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Background: The efficacy of allergen-specific immunotherapy (AIT) is mainly depended on the tolerogenic immune responses elicited. Properly conjugated nano-vaccine has the advantages of both specific targeting and continuous and on-demand release of allergen. Objectives: The aim of this study is to investigate the effects of a dendritic cells (DCs)-targeting nano-vaccine for AIT. Methods: The nano-vaccine was produced by coupling polylactic-co-glycolic acid (PLGA)-encapsulated ovalbumin (OVA) with mannan. Allergen capture, human monocytes-derived DCs (hMoDCs) activation, and T cells responses were assessed by flow cytometry, confocal microscopy, quantitative real-time PCR, ELISA, and Cytometric Bead Array. Balb/c mice were immunized with the nano-vaccines, and the immune responses were analyzed. Results: OVA-PLGA nanoparticle (NP) displayed favorable safety profile. OVA-mannan-PLGA NP was captured more efficiently by hMoDCs than OVA-PLGA NP, which was mediated mainly through DC-specific intercellular adhesion molecule 3-grabbing nonintegrin. A tolerogenic phenotype of hMoDCs was induced by OVA-mannan-PLGA NP, but not OVA-PLGA NP, and increased number of regulatory T (Treg) cells was generated subsequently in in vitro coculture. Immunization of Balb/c mice with OVA-mannan-PLGA NP resulted in lower serum level of OVA-specific immunoglobulins and less production of pro-inflammatory cytokines in splenocytes culture than the mice immunized with OVA-PLAG NP, PLGA NP, or OVA, while the number of splenic Treg cells was higher in OVA-mannan-PLGA group than in other groups. Moreover, preimmunization with OVA-mannan-PLGA NP significantly inhibited the Th2 immune response induced by OVA sensitization. Conclusions: The biocompatible PLGA-encapsulated OVA coupling with mannan has augmented ability for tolerance induction and could be developed as a novel vaccine for AIT. [ABSTRACT FROM AUTHOR]

Diagnosing invasive or chronic fungal infections is a challenge, particularly in the immunocompromised host. Microscopy and culture remain the reference standard, but are insensitive. The use of non-culture-based techniques is recommended in conjunction with conventional methods to improve the diagnostic yield. The aim was to provide an updated 2021 inventory of fungal antigen and serology tests for diagnosing invasive and chronic fungal infections, the key focus was set on Aspergillus , Candida and Cryptococcus species. Pubmed search for publications with the key words fungal antigen tests, laboratory-based diagnosis of invasive pulmonary aspergillosis, chronic pulmonary aspergillosis, invasive candidiasis, invasive fungal infections and cryptococcal infections published from 2017 to 2020. Antigen assays such as the galactomannan (GM) and β- d -glucan detection systems are frequently used, but these tests vary in sensitivity and specificity, depending on the patient population involved, specimens inspected, cut-offs defined, test strategy applied and inclusion or exclusion of possible fungal case definitions. Multiple different detection systems are available, with recently introduced new point-of-care tests such as the lateral flow device and the lateral flow assay. Despite a wide heterogeneity in populations evaluated, studies indicate a better diagnostic performance of bronchoalveolar lavage GM in comparison with serum GM, and a suboptimal specificity of GM bronchoalveolar lavages (cut-off ≥1) and serum β- d -glucan in non-neutropenic individuals. Point-of-care cryptococcal antigen tests show excellent performance. There are fungal antigen detection tests available with excellent to reasonable clinical performance to diagnose invasive fungal infections. Only a few assays are useful to monitor therapeutic response. There are multiple marketed IgG antibody tests to detect Aspergillus fumigatus antibodies, the titres vary widely and the performance differs significantly. In general, diagnostic tests are vulnerable to being affected by the host, the microbe and laboratory setting. [ABSTRACT FROM AUTHOR]

Nanoparticle-based cancer immunotherapy is considered a novel and promising therapeutic strategy aimed at stimulating host immune responses against tumors. To this end, in the present study, mannan-decorated polylactic-co-glycolic acid (PLGA) nanoparticles containing tumor cell lysate (TCL) and poly riboinosinic polycytidylic acid (poly I:C) were used as antigen delivery systems to immunize breast tumor-bearing Balb/c mice. PLGA nanoparticles were fabricated employing a double emulsion solvent evaporation method. The formation of spherical and uniform nanoparticles (NPs) ranging 150–250 nm was detected by field emission scanning electron microscopy (FESEM) and dynamic light scattering (DLS). Four nanoformulation were used to treat mice and vaccination-induced immunological responses. Tumor regression and overall survival rate were evaluated in four experimental groups. Tumor cell lysate and poly I:C loaded mannan-decorated nanoparticles (TCL-Poly I:C) NP-MN caused a significant decrease in tumor growth and 2- to 3-fold improvement in survival times of the treated mice. The NPs with or without mannan decoration elicited stronger responses in terms of lymphocyte proliferation, delayed-type hypersensitivity and CD107a expression. Moreover, our data indicated that the production of IFN-γ and IL-2 increased while the production of IL-4 and IL-10 decreased in splenocytes culture supernatants. In the pathological evaluations, we found that necrosis and immune cells infiltration rate in the tumor tissue of the treated mice was elevated, while tumor cellularity and lung metastases significantly decreased in particular in the group that received (TCL-Poly I:C) NP-MN. Altogether, our findings suggested that the mannan-decorated PLGA NPs antigen delivery system had significant anti-tumor effects against the murine model of breast cancer and it could be considered as a step forward to human breast cancer immunotherapy.

Bacillus Calmette-Guérin polysaccharide and nucleic acid (BCG-PSN), extracted from Mycobacterium bovis, is an immunoregulatory medicine commonly used in clinic. However, the structural characteristics and potential pharmacological efficacy of the polysaccharides from BCG-PSN remain unclear. Herein, two polysaccharides (BCG-1 and BCG-2) were purified and their structures were characterized. Monosaccharide composition analysis combined with methylation analysis and NMR data indicated that BCG-1 and BCG-2 were an α-D-(1→4)-mannan with (1→2)-linked branches, and an α-D-(1→4)-glucan with (1→6)-linked branches, respectively. Herein, the mannan from BCG-PSN was first reported. Bioactivity assays showed that BCG-1 and BCG-2 dose-dependently and potently increased the production of inflammatory mediators (NO, TNF-α, IL-6, IL-1β, and IL-10), as well as their mRNA expressions in RAW264.7 cells; both have similar or stronger effects compared with BCG-PSN injection. These data suggest that BCG-1 and BCG-2 are very likely the active ingredients of BCG-PSN.

