Mannose Oligosaccharides Research Articles

Treatment with mannose oligosaccharides reverses the intestinal injury in the acetylsalicylic acid-treated rat model

Mannose oligosaccharide (MOS) has been shown to promote animal growth, maintain intestinal health, and activate the intestinal immune system. However, the question of whether MOS can stimulate the immune system and alleviate acetylsalicylic acid (ASA)-induced gut damage remains unresolved. The purpose of this study was to investigate the impact of MOS pretreatment on the immunological and anti-inflammatory capabilities of rats with ASA-induced intestinal injury. Thirty-six male Sprague-Dawley rats were divided into 6 groups and were fed with 0 (negative control), 100, 300, 600, and 800 mg/kg·Body weight (BW) of MOS for 3 weeks. From day 8, rats were fed with 200 mg/kg BW of ASA for 14 days to induce intestinal injury. The growth performance, viscera index, serum and intestinal immunity, intestinal inflammation and morphology of ASA-induced intestinal injury rats with or without MOS administration were investigated. In MOS deficient rats, oral treatment of ASA causes severe intestine damage and immunological dysfunction. In a rat model, 600 mg/kg BW MOS can lower the expression of inflammatory markers and effectively increase liver index, serum interleukin-2 (IL-2), lysozyme contents, intestinal secretory immunoglobulin A (sIgA) and mucus volume, intestinal villus height, crypt depth and villus height/crypt depth in comparison to the ASA group. These results imply that providing rats with MOS at the appropriate dosage can significantly improve their immune system and successfully shield the intestines from ASA damage. MOS is therefore expected to be a promising gut immunopotentiator for enhancing intestinal health in animals.

Effects of a novel synbiotics-enzyme complex as a replacement for antibiotics on growth performance, slaughter and meat characteristics, immune organ index, and intestinal morphology of broilers.

Antibiotic use in broilers is being discouraged globally due to the challenges it poses. This study was conducted to assess the effects of supplementing broilers with a Symbiotic-Enzyme complex (SEC) containing prebiotics (mannose oligosaccharides), probiotics (Clostridium butyricum and Bacillus subtilis), and enzymes (glucose oxidase, and α-galactosidase) as an alternative to antibiotics on growth performance, carcass and meat quality traits, mortality, linear body measurements, intestinal morphology and immune organ indexes. A total of 864 mixed-sex 1-day-old arbor acres (AA+) broilers were allocated to 8 experimental groups replicated 9 times with 12 chickens per replicate. These included 6 treatment groups with SEC inclusion levels of 0.025, 0.04, 0.05, 0.06, 0.08, and 0.10%, respectively, and two control groups: a negative control group containing a basal diet only and the positive control group (Antibiotics group) containing a basal diet and antibiotic oxytetracycline added at 0.2%. Growth performance was measured on day 21 and 42, and the mortality, carcass, meat quality traits, linear body measurements, intestinal morphology, and organ size indexes were measured on day 42. The results indicated that supplementing broilers with 0.1% SEC resulted in insignificant (P > 0.05) increases in average daily feed intake (ADFI), significant (P < 0.05) increases in the average daily gains (ADG), and significant (P < 0.05) reduction in a feed-to-gain ratio (F/G) in all the phases compared to the control and antibiotics groups. Supplementation of broilers with 0.1% SEC inclusion levels also significantly (P < 0.05) increased the body slope length, chest width, chest depth, keel length, and shank circumference. Furthermore, broilers on diets containing 0.1% SEC inclusion level also had significantly (P < 0.05) higher dressed, semi-evisceration, evisceration, and breast muscle percentages. Including SEC at 0.1% also significantly (P < 0.05) increased villus height and villus-to-crypt ratio (V/C) but reduced crypt depth in the duodenum, jejunum, and ileum compared to the control groups. SEC inclusion at 0.1% significantly (P < 0.05) increased the spleen, bursal, and thymus indexes, respectively. Supplementation of broilers with 0.1% SEC can be used as an antibiotic alternative because it increases the F/G, improves the carcass and meat quality, increases the body conformation, improves the small intestines' functions, and immune organ size.

Mannose oligosaccharide recognition of CGL1, a mannose-specific lectin containing DM9 motifs from Crassostrea gigas, revealed by X-ray crystallographic analysis.

