjugates, including C-disaccharide mimetics.
Topics: Alkylation; Disaccharides; Glycoconjugates; Glycosides; Glycosylation; Molecular Structure; Stereoisomerism; Sulfinic Acids
PubMed: 31880395
DOI: 10.1002/anie.201914221 -
BioEssays : News and Reviews in... Nov 2022Neurological diseases (NDs), featured by progressive dysfunctions of the nervous system, have become a growing burden for the aging populations. N-Deacetylase and...
Neurological diseases (NDs), featured by progressive dysfunctions of the nervous system, have become a growing burden for the aging populations. N-Deacetylase and N-sulfotransferase 3 (NDST3) is known to catalyze deacetylation and N-sulfation on disaccharide substrates. Recently, NDST3 is identified as a novel deacetylase for tubulin, and its newly recognized role in modulating microtubule acetylation and lysosomal acidification provides fresh insights into ND therapeutic approaches using NDST3 as a target. Microtubule acetylation and lysosomal acidification have been reported to be critical for activities in neurons, implying that the regulators of these two biological processes, such as the previously known microtubule deacetylases, histone deacetylase 6 (HDAC6) and sirtuin 2 (SIRT2), could play important roles in various NDs. Aberrant NDST3 expression or tubulin acetylation has been observed in an increasing number of NDs, including amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD), schizophrenia and bipolar disorder, Alzheimer's disease (AD), and Parkinson's disease (PD), suggesting that NDST3 is a key player in the pathogenesis of NDs and may serve as a target for development of new treatment of NDs.
Topics: Humans; Amyotrophic Lateral Sclerosis; Disaccharides; Frontotemporal Dementia; Hydrogen-Ion Concentration; Lysosomes; Microtubules; Sirtuin 2; Sulfotransferases; Tubulin
PubMed: 36135988
DOI: 10.1002/bies.202200110 -
Bioscience Reports Sep 2023Iron deficiency anemia (IDA) is a leading global health concern affecting approximately 30% of the population. Treatment for IDA consists of replenishment of iron...
Iron deficiency anemia (IDA) is a leading global health concern affecting approximately 30% of the population. Treatment for IDA consists of replenishment of iron stores, either by oral or intravenous (IV) supplementation. There is a complex bidirectional interplay between the gut microbiota, the host's iron status, and dietary iron availability. Dietary iron deficiency and supplementation can influence the gut microbiome; however, the effect of IV iron on the gut microbiome is unknown. We studied how commonly used IV iron preparations, ferric carboxymaltose (FCM) and ferric derisomaltose (FDI), affected the gut microbiome in female iron-deficient anemic mice. At the phylum level, vehicle-treated mice showed an expansion in Verrucomicrobia, mostly because of the increased abundance of Akkermansia muciniphila, along with contraction in Firmicutes, resulting in a lower Firmicutes/Bacteroidetes ratio (indicator of dysbiosis). Treatment with either FCM or FDI restored the microbiome such that Firmicutes and Bacteroidetes were the dominant phyla. Interestingly, the phyla Proteobacteria and several members of Bacteroidetes (e.g., Alistipes) were expanded in mice treated with FCM compared with those treated with FDI. In contrast, several Clostridia class members were expanded in mice treated with FDI compared with FCM (e.g., Dorea spp., Eubacterium). Our data demonstrate that IV iron increases gut microbiome diversity independently of the iron preparation used; however, differences exist between FCM and FDI treatments. In conclusion, replenishing iron stores with IV iron preparations in clinical conditions, such as inflammatory bowel disease or chronic kidney disease, could affect gut microbiome composition and consequently contribute to an altered disease outcome.
Topics: Female; Animals; Mice; Iron; Gastrointestinal Microbiome; Disaccharides; Iron, Dietary; Bacteroidetes; Firmicutes
PubMed: 37671923
DOI: 10.1042/BSR20231217 -
Food Research International (Ottawa,... Feb 2023The effect of monosaccharides (glucose, fructose and galactose) and disaccharides (sucrose and lactose) at 10, 20 and 30 % w/v on the in-vitro aroma partitioning of C -...