Monoclonal antibody EBCA-1 is used in the sandwich immune assay for the detection of circulating Candida mannan in blood sera samples for the diagnosis of invasive candidiasis. To reinvestigate carbohydrate specificity of EBCA-1, a panel of biotinylated oligosaccharides structurally related to distinct fragments of Candida mannan were loaded onto a streptavidin-coated plate to form a glycoarray. Its use demonstrated that EBCA-1 recognizes the trisaccharide β-Man-(1→2)-α-Man- (1→2)-α-Man and not homo-α-(1→2)-linked pentamannoside, as was reported previously. [ABSTRACT FROM AUTHOR]

Açaí residual biomass can be a potential source for a wide range of applications, especially biotechnological, such as solid-state fermentation processes and production of fermentable sugars (mannose), such as raw material for food, pharmaceutical, and other industries. Fiber and seed of açaí fruit (Euterpe oleracea) were characterized, after pulp extraction, in samples collected at different maturation stages (seasonality). The seed contained a remarkable amount of mannose (75%) followed by glucose (6%) and galactose (2%), with mannan as the main polysaccharide. The fiber was a lignocellulosic biomass containing glucose (30%) and xylose (19%). Mannan and cellulose in the seed and fiber, respectively, were the polysaccharide responsible for the crystallinity of both materials, with the seed showing higher values. The period of fruit harvesting (winter or summer crops) influenced the maturity and chemical composition of the residues, which had higher crystallinity in the summer season. Hydrothermal treatment was very efficient for fiber but failed for seed, indicating a high recalcitrance of mannan in açaí seed. [ABSTRACT FROM AUTHOR]

Anti-viral and anti-tumor vaccines aim to induce cytotoxic CD8+ T cells (CTL) and antibodies. Conserved protein antigens, such as p24 from human immunodeficiency virus, represent promising component for elicitation CTLs, nevertheless with suboptimal immunogenicity, if formulated as recombinant protein. To enhance immunogenicity and CTL response, recombinant proteins may be targeted to dendritic cells (DC) for cross presentation on MHCI, where mannose receptor and/or other lectin receptors could play an important role. Here, we constructed liposomal carrier-based vaccine composed of recombinant p24 antigen bound by metallochelating linkage onto surface of nanoliposomes with surface mannans coupled by aminooxy ligation. Generated mannosylated proteonanoliposomes were analyzed by dynamic light scattering, isothermal titration, and electron microscopy. Using murine DC line MutuDC and murine bone marrow derived DC (BMDC) we evaluated their immunogenicity and immunomodulatory activity. We show that p24 mannosylated proteonanoliposomes activate DC for enhanced MHCI, MHCII and CD40, CD80, and CD86 surface expression both on MutuDC and BMDC. p24 mannosylated liposomes were internalized by MutuDC with p24 intracellular localization within 1 to 3 h. The combination of metallochelating and aminooxy ligation could be used simultaneously to generate nanoliposomal adjuvanted recombinant protein-based vaccines versatile for combination of recombinant antigens relevant for antibody and CTL elicitation. [Display omitted] [ABSTRACT FROM AUTHOR]

Ganoderma lucidum , widely used in traditional medicine, has several biological properties. Polysaccharides, mainly glucans, are known as one of its main bioactive compounds. Consequently, the achievement and chemical investigation of such molecules are of pharmaceutical interest. Herein, we obtained water-insoluble and water-soluble polysaccharides from G. lucidum by alkaline extraction. Fractionation process yielded three fractions (GLC-1, GLC-2, and GLC-3). All samples showed to be composed mainly of glucans. GLC-1 is a linear (1 → 3)-linked β-glucan; GLC-2 is a mixture of three different linear polysaccharides: (1 → 3)-β-glucan, (1 → 3)-α-glucan, and (1 → 4)-α-mannan; while GLC-3 is a branched β-glucan with a (1 → 4)-linked main chain, which is branched at O-3 or O-6 by (1 → 3)- or (1 → 6)-linked side chains. This research reports the variability of glucans in Ganoderma lucidum fruiting bodies and applicable methodologies to obtain such molecules. These polysaccharides can be further applied in biological studies aiming to investigate how their chemical differences may affect their biological properties. [Display omitted] • A branched glucan, linear glucans and mannan were obtained from Ganoderma lucidum. • GLC-1 is a linear (1. → 3)-linked β-glucan. • GLC-2 is a mixture of: (1. → 3)-β-glucan, (1 → 3)-α-glucan, and (1 → 4)-α-mannan. • GLC-3 is a branched β-glucan with a (1. → 4), (1 → 3), and (1 → 6) linkage types. [ABSTRACT FROM AUTHOR]

Synovial fluids from rheumatoid and psoriatic arthritis patients have high levels of PLA1A. The current study was to understand PLA1A functions in the pathophysiology of rheumatic diseases. We generated Pla1a−/− mice to assess their phenotype and the impact of PLA1A deficiency on the development of mannan-induced psoriatic arthritis (MIP). Mice were evaluated routinely for the induced symptoms. On the day of sacrifice, blood samples were collected for hematology analysis and prepared for plasma. Livers were collected. Lymph node immune cells were analyzed using flow cytometry. We performed μCT scans of hind paws from naïve and mannan-induced female mice. Cytokines/chemokines were quantified using Luminex in hind paw tissues and plasma of female mice. Pla1a−/− mice showed a slight increase in circulating and lymph node lymphocytes. CD4+ T cells contributed most to this increase in lymph nodes (p = 0.023). In the MIP model, the lymph node ratios of CD3+ to CD19+ and CD4+ to CD8+ were higher in Pla1a−/− mice. Pla1a−/− mice were less susceptible to MIP (p < 0.001) and showed reduced bone erosions. Pla1a−/− mice also showed reduced IL-17, KC, IP-10, MIP-1β, LIF, and VEGF in hind paw tissues as compared to WT mice (p < 0.05). These findings indicated that PLA1A deficiency protected from the development of the MIP disease. The data suggested that PLA1A could contribute to MIP through increased activation of lymphocytes, possibly those producing IL-17.

Polysaccharides were extracted from natural sources with various biological activities, which are strongly influenced by their chemical structure and molecular weight. In this research, mannans polysaccharides were obtained from Saccharomyces cerevisiae by ethanol precipitation. The molecular weight of YM50, YM70, and YM90 mannans was 172.90 kDa, 87.09 kDa, and 54.05 kDa, respectively. Scanning electron microscopy of YM 90 mannans showed a rough surface with numerous cavities, while the surfaces of YM50 and YM70 were relatively smooth. Sepharose CL-6B and FTIR indicated that mannans had the characteristic bands of polysaccharides. The antioxidant activities of polysaccharides were evaluated in vitro using various assays. Mannans showed a good scavenging activity of DPPH radicals which depend on the molecular weight and concentration, and a higher scavenging activity of hydroxyl radical than ferric-reducing power activities. For the three types of mannans, cytotoxicity and hemolytic activity were rarely detected in mice erythrocytes and Caco-2 cells. Those results could contribute to the further application of mannans from Saccharomyces cerevisiae in the food and medicine industry.