CGL1 is a mannose-specific lectin isolated from the Pacific oyster Crassostrea gigas, and it belongs to the DM9 domain protein family. Each subunit of the CGL1 dimer consists of a tandem repeat of DM9 motifs, which were originally found in the Drosophila melanogaster genome. The CGL1 protomer contains two carbohydrate-binding sites: a high-affinity site A and a low-affinity site B. An assay using dendrimers containing oligomannose from yeast (Saccharomyces cerevisiae) revealed that CGL1 exhibited significantly higher affinity for mannotetraose (Man4) compared to mannobiose (Man2) and mannotriose (Man3). To investigate its oligomannose-recognition mechanism, X-ray crystallographic analyses of CGL1/oligomannose complexes were performed. In the CGL1/Man2 and CGL1/Man3 complexes, Manα1-2Man and Manα1-2Manα1-2Man, respectively, were primarily bound to site A, interacting with the non-reducing mannose residue. On the other hand, in the CGL1/Man4 crystal, Man4 (Manα1-2Manα1-2Manα1-6Man) was bound at both site A and site B at the non-reducing and reducing ends, thus linking adjacent CGL1 molecules with crystallographic symmetry. These findings suggest that CGL1 can recognize both the non-reducing and reducing mannose residues of mannose oligosaccharides at its two distinct carbohydrate-binding sites. This enables efficient complex formation, making CGL1 a pattern-recognition molecule capable of recognizing diverse structures of mannose-containing carbohydrate chains.

Effects of nonantibiotic growth promoter combinations on growth performance, nutrient utilization, digestive enzymes, intestinal morphology, and cecal microflora of broilers.

Given the ban on antibiotic growth promoters, the effects of nonantibiotic alternative growth promoter combinations (NAGPCs) on the growth performance, nutrient utilization, digestive enzyme activity, intestinal morphology, and cecal microflora of broilers were evaluated. All birds were fed pellets of two basal diets-starter (0-21 d) and grower (22-42 d)-with either enramycin (ENR) or NAGPC supplemented. 1) control + ENR; 2) control diet (CON, basal diet); 3) control + mannose oligosaccharide (MOS) + mannanase (MAN) + sodium butyrate (SB) (MMS); 4) control + MOS + MAN + Bacillus subtilis (BS) (MMB); 5) control + MOS + fruit oligosaccharide (FOS) + SB (MFS); 6) control + FOS + BS (MFB); 7) control + MOS + FOS + MAN (MFM); 8) control + MOS + BS + phytase (PT) (MBP). ENR, MOS, FOS, SB, MAN, PT, and BS were added at 100, 2,000, 9,000, 1,500, 300, 37, and 500 mg/kg, respectively. The experiment used a completely random block design with six replicates per group: 2400 Ross 308 broilers in the starter phase and 768 in the grower phase. All NAGPCs significantly improved body weight gain (P < 0.01), utilization of dry matter, organic matter, and crude protein (P < 0.05), villus height and villus height/crypt depth in the jejunum and ileum (P < 0.01), and decreased the feed conversion ratio (P < 0.01) at d 21 and 42. MMS, MMB, MFB, and MFM duodenum trypsin, lipase, and amylase activities increased significantly (P < 0.05) at d 21 and 42. On d 21 and 42, MMS, MMB, and MBP increased the abundance of Firmicutes and Bacteroides whereas MMB, MFB, and MBP decreased the abundance of Proteobacteria, compared to ENR and CON. Overall, the NAGPCs were found to have some beneficial effects and may be used as effective antibiotic replacements in broilers.

Dietary mannan oligosaccharides strengthens intestinal immune barrier function via multipath cooperation during Aeromonas Hydrophila infection in grass carp (Ctenopharyngodon Idella).

