• Food Science and Biotechnology
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• v.18 no.3
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• pp.667-671
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• 2009

Extract of water-soluble polysaccharide (CFWx), showing inhibiting activity on ${\alpha}-glucosidase$, was prepared from the fruiting bodies of Cordyceps militaris by hot-water extraction, and ethanol precipitation. Chemical characteristics of CFWx were as follows: carbohydrate content 30% including 16% of uronic acid; 51% protein content; monosaccharide composition, Man:Glu:Gal (30:43:27); molecular weight $3-5{\times}10^4$. CFWx was further purified by ion-exchange, gel-permeation, and affinity chromatography and $CFWx-AH-{\alpha}$ fraction was isolated. Fundamental structure of $CFWx-AH-{\alpha}$ was deduced as ${\alpha}-(1{\to}4$)-D-glucan with ${\alpha}-(1{\to}3$)- and/or ${\alpha}-(1{\to}6$)-D-glycosidic side chains based on methylation analysis.

• Journal of Life Science
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• v.21 no.11
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• pp.1579-1585
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• 2011

Natural anti-diabetic semipurified ethyl acetate fraction was isolated from the submerged-liquid culture of Agaricus blaze Murill (AB) in a medium containing soybean flakes. Hot-water extract of AB (HEAB) was prepared by extraction at $121^{\circ}C$ for 60 min, followed by filtering through a filter presser filled with diatomate. The ${\beta}$-glucan-free HEAB, which was a supernatant fraction from HEAB by precipitation in an 80% ethanol solution, was fractionated into hexane, chloroform, ethyl acetate, and butanol fractions. The inhibition of the ${\alpha}$-glucosidase activity by fractions was 59.0, 17.0, 61.6, and 37.9%, respectively, suggesting that ethyl acetate fraction was the most active. A subfraction having a strong ${\alpha}$-glucosidase inhibitory activity (80.4%) was isolated from the ethyl acetate fraction. This subfraction contained isoflavones (genistin and daidzin) and their conjugates with sugars as potent inhibiters of ${\alpha}$-glucosidase activity. These results suggest that the ethyl acetate fraction or HEAB containing isoflavones and their sugars conjugates could be useful sources for controlling blood sugar levels in humans.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.69-77
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• 2019

Screening microorganisms that can produce glucan hydrolases for industrial, environmental, and biomedical applications is important. Herein, we describe a novel approach to perform glucan hydrolase screening-based on analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) spectra-which involves degradation of the oligo- and polysaccharides. As a proof-of-concept study, glucan hydrolases that could break down glucans made of several glucose units were used to demonstrate the MALDI-MS-based enzyme assay. First, the enzyme activities of ${\alpha}$-amylase and cellulase on a mixture of glucan oligosaccharides were successfully discriminated, where changes of the MALDI-MS profiles directly reflected the glucan hydrolase activities. Next, we validated that this MALDI-MS-based enzyme assay could be applied to glucan polysaccharides (i.e., pullulan, lichenan, and schizophyllan). Finally, the bacterial glucan hydrolase activities were screened on 96-well plate-based platforms, using cell lysates or samples of secreted enzyme. Our results demonstrated that the MALDI-MS-based enzyme assay system would be useful for investigating bacterial glucoside hydrolases in a high-throughput manner.

• Mycobiology
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• v.42 no.2
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• pp.167-173
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• 2014

A ${\beta}$-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble ${\beta}$-1,3-glucan (${\beta}$-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a ${\beta}$-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal ${\beta}$-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal ${\beta}$-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa ${\beta}$-glucan synthase was estimated to include catalytically insignificant transmembrane ${\alpha}$-helices and loops. Sequence analysis of 101 fungal ${\beta}$-glucan synthases, obtained from public databases, revealed that the ${\beta}$-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of ${\beta}$-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa ${\beta}$-glucan synthase in this study belonged to Type II family, meaning Type I ${\beta}$-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble ${\beta}$-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa ${\beta}$-glucan synthase will provide better explanations.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.438-445
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• 2010