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Within the midgut gland of the Christmas Island red crab, Gecarcoidea natalis, a single transcript for a GH5_10 endo-β-1,4-mannase had the highest expression out of all of the carbohydrase enzymes (Gan et al. in Mar Biotechnol 20:654–665, 2018). The activity, and potential digestive importance of this hemicellulase, compared with other carbohydrases, has yet to be established. The digestive fluid of G. natalis contained substantial endo-β-1,4-mannase activities (630 ± 55 (6) nmol reducing sugars. min−1. mg−1 protein). It was present as a single isozyme of 66.3 ± 0.7 kDa (n = 6). Endo-β-1,4-mannase activities were higher than that for lichenase and endo-β-1,4-glucanase but lower than that for β-1,3-glucanase and amylase. The digestive fluid was able to hydrolyse, galactomannan, into its component monosaccharides. Hence, this confirms expression data that this enzyme is one of the most important digestive cellulases/ hemicellulases. Expression of GH5_10 endo-β-1,4-mannase mRNA was consistent with that of a digestive enzyme, as it was expressed in the digestive midgut gland but not in muscle and gill. Endo-β-1,4-mannase activities were also present within the digestive fluid of the terrestrial hermit crabs, Coenobita perlatus and Coenobita brevimanus. Endo-β-1,4-mannase activities (1351 ± 136 (n=3) nmol reducing sugars. min−1 mg−1 protein for C. perlatus. 665 ± 32 n=(5) nmol reducing sugars. min−1 mg−1 protein for C. brevimanus) were higher than that for endo-β-1,4-glucanase and amylase but were lower than β-1,3-glucanase activities. Animals within the terrestrial hermit crab family, Coenobitidae consume legume and palm seeds which contain substantial amounts of mannan. Hence, high endo-β-1,4-mannase activities suggest that digestion of mannan within these species may represent an important source of carbohydrate. [ABSTRACT FROM AUTHOR]

Candida species, belonging to commensal microbial communities in humans, cause opportunistic infections in individuals with impaired immunity. Pathogens encountered in more than 90% cases of invasive candidiasis include C. albicans, C. glabrata, C. krusei, C. tropicalis, and C. parapsilosis. The most frequently diagnosed invasive infection is candidemia. About 50% of candidemia cases result in deep-seated infection due to hematogenous spread. The sensitivity of blood cultures in autopsy-proven invasive candidiasis ranges from 21% to 71%. Non-cultural methods (beta-D-glucan, T2Candida assays), especially beta-D-glucan in combination with procalcitonin, appear promising in the exclusion of invasive candidiasis with high sensitivity (98%) and negative predictive value (95%). There is currently a clear deficiency in approved sensitive and precise diagnostic techniques. Omics technologies seem promising, though require further development and study. Therapeutic options for invasive candidiasis are generally limited to four classes of systemic antifungals (polyenes, antimetabolite 5-fluorocytosine, azoles, echinocandins) with the two latter being highly effective and well-tolerated and hence the most widely used. Principles and methods of treatment are discussed in this review. The emergence of pan-drug-resistant C. auris strains indicates an insufficient choice of available medications. Further surveillance, alongside the development of diagnostic and therapeutic methods, is essential.

Carbohydrate-binding modules (CBMs) are usually appended to carbohydrate-active enzymes (CAZymes) and serve to potentiate catalytic activity, for example, by increasing substrate affinity. The Gram-negative soil saprophyte Cellvibrio japonicus is a valuable source for CAZyme and CBM discovery and characterization due to its innate ability to degrade a wide array of plant polysaccharides. Bioinformatic analysis of the CJA_2959 gene product from C. japonicus revealed a modular architecture consisting of a fibronectin type III (Fn3) module, a cryptic module of unknown function (X181), and a glycoside hydrolase family 5 subfamily 4 (GH5_4) catalytic module. We previously demonstrated that the last of these, CjGH5F, is an efficient and specific endo-xyloglucanase (M. A. Attia, C. E. Nelson, W. A. Offen, N. Jain, et al., Biotechnol Biofuels 11:45, 2018, https://doi.org/10.1186/s13068-018-1039-6). In the present study, C-terminal fusion of superfolder green fluorescent protein in tandem with the Fn3-X181 modules enabled recombinant production and purification from Escherichia coli. Native affinity gel electrophoresis revealed binding specificity for the terminal galactose-containing plant polysaccharides galactoxyloglucan and galactomannan. Isothermal titration calorimetry further evidenced a preference for galactoxyloglucan polysaccharide over short oligosaccharides comprising the limit-digest products of CjGH5F. Thus, our results identify the X181 module as the defining member of a new CBM family, CBM88. In addition to directly revealing the function of this CBM in the context of xyloglucan metabolism by C. japonicus, this study will guide future bioinformatic and functional analyses across microbial (meta)genomes. [ABSTRACT FROM AUTHOR]

Primary fungal infection of the central nervous system (CNS) is rare but often associated with severe prognosis. Diagnosis is complicated since cerebrospinal fluid (CSF) samples obtained from lumbar puncture usually remain sterile. Testing for fungal antigens in CSF could be a complementary diagnostic tool. We conducted such measurements in CSF from patients with CNS fungal infection and now discuss the usefulness of ventricular puncture. Mannan and (1→3)ß-D-glucan (BDG) testing were retrospectively performed in CSF samples from three patients with proven chronic CNS fungal infection (excluding Cryptococcus), and subsequently compared to 16 controls. Results from lumbar punctures and those from cerebral ventricles were confronted. BDG detection was positive in all the CSF samples (from lumbar and/or ventricular puncture) from the three confirmed cases. In case of Candida infection, mannan antigen measurement was positive in 75% of the CSF samples. In the control group, all antigen detections were negative (n  = 15), except for one false positive. Faced with suspected chronic CNS fungal infection, measurement of BDG levels appears to be a complementary diagnostic tool to circumvent the limitations of mycological cultures from lumbar punctures. In the event of negative results, more invasive procedures should be considered, such as ventricular puncture. [ABSTRACT FROM AUTHOR]

The cellular surface of the pathogenic filamentous fungus Aspergillus fumigatus is enveloped in a mannose layer, featuring well-established fungal-type galactomannan and O -mannose-type galactomannan. This study reports the discovery of cell wall component in A. fumigatus mycelium, which resembles N -glycan outer chains found in yeast. The glycosyltransferases involved in its biosynthesis in A. fumigatus were identified, with a focus on two key α-(1→2)-mannosyltransferases, Mnn2 and Mnn5, and two α-(1→6)-mannosyltransferases, Mnn9 and Van1. In vitro examination revealed the roles of recombinant Mnn2 and Mnn5 in transferring α-(1→2)-mannosyl residues. Proton nuclear magnetic resonance ( 1 H-NMR) analysis of cell wall extracts from the ∆ mnn2 ∆ mnn5 strain indicated the existence of an α-(1→6)-linked mannan backbone in the A. fumigatus mycelium, with Mnn2 and Mnn5 adding α-(1→2)-mannosyl residues to this backbone. The α-(1→6)-linked mannan backbone was absent in strains where mnn9 or van1 was disrupted in the parental ∆ mnn2 ∆ mnn5 strain in A. fumigatus . Mnn9 and Van1 functioned as α-(1→6)-linked mannan polymerases in heterodimers when co-expressed in Escherichia coli , indicating their crucial role in biosynthesizing the α-(1→6)-linked mannan backbone. Disruptions of these mannosyltransferases did not affect fungal-type galactomannan biosynthesis. This study provides insights into the complexity of fungal cell wall architecture and a better understanding of mannan biosynthesis in A. fumigatus ., Importance: This study unravels the complexities of mannan biosynthesis in A. fumigatus , a key area for antifungal drug discovery. It reveals the presence of α-(1→6)-linked mannan structures resembling yeast N-glycan outer chains in A. fumigatus mycelium, offering fresh insights into the fungal cell wall's design. Key enzymes, Mnn2, Mnn5, Mnn9, and Van1, are instrumental in this process, with Mnn2 and Mnn5 adding specific mannose residues and Mnn9 and Van1 assembling the α-(1→6)-linked mannan structures. Although fungal-type galactomannan's presence in the cell wall is known, the existence of an α-(1→6)-linked mannan adds a new dimension to our understanding. This intricate web of mannan biosynthesis opens avenues for further exploration and enhances our understanding of fungal cell wall dynamics, paving the way for targeted drug development., Competing Interests: The authors declare no conflict of interest.