The effect of monosaccharides (glucose, fructose and galactose) and disaccharides (sucrose and lactose) at 10, 20 and 30 % w/v on the in-vitro aroma partitioning of C - C aldehydes and ethyl esters, as well as limonene (concentration of aroma compounds at 1 μg mL), was studied using atmospheric pressure chemical ionisation-mass spectrometry. An increase in sugar concentration from 0 to 30 % w/v resulted in a significant increase in partitioning under static headspace conditions for the majority of the compounds (p < 0.05), an effect generally not observed when 10 % w/v sucrose was substituted with low-calorie sweeteners (p > 0.05). The complexity of the system was increased to model a soft drink design - comprising water, sucrose (10, 20 and 30 % w/v), acid (0.15 % w/v), carbonation (∼7.2 g/L CO) and aroma compounds representative of an apple style flavouring, namely ethyl butanoate and hexanal (10 μg mL each). Although the addition of sucrose had no significant in-vivo effect, carbonation significantly decreased breath-by-breath (in-vivo) aroma delivery (p < 0.05). To understand the physical mechanisms behind aroma release from the beverage matrix, the effect of sucrose on the kinetics of the matrix components was explored. An increase in sucrose concentration from 0 to 30 % w/v resulted in a significant decrease in water activity (p < 0.05), which accounted for the significantly slower rate of self-diffusion of aroma compounds (p < 0.05), measured using diffusion-ordered spectroscopy-nuclear magnetic resonance spectroscopy. No significant effect of sucrose on carbon dioxide volume flux was found (p > 0.05).
Topics: Odorants; Sweetening Agents; Sucrose; Magnetic Resonance Spectroscopy; Beverages; Water
PubMed: 36737960
DOI: 10.1016/j.foodres.2022.112373 -
MSphere Apr 2022Yeast species in the and genera (W/S clade) thrive in the sugar-rich floral niche. We have previously shown that species belonging to this clade harbor an unparalleled...
Yeast species in the and genera (W/S clade) thrive in the sugar-rich floral niche. We have previously shown that species belonging to this clade harbor an unparalleled number of genes of bacterial origin, among which is the gene, encoding a sucrose-hydrolyzing enzyme. In this study, we used complementary and experimental approaches to examine sucrose utilization in a broader cohort of species representing extant diversity in the W/S clade. Distinct strategies and modes of sucrose assimilation were unveiled, involving either extracellular sucrose hydrolysis through secreted bacterial Suc2 or intracellular assimilation using broad-substrate-range α-glucoside/H symporters and α-glucosidases. The intracellular pathway is encoded in two types of gene clusters reminiscent of the clusters in Saccharomyces cerevisiae, where they are involved in maltose utilization. The genes composing each of the two types of clusters found in the W/S clade have disparate evolutionary histories, suggesting that they formed . Both transporters and glucosidases were shown to be functional and additionally involved in the metabolization of other disaccharides, such as maltose and melezitose. In one species lacking the α-glucoside transporter, maltose assimilation is accomplished extracellularly, an attribute which has been rarely observed in fungi. Sucrose assimilation in generally escaped both glucose repression and the need for an activator and is thus essentially constitutive, which is consistent with the abundance of both glucose and sucrose in the floral niche. The notable plasticity associated with disaccharide utilization in the W/S clade is discussed in the context of ecological implications and energy metabolism. Microbes usually have flexible metabolic capabilities and are able to use different compounds to meet their needs. The yeasts belonging to the and genera (forming the so-called W/S clade) are usually found in flowers or insects that visit flowers and are known for having acquired many genes from bacteria by a process called horizontal gene transfer. One such gene, dubbed , is used to assimilate sucrose, which is one of the most abundant sugars in floral nectar. Here, we show that different lineages within the W/S clade used different solutions for sucrose utilization that dispensed and differed in their energy requirements, in their capacity to scavenge small amounts of sucrose from the environment, and in the potential for sharing this resource with other microbial species. We posit that this plasticity is possibly dictated by adaptation to the specific requirements of each species.
Topics: Glucose; Glucosides; Humans; Maltose; Saccharomyces cerevisiae; Saccharomycetales; Sucrose
PubMed: 35354279
DOI: 10.1128/msphere.00035-22