In recent years, mannose oligosaccharide (MOS) as a functional additive is widely used in aquaculture, to enhance fish immunity. An evaluation of the effect of dietary MOS supplementation on the immune barrier function and related signaling molecules mechanism of grass carp (Ctenopharyngodon idella) was undertaken in the present study. Six diets with graded amounts of MOS supplementation (0, 200, 400, 600, 800, and 1000 mg/kg) were fed to 540 grass carp over 60 days. To examine the immune response and potential mechanisms of MOS supplementation on the intestine, a challenge test was conducted using injections of Aeromonas hydrophila for 14 days. Results of the study on the optimal supplementation with MOS were found as follows (1) MOS enhances immunity partly related to increasing antibacterial substances content and antimicrobial peptides expression; (2) MOS attenuates inflammatory response partly related to regulating the dynamic balance of intestinal inflammatory cytokines; (3) MOS regulates immune barrier function may partly be related to modulating TLRs/MyD88/NFκB and TOR/S6K1/4EBP signalling pathways. Finally, the current study concluded that MOS supplementation could improve fish intestinal immune barrier function under Aeromonas hydrophila infected conditions.

Effect of Dietary Supplementation with Mannose Oligosaccharides on the Body Condition, Lactation Performance and Their Offspring of Heat-Stressed Sows.

Simple SummarySummer heat stress (HS) seriously affects the reproductive and lactation performance of sows and the long-term development of their offspring. Mannose oligosaccharide (MOS) is widely used as an ingredient in animal feed as it can limit the colonization of enteric pathogens. However, most animal experiments that use MOS to alleviate the effects of HS are often performed with broilers. The effect of MOS in sows affected by HS is unclear. Based on our results, dietary supplementation with MOS tended to alleviate HS in sows compared with the control. However, no significant interactive effects were found.The aim of this study was to determine the effects of dietary supplementation with mannose oligosaccharide (MOS) on the condition of the body and the reproductive and lactation performances of sows. Eighty pregnant sows were randomly assigned to four groups with a 2 × 2 factorial design: with or without MOS (1 g/kg) and with or without heat stress (HS) challenge. The temperature in the HS groups (HS and HM group) was controlled at 31.56 ± 1.22 °C, while the temperature in the active cooling (AC) groups (AC and AM group) was controlled at 23.49 ± 0.72 °C. The weight loss of sows in the AC group was significantly lower than that of sows in the HS group (p < 0.01). The weight and backfat thickness loss of sows supplemented with MOS displayed a downward trend. The average birth weight of the litter significantly increased in the HM group (basic diet + MOS) compared with the HS group (p < 0.05). The milk protein of sows significantly decreased under the HS condition at 2 and 12 h after delivery (p < 0.05). However, the milk immunoglobin G (IgG) of sows in the HS group increased significantly compared with that of sows in the HM group (p < 0.05) at 12 and 24 h after delivery. The levels of serum urea nitrogen (UREA) and glucose (GLU) decreased significantly under the HS condition (p < 0.05), while the level of interleukin-6 (IL-6) increased significantly under the HS condition (p < 0.05). Dietary supplementation with MOS also significantly reduced TNF-α under the AC conditions (p < 0.05). In conclusion, HS significantly affected the body condition, lactation performances and their offspring of sows. However, dietary supplementation with 1 g/kg MOS did not result in statistically significant changes.

Deploying an In Vitro Gut Model to Assay the Impact of the Mannan-Oligosaccharide Prebiotic Bio-Mos on the Atlantic Salmon (Salmo salar) Gut Microbiome.