Poly(methyl methacrylate) (PMMA) is commonly used for dental dentures, but it has the drawback of promoting oral health risks due to oral bacterial adhesion. Recently, various nanoparticles have been incorporated into PMMA to tackle these issues. This study aims to investigate the mechanophysical and antimicrobial adhesive properties of a denture resin by incorporating of nanoclay into PMMA. Specimens were prepared by adding 0, 1, 2, and 4 wt % surface-modified nanoclay (Sigma) to self-polymerizing PMMA denture resin. These specimens were then evaluated using FTIR, TGA/DTG, and FE-SEM with EDS. Various mechanical and surface physical properties, including nanoindentation, were measured and compared with those of pure PMMA. Antiadhesion experiments were conducted by applying a Candida albicans (ATCC 11006) suspension to the surface of the specimens. The antiadhesion activity of C. albicans was confirmed through a yeast-wall component (mannan) and mRNA-seq analysis. The bulk mechanical properties of nanoclay-PMMA composites were decreased compared to those of pure PMMA, while the flexural strength and modulus met the ISO 20795-1 requirement. However, there were no significant differences in the nanoindentation hardness and elastic modulus. The surface energy revealed a significant decrease at 4 wt % nanoclay-PMMA. The antiadhesion effect of Candida albicans was evident along with nanoclay content in the nanocomposites and confirmed by the reduced attachment of mannan on nanoclay-PMMA composites. mRNA-seq analysis supported overall transcriptome changes in altering attachment and metabolism behaviors on the surface. The nanoclay-PMMA materials showed a lower surface energy as the content increased, leading to an antiadhesion effect against Candida albicans . These findings indicate that incorporating nanoclay into PMMA surfaces could be a valuable strategy for preventing the fungal biofilm formation of denture base materials.

Background: Chemo-immunotherapy modifies the tumor microenvironment to enhance the immune response and improve chemotherapy. This study introduces a dual-armed chemo-immunotherapy strategy combating breast tumor progression while re-polarizing Tumor-Associated Macrophage (TAM) using prodigiosin-loaded mannan-coated magnetic nanoparticles (PG@M-MNPs)., Methods: The physicochemical properties of one-step synthetized M-MNPs were analyzed, including X-ray diffraction, FTIR, DLS, VSM, TEM, zeta potential analysis, and drug loading content were carried out. Biocompatibility, cancer specificity, cellular uptake, and distribution of PG@M-MNPs were investigated using fluorescence and confocal laser scanning microscopy, and flow cytometry. Furthermore, the expression levels of IL-6 and ARG-1 after treatment with PG and PG@M-MNPs on M1 and M2 macrophage subsets were studied., Results: The M-MNPs were successfully synthesized and characterized, demonstrating a size below 100 nm. The release kinetics of PG from M-MNPs showed sustained and controlled patterns, with enzyme-triggered release. Cytotoxicity assessments revealed an enhanced selectivity of PG@M-MNPs against cancer cells and minimal effects on normal cells. Additionally, immuno-modulatory activity demonstrates the potential of PG@M-MNPs to change the polarization dynamics of macrophages., Conclusion: These findings highlight the potential of a targeted approach to breast cancer treatment, offering new avenues for improved therapeutic outcomes and patient survival.

Mannan and outer structural yeast cell wall polysaccharides have recently garnered attention for their health defense and cosmetic applications. In addition, many studies have confirmed that yeast cell wall mannans exhibit various biological activities, such as antioxidant, immune regulation, reducing hyperlipidemia, and gut health promotion. This paper elucidates yeast cell wall mannan structural features, biological activities, underlying molecular mechanisms, and biosynthesis. Moreover, mannan-overproducing strategies through yeast strain engineering are emphasized and discussed. This review will provide a scientific basis for yeast cell wall mannan research and industrial applications., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Sanghuang is a well-known edible and medicinal fungus consumed in Asia for centuries. In the present study, a novel exopolysaccharide named as mannan (SSEPS2) was isolated from the liquid culture broth of Sanghuangporus sanghuang by using ethanol precipitation, anion-exchange chromatography, and size-exclusion chromatography, which was exclusively composed of D-mannose with the molecular weight 9.43 × 104 g/mol. The structural analysis suggested that SSEPS2 consisted of 1,3-linked and 1,2-linked α-D-mannopyranose (Man p), with substitution at O -6 of 1,2-linked α-D-Man p by 1,6-linked α-D-Man p residues and terminal α-D-Man p residues. The exponent of α (0.58) indicated that SSEPS2 existed in a flexible chain conformation, with a coil-like structure in 0.1 M NaNO 3 at 25 °C, which was further confirmed by the atomic force microscopy images and a three-dimensional molecular model. It exhibited potential antitumor activity against the growth of HepG2 (human hepatoma) and MCF-7 cell lines in vitro. The IC 50 values for inhibiting HepG-2 cells and MCF-7 cells proliferation were 365 and 340 μg/mL, which showed its potential application in the development of anticancer therapeutics. • A novel mannan purified from liquid broth of Sanghuangporus sanghuang • Its backbone consisted of 1,3-linked α-D-Man p and 1,2-linked α-D-Man p. • It has a flexible chain conformation displayed antitumor activities. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is the most common trichothecene distributed in food and feed. So far, much work has focused on investigating the cytotoxicity of DON, while there is few researches aimed at intervening in the toxic impacts on humans and livestock posed by DON. The objective of this study is to investigate the underlying mechanism of biomacromolecules mannan/β-glucans from yeast cell wall (BYCW) for their potency to impede the cytotoxicity and apoptosis caused by DON with porcine jejunum epithelial cell lines (IPEC-J2) used as a cell injury model. We analyzed the cell morphology, cell activity, oxidative stress, fluorescence intensity and expressions of proteins relevant to autophagy, apoptosis and PI3K-AKT-mTOR signaling pathway by using inverted microscopy, MTS, reactive oxygen species (ROS), glutathione (GSH) and malondialdehyde (MDA) assay, Annexin V-FITC / propidium iodide (PI) double staining and Western blot assay. The consequent data demonstrated that in the presence of BYCW, the cell morphology and activity were relatively ameliorated and that the oxidation damage was attenuated with DON-induced autophagy concomitantly decreased, which, furthermore, was found involved in the positive regulation on PI3K-AKT-mTOR signaling pathway by BYCW. In a word, BYCW possess an ability to repress the cytotoxicity and apoptosis induced by DON through the inhibition of autophagy via activating PI3K-AKT-mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

Cell wall thickening and development of secondary cell walls was a major step in plant terrestrialization that provided the mechanical support, effective functioning of water-conducting elements and fortification of the surface tissues. Despite its importance, the diversity, emergence and evolution of secondary cell walls in early land plants have been characterized quite poorly. Secondary cell walls can be present in different cell types with fibers being among the major ones. The necessity for mechanical support upon increasing plant height is widely recognized; however, identification of fibers in land plants of early taxa is quite limited. In an effort to partially fill this gap, we studied the fibers and the composition of cell walls in stems of the sporophyte of the living fossil Psilotum nudum. Various types of light microscopy, combined with partial tissue maceration demonstrated that this perennial, rootless, fern-like vascular plant, has abundant fibers located in the middle cortex. Extensive immunodetection of cell wall polymers together with various staining and monosaccharide analysis of cell wall constituents revealed that in P. nudum, the secondary cell wall of its cortical fibers is distinct from that of its tracheids. Primary cell walls of all tissues in P. nudum shoots are based on mannan, which is also common in other extant early land plants. Besides, the primary cell wall contains epitope for LM15 specific for xyloglucan and JIM7 that binds methylesterified homogalacturonans, two polymers common in the primary cell walls of higher plants. Xylan and lignin were detected as the major polymers in the secondary cell walls of P. nudum tracheids. However, the secondary cell wall in its cortical fibers is quite similar to their primary cell walls, i.e., enriched in mannan. The innermost secondary cell wall layer of its fibers but not its tracheids has epitope to bind the LM15, LM6, and LM5 antibodies recognizing, respectively, xyloglucan, arabinan and galactan. Together, our data provide the first description of a mannan-based cell wall in sclerenchyma fibers, and demonstrate in detail that the composition and structure of secondary cell wall in early land plants are not uniform in different tissues.

Apart from its role as a digestive and absorptive organ, the gastrointestinal (GI) tract is a vital immune organ that encompasses roughly 70 % of the total immune cells of the body. As such, the physical, chemical and nutrient composition of the diet influences overall GI function, effectively as an immune organ. With the improvement in feed technology, agro-industrial co-products that are high in fibre have been widely used as a feed ingredient in the diets of pigs and poultry. Arabinoxylan (AX) and mannan are the most abundant hemicellulosic polysaccharides present in cereal grain and co-product ingredients used in the livestock industry. When monogastric animals consume diets containing high amounts of AX and mannans, stimulation of GI immune cells may occur. This involves the activation of several cellular and molecular pathways of the immune system and requires a considerable amount of energy and nutrients to be expended by the animal, which may ultimately influence overall health and growth performance of animals. Therefore, a better understanding of the role of AX and mannan in immune modulation will be helpful in modulating untoward GI immune responses, thereby minimising nutrient and energy expenditure toward this effort. This review will summarise pertinent research on the role of oligosaccharides and polysaccharides containing AX and mannans in immune modulation in order to preserve gut integrity.

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.

Abstract Background Acetylated galactoglucomannan (AcGGM) is a complex hemicellulose found in softwoods such as Norway spruce (Picea abies). AcGGM has a large potential as a biorefinery feedstock and source of oligosaccharides for high-value industrial applications. Steam explosion is an effective method for extraction of carbohydrates from plant biomass. Increasing the reaction pH reduces the combined severity ($$R^{\prime}_{0}$$ R0′ ) of treatment, affecting yields and properties of extracted oligosaccharides. In this study, steam explosion was used to extract oligosaccharides from Norway spruce wood chips soaked with sodium citrate and potassium phosphate buffers with pH of 4.0–7.0. Yields, monosaccharide composition of released oligosaccharides and biomass residue, their acetate content and composition of their lignin fraction were examined to determine the impact of steam explosion buffering on the extraction of softwood hemicellulose. Results Reducing the severity of steam explosion resulted in lower yields, although the extracted oligosaccharides had a higher degree of polymerization. Higher buffering pH also resulted in a higher fraction of xylan in the extracted oligos. Oligosaccharides extracted in buffers of pH > 5.0 were deacetylated. Buffering leads to a removal of acetylations from both the extracted oligosaccharides and the hemicellulose in the residual biomass. Treatment of the residual biomass with a GH5 family mannanase from Aspergillus nidulans was not able to improve the AcGGM yields. No hydroxymethylfurfural formation, a decomposition product from hexoses, was observed in samples soaked with buffers at pH higher than 4.0. Conclusions Buffering the steam explosion reactions proved to be an effective way to reduce the combined severity ($$R^{\prime}_{0}$$ R0′ ) and produce a wide range of products from the same feedstock at the same physical conditions. The results highlight the impact of chemical autohydrolysis of hemicellulose by acetic acid released from the biomass in hydrothermal pretreatments. Lower combined severity results in products with a lower degree of acetylation of both the extracted oligosaccharides and residual biomass. Decrease in severity appears not to be the result of reduced acetate release, but rather a result of inhibited autohydrolysis by the released acetate. Based on the results presented, the optimal soaking pH for fine-tuning properties of extracted AcGGM is below 5.0.

Myelin peptide–mannan conjugates have been shown to be potential vaccines in the immunotherapy of multiple sclerosis. The conjugates are comprised from the epitope peptide and the polysaccharide mannan which transfers as a carrier the antigenic peptide to dendritic cells that process and present antigenic peptides at their surface in complex with MHC class I or class II resulting in T-cell stimulation. The conjugation of antigenic peptide with mannan occurs through the linker (Lys–Gly)5, which connects the peptide with the oxidized mannose units of mannan. This study describes novel methods for the quantification of the vaccine ingredient peptide within the conjugate, a prerequisite for approval of clinical trials in the pursuit of multiple sclerosis therapeutics. Myelin peptides, such as MOG35–55, MBP83–99, and PLP131–145 in linear or cyclic form, as altered peptide ligands or conjugated to appropriate carriers, possess immunomodulatory properties in experimental models and are potential candidates for clinical trials.

Copyright of China Brewing is the property of China Brewing Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The secretory pathway in Candida albicans involves the protein translocation into the lumen of the endoplasmic reticulum and transport to the Golgi complex, where proteins undergo posttranslational modifications, including glycosylation and proteolysis. The Golgi-resident Kex2 protease is involved in such processing and disruption of its encoding gene affected virulence and dimorphism. These previous studies were performed using cells without URA3 or with URA3 ectopically placed into the KEX2 locus. Since these conditions are known to affect the cellular fitness and the host–fungus interaction, here we generated a kex2Δ null mutant strain with URA3 placed into the neutral locus RPS1. The characterization of this strain showed defects in the cell wall composition, with a reduction in the N-linked mannan content, and the increment in the levels of O-linked mannans, chitin, and β-glucans. The defects in the mannan content are likely linked to changes in Golgi-resident enzymes, as the α-1,2-mannosyltransferase and α-1,6-mannosyltransferase activities were incremented and reduced, respectively. The mutant cells also showed reduced ability to stimulate cytokine production and phagocytosis by human mononuclear cells and macrophages, respectively. Collectively, these data showed that loss of Kex2 affected the cell wall composition, the protein glycosylation pathways, and interaction with innate immune cells. [ABSTRACT FROM AUTHOR]