ABSTRACTAlpha mannose-oligosaccharide (MOS) prebiotics are widely deployed in animal agriculture as immunomodulators as well as to enhance growth and gut health. Their mode of action is thought to be mediated through their impact on host microbial communities and their associated metabolism. Bio-Mos is a commercially available prebiotic currently used in the agri-feed industry, but studies show contrasting results of its effect on fish performance and feed efficiency. Thus, detailed studies are needed to investigate the effect of MOS supplements on the fish microbiome to enhance our understanding of the link between MOS and gut health. To assess Bio-Mos for potential use as a prebiotic growth promoter in salmonid aquaculture, we have modified an established Atlantic salmon in vitro gut model, SalmoSim, to evaluate its impact on the host microbial communities. The microbial communities obtained from ceca compartments from four adult farmed salmon were inoculated in biological triplicate reactors in SalmoSim. Prebiotic treatment was supplemented for 20 days, followed by a 6-day washout period. Inclusion of Bio-Mos in the media resulted in a significant increase in formate (P = 0.001), propionate (P = 0.037) and 3-methyl butanoic acid (P = 0.024) levels, correlated with increased abundances of several, principally, anaerobic microbial genera (Fusobacterium, Agarivorans, Pseudoalteromonas). DNA metabarcoding with the 16S rDNA marker confirmed a significant shift in microbial community composition in response to Bio-Mos supplementation with observed increase in lactic acid producing Carnobacterium. In conjunction with previous in vivo studies linking enhanced volatile fatty acid production alongside MOS supplementation to host growth and performance, our data suggest that Bio-Mos may be of value in salmonid production. Furthermore, our data highlights the potential role of in vitro gut models to complementin vivo trials of microbiome modulators.IMPORTANCE In this paper we report the results of the impact of a prebiotic (alpha-MOS supplementation) on microbial communities, using an in vitro simulator of the gut microbial environment of the Atlantic salmon. Our data suggest that Bio-Mos may be of value in salmonid production as it enhances volatile fatty acid production by the microbiota from salmon pyloric ceca and correlates with a significant shift in microbial community composition with observed increase in lactic acid producing Carnobacterium. In conjunction with previous in vivo studies linking enhanced volatile fatty acid production alongside MOS supplementation to host growth and performance, our data suggest that Bio-Mos may be of value in salmonid production. Furthermore, our data highlights the potential role of in vitro gut models to augment in vivo trials of microbiome modulators.

Investigating the effects of peptide-based, MOS and protease feed additives on the growth performance and fecal microbial composition of weaned pigs

BackgroundDigestive disorders in weaning pigs remain a major challenge for swine producers. Different types of commercial feed additives have been developed to promote gut health and development in young pigs, but their effects on resident gut microbial communities remain largely unexplored. The aim of this study was to investigate the impact of a peptide-based product (Peptiva) in combination with mannose oligosaccharides (MOS) and an exogenous protease on the performance and fecal microbiome of nursery pigs.MethodsA total of 1097 weaned pigs were divided into 44 pens (24–26 pigs/pen) with each pen randomly assigned to one of four experimental diets as part of Phase II and Phase III of a standard nursery phase feeding program. Fecal samples collected from representative control and treatment pigs were used to investigate bacterial composition profiles by high throughput sequencing of PCR-generated amplicons targeting the V1-V3 region of the 16S rRNA gene.ResultsHigher gain:feed was observed for pigs fed Peptiva and MOS compared to Controls during the period when experimental diets were fed, but the benefits of supplementation were not maintained after pigs were transitioned to a non-supplemented diet. Three candidate bacterial species, identified as Operational Taxonomic Units (OTUs), were found to have significantly different abundances between control samples and treatment samples during the same phase. In Phase III samples, SD_Ssd-00039, predicted to be a strain of Streptococcus alactolyticus based on nucleotide sequence identity, was the most highly represented of these OTUs with an average abundance in pigs fed Peptiva, MOS and protease that was 3.9 times higher than in Controls. The report also presents evidence of microbial succession that occurred during the trial, with 16 of the 32 most abundant OTUs found to vary between Phase II and Phase III samples for the same dietary treatment.ConclusionsDietary supplementation with a combination of a peptide-based product, MOS, and protease increased the growth performance of weaned pigs compared to control animals during the nursery phase, but these benefits were no longer observed within 2 weeks after all animals were transitioned to a non-supplemented diet. Supplementation with these feed additives was found to modulate the composition of the swine gut microbiome during this period.

Upgrading the Nutritional Value of PKC Using a Bacillus subtilis Derived Monocomponent β-Mannanase.

Palm kernel cake (PKC) is an abundant side stream that can only be added to non-ruminant feed in small concentrations due to its content of antinutritional factors, mainly galactomannan, which cannot be digested by non-ruminants. β-mannanases can be added to partially hydrolyze galactomannan to form mannose oligosaccharides, which are known to be prebiotic. We here investigate the action of a β-mannanase from B. subtilis on PKC by colorimetry, NMR and fluorescence microscopy. The amount of mannan oligosaccharides in solution was significantly increased by the β-mannanase and their degree of polymerization (DP) was significantly reduced. There was a dose-response behavior in that larger β-mannanase concentrations increased the amount of soluble mannose oligosaccharides while reducing their average DP. Using confocal immunofluorescence microscopy, solubilization of galactomannan in PKC was clearly visualized. Images show a clear disruption of the cellulose and galactomannan structures of the PKC cell walls. We thus show in this study that using commercial dosages of β-mannanase on PKC can lead to formation of prebiotic compounds. Thus, this study suggests that utilization of PKC in poultry feed formulation might be increased by addition of a β-mannanase and would improve the return on investment.