Colloidal stability of beer is one of the most critical challenges facing the brewing industry. In brewing, stabilization agents are widely used to remove colloids such as proteins and polyphenols. However, the removal efficiency of polyphenols depends on the yeast, which adsorbs these compounds on its surface. The adsorption of polyphenols on the yeast surface is associated with the zeta potential of the cell wall, which correlates with the mannan content in them. The purpose of the study was to investigate the role of yeast strains in adsorption of polyphenols (increasing colloidal stability of beer) and the correlation of this property with the mannan content in the cell wall during the fermentation. Two S. cerevisiae strains were used in this study, and the specific growth rate of yeast, the specific rate of change in the polyphenol content and the mannan content were determined. The results of this study showed that the highest mannan content in yeast was after 9 hours (the end of exponential growth phase). Its content was 10.97% by weight of dry matter in the yeast S. cerevisiae Californian Lager (M54), and 13.69% in the yeast S. cerevisiae Belgian Wit (M21). The desorption of polyphenols was observed during the period from 9 to 24 hours of fermentation, where an increase in the content of polyphenols in the medium was observed. Based on these results, it is necessary to remove the yeast at the end of the exponential growth phase to increase the colloidal stability of the beer. [ABSTRACT FROM AUTHOR]

Cell wall thickening and development of secondary cell walls was a major step in plant terrestrialization that provided the mechanical support, effective functioning of water-conducting elements and fortification of the surface tissues. Despite its importance, the diversity, emergence and evolution of secondary cell walls in early land plants have been characterized quite poorly. Secondary cell walls can be present in different cell types with fibers being among the major ones. The necessity for mechanical support upon increasing plant height is widely recognized; however, identification of fibers in land plants of early taxa is quite limited. In an effort to partially fill this gap, we studied the fibers and the composition of cell walls in stems of the sporophyte of the living fossil Psilotum nudum. Various types of light microscopy, combined with partial tissue maceration demonstrated that this perennial, rootless, fern-like vascular plant, has abundant fibers located in the middle cortex. Extensive immunodetection of cell wall polymers together with various staining and monosaccharide analysis of cell wall constituents revealed that in P. nudum , the secondary cell wall of its cortical fibers is distinct from that of its tracheids. Primary cell walls of all tissues in P. nudum shoots are based on mannan, which is also common in other extant early land plants. Besides, the primary cell wall contains epitope for LM15 specific for xyloglucan and JIM7 that binds methylesterified homogalacturonans, two polymers common in the primary cell walls of higher plants. Xylan and lignin were detected as the major polymers in the secondary cell walls of P. nudum tracheids. However, the secondary cell wall in its cortical fibers is quite similar to their primary cell walls, i.e., enriched in mannan. The innermost secondary cell wall layer of its fibers but not its tracheids has epitope to bind the LM15, LM6, and LM5 antibodies recognizing, respectively, xyloglucan, arabinan and galactan. Together, our data provide the first description of a mannan-based cell wall in sclerenchyma fibers, and demonstrate in detail that the composition and structure of secondary cell wall in early land plants are not uniform in different tissues. [ABSTRACT FROM AUTHOR]

Mannooligosaccharides (MOS) were derived using Aspergillus oryzae β-mannanase (ManAo) from different mannan-rich agro-wastes, palm kernel cake (PKC), guar gum and copra meal (CM). Guar gum (GG) released higher amount of MOS (56.31% w/w) from which purification of mannobiose (0.68 mg) and mannotriose (1.26 mg) was demonstrated using size-exclusion chromatography. FTIR analysis of mannan hydrolysates showed characteristic peaks in 1200–900 cm−1 region indicating the presence of MOS. 1H &13C NMR spectra showed presence of anomeric sugar forms of MOS in different mannan hydrolysates. MOS from locust bean gum and guar gum had both α- and β-anomers while PKC and CM had only α-anomer. Growth promotional activities of different MOS were demonstrated using two probiotic Lactobacilli. Besides, enzymatically derived MOS also showed metal (Fe2+) chelating and anti-oxidant activities, wherein best anti-glycating agent was evaluated as MOS from PKC. PKC derived MOS showed highest cytotoxicity (74.19%) against human colon adenocarcinoma cell line (Caco-2). This study demonstrated the prebiotic potential of agro-waste derived MOS and possibility of their utilization as a functional food ingredient. Unlabelled Image • Aspergillus oryzae mannanase generated MOS from palm kernel cake and copra meal. • M3 (1.26 mg/mL) and M2 (0.68 mg/mL) were purified from guar gum hydrolysate. • Oligosaccharides were characterized using FTIR and 1H &13C NMR. • Anti-oxidant, anti-glycation and anti-cancerous activities of MOS were demonstrated. [ABSTRACT FROM AUTHOR]

Copyright of Revista Española de Quimioterapia is the property of Sociedad Espanola de Quimioterapia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The skin resembles an attractive target for vaccination due to its accessibility and abundance of resident immune cells. Cells like γδ T cells and mast cells (MCs) are part of the first line of defence against exogenous threats. Despite being important mediators for eliciting TH2 immune responses after epithelial stress, γδ T cell and MC functions still remain to be completely understood. Here, we aimed to characterize their roles in shaping adaptive immune responses after laser-mediated epicutaneous immunization (EPI). γδ T cell knock out, MC-depleted, and wildtype control mice were immunized with mannan-conjugated grass pollen allergen Phl p 5 (P5-MN) by laser-mediated EPI. After 2–3 immunizations, cytokine expression, T helper polarization, and antigen-specific IgG1/IgE levels were analysed. Furthermore, the local cytokine/chemokine milieu after laser microporation was determined. The majority of inflammatory chemokines and cytokines induced by laser treatment were not affected by the presence of γδ T cells or MCs. However, RANTES was elevated in γδ T cell knock out mice and GROα, TSLP, and IL-33 were significantly decreased after MC depletion. The absence of γδ T cells or depletion of MCs had no substantial effect on adaptive immune responses after laser-mediated EPI, except for slightly reduced IgG1 and effector T cell levels in MC-depleted mice. γδ T cells did not play a pivotal role in shaping the humoral and cellular adaptive immune response after laser-mediated EPI. MC depletion decreased the numbers of effector T cells, indicating a potential role of MCs in the activation and maturation of T cells after EPI. [ABSTRACT FROM AUTHOR]

• EPS production from Weissella confusa MD1 is optimized, purified and characterized. • This is the first reports of Mannan EPS production from Weissella confusa. • The average molecular weight of EPS was 2.909 KDa. • The EPS was thermally stable and showed degradation temperature around 267.74 °C. • The EPS showed promising antioxidant activity and antibiofilm activity. The present study focused on production, optimization and characterization of exopolysaccharide (EPS) from Weissella confusa MD1. The purified EPS MD1 was also evaluated for in vitro biological activities. The maximum yield of EPS (10.07 ± 0.32 g/L) was obtained with optimized culture conditions of 35 °C at pH 6.5 for 36 h with 4% (w/v) galactose and 1% (w/v) ammonium nitrate supplemented in MRS broth. The crude EPS was purified with diethylamino ethanol-Sepharose Fast Flow column and Sephadex-G 75 column. The physicochemical and functional characterization of the EPS was done by high-performance thin-layer chromatography (HPTLC), high-performance gel permeation chromatography (HPGPC), fourier transform infrared (FTIR) spectroscopy, nuclear magnetic resonance (NMR), thermo gravimetric and differential scanning calorimetric (TG-DSC) analysis, x-ray diffraction (XRD) and scanning electron microscopy (SEM). HPGPC analysis showed molecular weight of purified EPS MD1 as 2.909 KDa. Monosaccharide analysis showed that EPS was a novel mannan with mannose as the only monomeric unit present, suggesting EPS to be a homopolysaccharide containing → 6) α-Man p (1 → linkages. Microstructure studies by SEM showed MD1 EPS has globular and porous structure. The purified EPS MD1 exhibited higher thermal stability having degradation temperature around 267.74 °C with melting enthalpy value (Δ H) of 337.7 J/g. The EPS showed promising antioxidant activities with excellent antibiofilm activity against Staphylococcus aureus , Listeria monocytogenes , Salmonella enterica and Salmonella typhi. These striking physicochemical characteristics features and bioactivities of EPS would serve as potential candidate in food processing industry to be used as food adjunct in foods with health benefits. This is the first study reporting a mannan homopolysaccharide from Weissella sp. to be structurally characterized. [ABSTRACT FROM AUTHOR]