Combining oligomer build-up with alanine scanning to determine the flocculation protein mutants for enhancing oligosaccharide binding

ABSTRACT Oligosaccharide binding on protein plays an important role in cellular signalling and recognition. Protein mutation can alter the binding strength and thus regulate the relevant processes. In this work, we investigated the interactions between the mannose oligosaccharides and the wild type and mutant forms of yeast flocculation protein Flo1p from Saccharomyces cerevisiae. The binding strength of an oligosaccharide chain was estimated using molecular dynamics simulation and MM/GBSA binding free energy calculation, via the build-up of oligosaccharide from monosaccharide to disaccharide, trisaccharide and tetrasaccharide. At the protein-disaccharide stage, several mutation positions were identified via alanine scanning mutagenesis, leading to nine mutants, from which two mutants Q117N and Q117R were selected with significantly stronger binding free energies relative to the wild type protein. Furthermore, the two mutants also show enhanced binding at the trisaccharide and tetrasaccharide stages. Conformational stability and shape change were analysed to help understand the protein-oligosaccharide interactions. This study also presented a computational example to estimate the oligosaccharide binding strength using its terminal sugar residues.

Disruption of protein rhamnosylation affects the Sporothrix schenckii-host interaction

Sporotrichosis is a fungal disease caused by the members of the Sporothrix pathogenic clade, and one of the etiological agents is Sporothrix schenckii. The cell wall of this organism has been previously analyzed and thus far is known to contain an inner layer composed of chitin and β -glucans, and an outer layer of glycoproteins, which are decorated with mannose and rhamnose-containing oligosaccharides. The L-rhamnose biosynthesis pathway is common in bacteria but rare in members of the Fungi kingdom. Therefore, in this study, we aimed to disrupt this metabolic route to assess the contribution of rhamnose during the S.schenckii-host interaction. We identified and silenced in S. schenckii a functional ortholog of the bacterial rmlD gene, which encodes for an essential reductase for the synthesis of nucleotide-activated L-rhamnose. RmlD silencing did not affect fungal growth or morphology but decreased cell wall rhamnose content. Compensatory, the β-1,3-glucan levels increased and were more exposed at the cell surface. Moreover, when incubated with human peripheral blood mononuclear cells, the RmlD silenced mutants differentially stimulated cytokine production when compared with the wild-type strain, reducing TNFα and IL-6 levels and increasing IL-1 β and IL-10 production. Upon incubation with human monocyte-derived macrophages, the silenced strains were more efficiently phagocytosed than the wild-type strain. In both cases, our data suggest that rhamnose-based oligosaccharides are ligands that interact with TLR4. Finally, our findings showed that cell wall rhamnose is required for the S. schenckii virulence in the G. mellonella model of infection.

Antiviral Cyanometabolites-A Review.

Global processes, such as climate change, frequent and distant travelling and population growth, increase the risk of viral infection spread. Unfortunately, the number of effective and accessible medicines for the prevention and treatment of these infections is limited. Therefore, in recent years, efforts have been intensified to develop new antiviral medicines or vaccines. In this review article, the structure and activity of the most promising antiviral cyanobacterial products are presented. The antiviral cyanometabolites are mainly active against the human immunodeficiency virus (HIV) and other enveloped viruses such as herpes simplex virus (HSV), Ebola or the influenza viruses. The majority of the metabolites are classified as lectins, monomeric or dimeric proteins with unique amino acid sequences. They all show activity at the nanomolar range but differ in carbohydrate specificity and recognize a different epitope on high mannose oligosaccharides. The cyanobacterial lectins include cyanovirin-N (CV-N), scytovirin (SVN), microvirin (MVN), Microcystis viridis lectin (MVL), and Oscillatoria agardhii agglutinin (OAA). Cyanobacterial polysaccharides, peptides, and other metabolites also have potential to be used as antiviral drugs. The sulfated polysaccharide, calcium spirulan (CA-SP), inhibited infection by enveloped viruses, stimulated the immune system’s response, and showed antitumor activity. Microginins, the linear peptides, inhibit angiotensin-converting enzyme (ACE), therefore, their use in the treatment of COVID-19 patients with injury of the ACE2 expressing organs is considered. In addition, many cyanobacterial extracts were revealed to have antiviral activities, but the active agents have not been identified. This fact provides a good basis for further studies on the therapeutic potential of these microorganisms.

The Gut Microbiota of Laying Hens and Its Manipulation with Prebiotics and Probiotics To Enhance Gut Health and Food Safety.

The microbiota plays a vital role in maintaining gut health and influences the overall performance of chickens. Most gut microbiota-related studies have been performed in broilers, which have different microbial communities compared to those of layers. The normal gut microbiota of laying chickens is dominated by Proteobacteria, Firmicutes, Bacteroidetes, Fusobacteria, and Actinobacteria at the phylum level. The composition of the gut microbiota changes with chicken age, genotype, and production system. The metabolites of gut microbiota, such as short-chain fatty acids, indole, tryptamine, vitamins, and bacteriocins, are involved in host-microbiota cross talk, maintenance of barrier function, and immune homeostasis. Resident gut microbiota members also limit and control the colonization of foodborne pathogens. In-feed supplementations of prebiotics and probiotics strengthen the gut microbiota for improved host performance and colonization resistance to gut pathogens, such as Salmonella and Campylobacter The mechanisms of action of prebiotics and probiotics come through the production of organic acids, activation of the host immune system, and production of antimicrobial agents. Probiotic candidates, including Lactobacillus, Bifidobacterium, Bacillus, Saccharomyces, and Faecalibacterium isolates, have shown promising results toward enhancing food safety and gut health. Additionally, a range of complex carbohydrates, including mannose oligosaccharides, fructo-oligosaccharides, and galacto-oligosaccharides, and inulin are promising candidates for improving gut health. Here, we review the potential roles of prebiotics and probiotics in the reshaping of the gut microbiota of layer chickens to enhance gut health and food safety.

Synthesis and characterization of calcium carbonate on three kinds of microbial cells templates

As is known, the formation of nanocrystals is influenced by the cells with which they interact. In particular, the same materials could exhibit different properties depending on the type of cells. In this work, we used three different types of microbial cells (Escherichia coli, Staphylococcus aureus, and Beer Yeast) as biological templates to control the crystallization of calcium carbonate particles before or after coated with several layers of polyelectrolyte films via Layer-by-Layer assembly and biomineralization process. The results showed that the resulting crystal types using different naked cells as templates all include calcite and vaterite, and the ratio of vaterite is higher. After encapsulating with polyelectrolyte, polyelectrolyte did not change the ratio of calcite to vaterite on the surface of E. coli and Staphylococcus aureus but yeast. Furthermore, it can weaken the intensity of vaterite and increase the proportion of calcite using the yeast as template. Differences in calcium carbonate crystal forms are related to the differences in cell wall components and cell secretion. So we mineralized calcium carbonate with the main components of the yeast cell wall (mannose oligosaccharide, β-glucan) and the Gram bacterial wall (peptidoglycan, lipopolysaccharide) directly, it was found that the composition of the cell wall did affect the crystal form and morphology of the calcium carbonate. This provides a reference for further study of the biomineralization mechanism and the selection of mineralization templates. Furthermore, this study provides some supplementary for understanding the natural mineralization process of CaCO3.

Regulation of Neutrophil Function by Selective Targeting of Glycan Epitopes Expressed on the Integrin CD11b/CD18