The interactions between tannins and soluble and insoluble cell wall components are, in part, responsible for the low quantities of tannins found in wines compared with the quantities in grapes. The use of polysaccharides to compete with cell wall components could be an interesting approach for improving the chromatic and sensory characteristics of wines. The e ect of two commercial polysaccharides, pectin and mannan, on limiting tannin-cell wall interactions was studied in a model solution, measuring the concentration of tannins and polysaccharides remaining in solution after the di erent interactions by chromatography. The treatment was also tested in a small-scale vinification. Soluble polysaccharides were added to the must and the wines were evaluated at the end of alcoholic fermentation and after six months in the bottle. In the model solution, the commercial polysaccharides formed soluble complexes with the tannins and limited the interactions with cell wall components, with some di erences between skin and seed tannins. In the case of the wines, the treatments resulted in wines with a higher color intensity and phenolic content. Sensory analysis resulted in higher scores for the wines with added polysaccharides, since the complexation of tannins with the polysaccharides increased the roundness and body of the resulting wines. [ABSTRACT FROM AUTHOR]

Apart from its role as a digestive and absorptive organ, the gastrointestinal (GI) tract is a vital immune organ that encompasses roughly 70 % of the total immune cells of the body. As such, the physical, chemical and nutrient composition of the diet influences overall GI function, effectively as an immune organ. With the improvement in feed technology, agro-industrial co-products that are high in fibre have been widely used as a feed ingredient in the diets of pigs and poultry. Arabinoxylan (AX) and mannan are the most abundant hemicellulosic polysaccharides present in cereal grain and co-product ingredients used in the livestock industry. When monogastric animals consume diets containing high amounts of AX and mannans, stimulation of GI immune cells may occur. This involves the activation of several cellular and molecular pathways of the immune system and requires a considerable amount of energy and nutrients to be expended by the animal, which may ultimately influence overall health and growth performance of animals. Therefore, a better understanding of the role of AX and mannan in immune modulation will be helpful in modulating untoward GI immune responses, thereby minimising nutrient and energy expenditure toward this effort. This review will summarise pertinent research on the role of oligosaccharides and polysaccharides containing AX and mannans in immune modulation in order to preserve gut integrity. [ABSTRACT FROM AUTHOR]

Mannoprotein (mannan) extracted from the cell wall of Saccharomyces cerevisiae is a bioactive glycoprotein that has many biological effects. In the present work, the preparation of gelatinbased blend films containing different concentrations of mannan (0, 0.5, 1.0, and 1.5%) was investigated. The effects of mannan on moisture content, water solubility, water vapor permeability (WVP), mechanical, optical, and thermal features of films were investigated. Moreover, the antifungal and aflatoxin B1 (AFB1) binding traits of the developed films were also studied. The results indicated that the addition of mannan significantly increased the water solubility, elongation at break, and WVP, whereas glass transition temperature and elastic modulus drastically decreased in the blend films. Furthermore, an obvious interaction between mannan and gelatin was concluded from Fourier-Transform Infrared (FTIR) spectra. Scanning electron microscopy assessment of the films revealed a heterogeneous structure. According to antifungal assay, the gelatin-mannan blend films had antifungal effect against Aspergillus flavus in disc diffusion evaluation. Similarly, these active films could reduce AFB1 content of pistachio extract agar. [ABSTRACT FROM AUTHOR]

Candida albicans is a leading cause of invasive candidiasis. C. albicans NBRC 1385 releases a mannoprotein beta-glucan complex (CAWS) into C-limiting medium. CAWS induced various biological activities, such as cytokine synthesis by leukocytes, platelet aggregation, acute lethal toxicity, induction of coronary arteritis, and the enhancement of side effects of indomethacin. CAWS is a ligand for dectin-2 and functions as pathogen associated molecular patterns (PAMPS). CAWS induced coronary arteritis (CAWS vasculitis) resembling that seen in patients with Kawasaki disease (KD). CAWS vasculitis has been applied to clarify the molecular mechanisms and to develop therapeutic strategies for KD. Detoxification of PAMPS gradually proceeds in the host by binding proteins, neutrophils, and macrophages and is the limiting factor to defme the time course of acute as well as chronic inflammatory diseases. To clarify the detoxification of CAWS by oxidative stress, CAWS were treated with NaOH, H2SO4, or sodium hypochlorite and tested for the modification of the structure, acute lethal toxicity and CAWS vasculitis. It was found that the structure and function of CAWS was stable with 1.0 N NaOH, 1.0 N H2SO4, or 2% sodium hypochlorite treatment. These data strongly suggest that the structure of CAWS is essential to induce pathological activities of Candida, and are resistant to oxidative detoxification stress in the host. [ABSTRACT FROM AUTHOR]

[Display omitted] • The complexation of SLP with mannan was predicted by molecular docking and molecular dynamics simulation. • The quenching mechanism of mannan to SLP belonged to static with high-affinity. • The interaction between mannan and SLP was mainly driven by Hydrophobic forces. • Mannan induced conformational changes of SLP structure. • The SLP-mannan complex was stable and compact. Tibet kefir grain (TKG) formation is mainly dependent on the aggregation of lactobacillus and yeasts. The interaction of surface layer protein (SLP) and yeast mannan plays an important role in mediating the co-aggregation of Lactobacillus kefiri with Saccharomyces cerevisiae. The interaction mechanism of the two was researched through multispectral spectroscopy, morphology observation and silico approaches. Fluorescence spectra data revealed that mannan was bound to SLP through a spontaneous binding process. The particle size of the binding complex increased as the mannan concentration increased. Synchronous fluorescence spectroscopy and circular dichroism (CD) spectra showed the conformational and microenvironment alteration of SLP treated with mannan. Molecular docking results indicated that hydrophobic interactions played major roles in the formation of SLP-mannan complexes. These findings provide a deeper insight into the interactions of protein and polysaccharide, and this knowledge is valuable in the application of SLP and mannan in co-fermentation systems. [ABSTRACT FROM AUTHOR]

[Display omitted] • Hexametaphosphate allows separation of proteins from extracellular polysaccharides. • Calcium ions effectively remove excess hexametaphosphate from protein solutions. • The purification method does not alter the secondary structure of β-lactoglobulin. • Yeast-based expression of β-lactoglobulin resulted in an extended N-terminus. • The extended N-terminus did not alter the structure of recombinant β-lactoglobulin. Proteins produced through precision fermentation are often purified through chromatographic methods. Faster and more cost-effective purification methods are desired for food application. Here, we present a simple method for purification of protein produced from yeast, using β-lactoglobulin secreted from Pichia pastoris as an example. The food-grade salt hexametaphosphate (HMP) was used to precipitate the protein at acidic pH, while the impurities (extracellular polysaccharides; mainly mannan) remained soluble. After re-solubilization of the protein-HMP complex by neutralization, excess HMP was selectively precipitated using calcium chloride. The protein content of the crude sample increased from 26 to 72 wt% (comparable to purification with anion exchange chromatography), containing only residual extracellular polysaccharides (9 wt%) and HMP (1 wt%). The established method had no significant impact on the structural and functional properties (i.e., ability to form emulsions) of the protein. The presented method shows potential for cost-effective purification of recombinant proteins produced through yeast-based expression systems. [ABSTRACT FROM AUTHOR]