Polymorphonuclear neutrophils (PMN) play a critical role in the innate immune response to invading pathogens. However, dysregulated mucosal trafficking of PMN and associated epithelial tissue damage is a pathological hallmark of numerous inflammatory conditions including inflammatory bowel disease and chronic obstructive pulmonary disease. The glycoprotein CD11b/CD18 plays a well described role in regulating PMN transepithelial migration and PMN inflammatory functions. Previous studies have demonstrated that targeting of the N‐linked glycan Lewis X on CD11b blocks PMN transepithelial migration (TEpM). Given evidence of glycosylation dependent regulation of CD11b/CD18 function, we performed MALDI TOF Mass Spectrometry (MS) analyses on CD11b/CD18 purified from human PMN. MS analyses revealed a surprising lack of sialylation on CD11b glycans. Unusual glycan epitopes identified on CD11b/CD18 included high Mannose oligosaccharides recognized by the Galanthus Nivalis (GNA) lectin and biantennary galactosylated N‐glycans recognized by the Phaseolus Vulgaris erythroagglutinin (PHA‐E) lectin. Importantly, we show that selective targeting of glycans on CD11b with such lectins results in altered intracellular signaling events that inhibit TEpM and differentially affect key PMN inflammatory functions including phagocytosis, superoxide release and apoptosis. Taken together, these data demonstrate that discrete glycan motifs expressed on CD11b/CD18 such as biantennary galactose could represent novel targets for selective manipulation of CD11b function and reduction of PMN associated tissue damage in chronic inflammatory diseases.Support or Funding Information This work was supported by the Crohn’s and colitis foundation (SRA # 544596)

Cloning,Expression and Characterization of Recombinant β -mannanase from Bacillus subtilis

To achieve the heterologous expression of beta-mannanase which is highly specific to Konjac glucomannan,the beta-mannanase gene BsmanA was cloned from Bacillus subtilis G1,linked to the expression vector pACYCDuet-1 and transformed into E.coil BL21(DE3). The results showed that the full length of the beta-mannanase gene BsmanA was 1098 bp and encoded 366 amino acids. After purification by Ni affinity chromatography,the molecular weight of recombinant enzyme was determined to be 38 kD. The results of enzymatic properties of recombinant beta-mannanase showed that the optimum temperature and pH for the enzymatic reaction were 60℃ and pH6.5.The stability of recombinant beta-mannanase could be maintained in the range of pH4.5~7.0 and temperature 50~70℃. In this study,the heterologous expression of beta-mannanase was accomplished,which would provide a new choice for the industrialization of biocatalytic preparation of mannose oligosaccharides.

Regulation of neutrophil function by selective targeting of glycan epitopes expressed on the integrin CD11b/CD18.

Polymorphonuclear neutrophils (PMNs) play a critical role in the innate immune response to invading pathogens. However, dysregulated mucosal trafficking of PMNs and associated epithelial tissue damage is a pathological hallmark of numerous inflammatory conditions including inflammatory bowel disease. The glycoprotein CD11b/CD18 plays a well-described role in regulating PMN transepithelial migration and PMN inflammatory functions. Previous studies have demonstrated that targeting of the N-linked glycan Lewis X on CD11b blocks PMN transepithelial migration (TEpM). Given evidence of glycosylation-dependent regulation of CD11b/CD18 function, we performed MALDI TOF Mass Spectrometry (MS) analyses on CD11b/CD18 purified from human PMNs. Unusual glycan epitopes identified on CD11b/CD18 included high Mannose oligosaccharides recognized by the Galanthus Nivalis lectin and biantennary galactosylated N-glycans recognized by the Phaseolus Vulgaris erythroagglutinin lectin. Importantly, we show that selective targeting of glycans on CD11b with such lectins results in altered intracellular signaling events that inhibit TEpM and differentially affect key PMN inflammatory functions including phagocytosis, superoxide release and apoptosis. Taken together, these data demonstrate that discrete glycan motifs expressed on CD11b/CD18 such as biantennary galactose could represent novel targets for selective manipulation of CD11b function and reduction of PMN-associated tissue damage in chronic inflammatory diseases.

Comparative binding and uptake of liposomes decorated with mannose oligosaccharides by cells expressing the mannose receptor or DC-SIGN

Mannose Receptor (MR) and DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) are two mannose-specific targets for antigens carried by liposomes but DC-SIGN is more specific of DCs. Here, DC targeting is addressed by using DPPC/DOPE liposomes decorated with a series of diether lipids with a polar head of either a mannose (Man), tri-antenna of α-d-mannopyranoside (Tri-Man), [Manα1-3(Manα1-6)Man] (Man-tri), pseudo-Man4 (PMan4) or pseudo-Man5 (PMan5). Liposomes decorated with Man-Tri show the highest binding and internalization in cells expressing DC-SIGN and in human monocytes-derived DCs. Conversely, cells expressing MR bind and take up Tri-Man liposomes 3-fold higher than Man-tri liposomes. Comparatively, liposomes decorated with PMan4 and PMan5 do not show any advantages. Overall, the results indicate that liposomes decorated with Man-tri residues are more selective toward DCs than those with Tri-Man thanks to better recognition by DC-SIGN.

Mannan oligosaccharide increases the growth performance, immunity and resistance capability against Vibro Parahemolyticus in juvenile abalone Haliotis discus hannai Ino

This trial was conducted to investigate the effect of mannose oligosaccharides (MOS) on the growth performance, antioxidation, immunity and disease resistance of Vibro Parahemolyticus in juvenile abalone Haliotis discus hannai Ino. Four formulated diets were produced to contain 0.00 g/kg, 0.40 g/kg, 0.80 g/kg and 1.60 g/kg Actigen®, with functional ingredients of MOS, respectively. Accordingly, the experimental diets were named as A0, A4, A8 and A16. After 120-days feeding trial, the best growth performance was observed in A8 group (P < 0.05) and there was no significant difference in A0, A4 and A16 groups. With the increase of dietary MOS, the activity of the total antioxidant capacity in hepatopancreas is increasingly elevated (P < 0.05) while no significant difference was observed on activity of glutathione S‐transferase (P > 0.05). The activities of superoxide dismutase and glutathione peroxidase were firstly increased and then decreased, with the highest values in A8 group (P < 0.05). Immune-related parameters were significantly affected by dietary MOS inclusion. Specifically, the activities of alkaline phosphatase and acid phosphatase in hepatopancreas and serum of abalone fed diets containing MOS were significantly higher than those of control A0 group (P < 0.05). Moreover, the highest values of both enzymes were observed in hepatopancreas of A8 group but in serum of A16 group, respectively. The lysozyme activities in hepatopancreas and serum of A4 group were significantly higher than those of other groups (P < 0.05) and there was no significant difference in A0, A8 and A16 groups (P > 0.05). The activities of cytophagy and respiratory burst in serum of abalone were not significantly affected by dietary MOS content (P > 0.05). The mRNA levels of focal adhesion kinase and integrin-linked kinase were gradually elevated with the increase of dietary MOS, with the highest value recorded in A16 group (P < 0.05). The gene expression of caspse-3 in A8 group was dramatically higher than those of other groups (P < 0.05) and there was no significant difference in A0, A4 and A16 groups (P > 0.05). The mRNA level of nuclear factor-κB was not significantly affected by dietary MOS (P > 0.05). During 56 h of V. Parahemolyticus challenge period, the accumulated mortality rate of abalone fed diets containing MOS were significantly lower than that of control A0 group in each time point (P < 0.05). Overall, the lowest rate was happened in A8 group (P < 0.05). In conclusion, MOS inclusion in diet has obviously positive effect on growth, immunity and disease resistance capability of abalone, with the optimal level of Actigen® at 0.80 g/kg in diet.

Mannan Oligosaccharide Suppresses Lipid Accumulation and Appetite in Western-Diet-Induced Obese Mice Via Reshaping Gut Microbiome and Enhancing Short-Chain Fatty Acids Production.

Obesity is associated with gut microbiome dysbiosis. Mannose oligosaccharide (MOS) has been reported to be a potential prebiotic. The present study is aimed to determine the effects of MOS on western-diet-induced obesity and to uncover the mediating roles of the gut microbiota and microbial metabolites. Three-month-old male ICR mice are fed with a high-fat and high-fructose diet for 8 weeks. The diet-induced obese mice are then orally administrated with MOS (100 and 200mgkg-1 d-1 ) for 4 weeks. MOS significantly reduces bodyweight gain, insulin resistance, fatty liver, and inflammatory responses in obese mice. MOS also stimulates lipolysis and inhibits lipogenesis in the adipose tissues. Moreover, MOS restructures the gut microbiome by enhancing the abundance of Bifidobacterium and Lactobacillus in obese mice. The microbial metabolite SCFAs are also increased in the feces and serum. Correlation analysis indicates that the appetite suppression and lipid-lowering effects of MOS are highly correlated with the butyrate levels. MOS suppresses the appetite, which results in less lipid deposition. The lower appetite is likely due to an altered gut microbiome and elevated SCFAs production. MOS may be a potential nutraceutical used in body weight management and gut health improvement.