A growing demand in novel food products for well-being and preventative medicine has attracted global attention on nutraceutical prebiotics. Various plant agro-processes produce large amounts of residual biomass considered “wastes”, which can potentially be used to produce nutraceutical prebiotics, such as manno-oligosaccharides (MOS). MOS can be produced from the degradation of mannan. Mannan has a main backbone consisting of β-1,4-linked mannose residues (which may be interspersed by glucose residues) with galactose substituents. Endo-β-1,4-mannanases cleave the mannan backbone at cleavage sites determined by the substitution pattern and thus give rise to different MOS products. These MOS products serve as prebiotics to stimulate various types of intestinal bacteria and cause them to produce fermentation products in different parts of the gastrointestinal tract which benefit the host. This article reviews recent advances in understanding the exploitation of plant residual biomass via the enzymatic production and characterization of MOS, and the influence of MOS on beneficial gut microbiota and their biological effects (i.e., immune modulation and lipidemic effects) as observed on human and animal health.

Among the several human fungal pathogens, Candida genus represents one of the most implicated in the clinical scenario. There exist several distinctive features that govern the establishment of Candida infections in addition to their capacity to adapt to multiple stress conditions inside humans which also include evasion of host immune responses. The complex fungal cell wall of the prevalent pathogen, Candida albicans, is one of the main targets of antifungal drugs and recognized by host immune cells. The wall consists of tiered arrangement of an outer thin but dense covering of mannan and inner buried layers of β-glucan and chitin. However, the pathogenic fungi adopt strategies to evade immune recognition by masking these molecules. This capacity to camouflage the immunogenic polysaccharide β-glucan from the host is a key virulence factor of C. albicans. The present review is an attempt to collate various underlying factors and mechanisms involved in Candida β-glucan masking from the available pool of knowledge and provide a comprehensive understanding. This will further improve therapeutic approaches to candidiasis by identifying new antifungal targets that blocks fungal immune evasion.

Monoclonal antibody EBCA-1 is used in the sandwich immune assay for the detection of circulating Candida mannan in blood sera samples for the diagnosis of invasive candidiasis. To reinvestigate carbohydrate specificity of EBCA-1, a panel of biotinylated oligosaccharides structurally related to distinct fragments of Candida mannan were loaded onto a streptavidin-coated plate to form a glycoarray. Its use demonstrated that EBCA-1 recognizes the trisaccharide β-Man-(1→2)-α-Man-(1→2)-α-Man and not homo-α-(1→2)-linked pentamannoside, as was reported previously.

A trial was conducted to investigate the effects of dietary mannan level and β-mannanase supplementation on egg production performance, nutrient retention and blood metabolites of laying hens. Two hundred and forty Hy-Line Brown layers (52 wk-old) were randomly allotted to 6 treatments on the basis of laying performance. Each treatment had 8 replicates with 5 birds (40 birds per treatment). Laying hens were fed low or high mannan diets containing 0, 0.4 or 0.8 g β-mannanase/kg diet in a 2×3 factorial arrangement during 56 d feeding period. Laying hens fed diets supplemented with high β-mannanase level had greater (P

The exogenous enzymes are less consistent in their effects as their beneficial effects depend upon the types and level non-starch polysaccharides (NSP) present in the diets. Therefore, exogenous enzymes should be selected on the basis of types and amount of the NSP in the pig diets. The objectives of the present experiments were to investigate the effects of dietary level of mannan and β-mannanase supplementation on growth performance, apparent total tract digestibility (ATTD) of energy and nutrients, and blood metabolites of growing pigs. In Exp. 1, 96 barrows were randomly allotted to four treatments on the basis of BW. There were four replicates in each treatment with six pigs per replicate. The dietary treatments were a corn–soybean meal (SBM)-based control diet and three other diets consisted of the control diet supplemented with 400, 800 or 1600 U of β-mannanase/kg diet. The final BW, average daily gain (ADG) and blood glucose increased (linear, P0.05) on the concentrations of blood total cholesterol, triacylglycerides and blood urea nitrogen. These results indicate that supplementation of β-mannanase to low- or high-mannan diets have potential to improve the performance of growing pigs. In addition, palm kernel meal may partially replace corn and SBM without reducing pig performance if β-mannanase is added to diet.

ABSTRACT Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus cause life-threatening candidiasis, cryptococcosis, and aspergillosis, resulting in several hundred thousand deaths annually. The patients at the greatest risk of developing these life-threatening invasive fungal infections have weakened immune systems. The vulnerable population is increasing due to rising numbers of immunocompromised individuals as a result of HIV infection or immunosuppressed individuals receiving anticancer therapies and/or stem cell or organ transplants. While patients are treated with antifungals such as amphotericin B, all antifungals have serious limitations due to lack of sufficient fungicidal effect and/or host toxicity. Even with treatment, 1-year survival rates are low. We explored methods of increasing drug effectiveness by designing fungicide-loaded liposomes specifically targeted to fungal cells. Most pathogenic fungi are encased in cell walls and exopolysaccharide matrices rich in mannans. Dectin-2 is a mammalian innate immune membrane receptor that binds as a dimer to mannans and signals fungal infection. We coated amphotericin-loaded liposomes with monomers of Dectin-2’s mannan-binding domain, sDectin-2. sDectin monomers were free to float in the lipid membrane and form dimers that bind mannan substrates. sDectin-2-coated liposomes bound orders of magnitude more efficiently to the extracellular matrices of several developmental stages of C. albicans, C. neoformans, and A. fumigatus than untargeted control liposomes. Dectin-2-coated amphotericin B-loaded liposomes reduced the growth and viability of all three species more than an order of magnitude more efficiently than untargeted control liposomes and dramatically decreased the effective dose. Future efforts focus on examining pan-antifungal targeted liposomal drugs in animal models of fungal diseases. IMPORTANCE Invasive fungal diseases caused by Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus have mortality rates ranging from 10 to 95%. Individual patient costs may exceed $100,000 in the United States. All antifungals in current use have serious limitations due to host toxicity and/or insufficient fungal cell killing that results in recurrent infections. Few new antifungal drugs have been introduced in the last 2 decades. Hence, there is a critical need for improved antifungal therapeutics. By targeting antifungal-loaded liposomes to α-mannans in the extracellular matrices secreted by these fungi, we dramatically reduced the effective dose of drug. Dectin-2-coated liposomes loaded with amphotericin B bound 50- to 150-fold more strongly to C. albicans, C. neoformans, and A. fumigatus than untargeted liposomes and killed these fungi more than an order of magnitude more efficiently. Targeting drug-loaded liposomes specifically to fungal cells has the potential to greatly enhance the efficacy of most antifungal drugs.