The immunomodulatory effects of Aureohasidium pullulans SM-2001 exopolymers containing $\beta$-1,3/1,6-glucan were evaluated in cyclophosphamide (CPA)-treated mice. To induce immunosuppression, 150 and 110 mg/kg of CPA were intraperitoneally injected 3 days and 1 day, respectively, before beginning administration of the test material. Exopolymers were delivered subcutaneously or orally, four times, in a volume of 10 ml/kg at 12-h intervals beginning 24 h after the second CPA treatment. Changes in thymus and spleen weights, splenic amounts of tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-$1{\beta}$, and IL-10, and numbers of CD3+, CD4+, CD8+, and TNF-$\alpha+$ thymus and spleen cells were monitored in CPA-treated mice. As a result of CPA treatment, dramatic decreases in the number of CD3+, CD4+, CD8+, and TNF-$\alpha+$ cells were detected in the thymus and spleen, along with decreases in thymus and spleen weights. In addition, splenic TNF-$\alpha$, IL-$1{\beta}$, and IL-10 contents were also decreased on observation with flow cytometry. However, oral and subcutaneous treatments with exopolymers effectively reduced the immunosuppressive changes induced by CPA. Therefore, it is concluded that exopolymers of A. pullulans SM-2001 can effectively prevent immunosuppression through, at least partially, the recruitment of T cells and TNF-$\alpha+$ cells or enhancement of their activity, and can provide an effective component of prevention or treatment regimens for immunosuppression related to cancer, sepsis, and high-dose chemotherapy or radiotherapy.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.12
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• pp.1753-1759
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• 2008

Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2007.11a
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• pp.27-31
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• 2007

This study relates to low and medium molecular weight isoflavone-${\beta}$-D-glucan produced by submerged liquid culture of Agaricus blazei, a method of producing the isoflavone-B-D-glucan using autolysis enzyme of Agaricus blazei mycelia, and use of the isoflavone-B-D-glucan for anti-cancer and immunoenhancing effect. In acordance with one aspect of the present study, it deals with a method of producing isoflavone-${\beta}$-D-glucan, which comprises the followings; 1) culturing and separating mushroom mycelia, 2) producing low-medium molecular weight isoflavone-${\beta}$-D-glucan from high molecular weight one. The cytotoxicity on human cnacer cell line (Caco-2, MCF-7), the expression of Cyclin D, Bcl-2, Bax protein, p21 protein, p53 protein in MCF-7 cells assessed by SDS-PAGE and immunoblotting, and other immuno related factors such as TNF-${\alpha}$ and IL-1B activities were examined. Structural identification of isoflavone-${\beta}$-D-glucan which showed cytotoxicity against cancer cell and immunoenhancing effects was carried by separation with DEAE-cellulose column chromatography, TLC, HPLC, IR, NMR. Clinical test for the cancer patients (n=119) for 6 month was carried out, and immunoenhancing factors (NK. cell number, ratio of T4/T8) were checked. We concluded the identified isoflavone-${\beta}$-D-glucan has immuno enhancing effects and could be useful for cancer chemoprevention.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.2
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• pp.237-244
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• 2008

The object of this trial was to investigate the effect of dietary ${\beta}$-1,3/1,6-glucan supplementation on the performance and immunological response of broiler chickens. Two hundred and forty 1-day old male broilers ($39{\pm}1g$) were separated into six treatments which were given six different feeds containing 0 (control), 25, 50, 75, 100 and 125 mg/kg dietary ${\beta}$-1,3/1,6-glucan supplementation. On days 21 and 42, body weight gain, feed consumption and feed conversation rate were recorded as measures of growth performance. The levels of key cytokines in the immuno-regulating pathway: interleukin-1 (IL-1), interleukin-2 (IL-2), interferon $\gamma$(IFN-$\gamma$, tumor necrosis factor $\alpha$(TNF-$\alpha$, and the concentrations of signal molecules: peripheral blood plasma globulin, serum Immunoglobulin G (IgG) and intestinal secretary Immunoglobulin A (sIgA), were measured as indices of the immune response to determine suitable levels of dietary ${\beta}$-1,3/1,6-glucan supplementation. The results indicated that performance was elevated quadratically with dietary ${\beta}$-1,3/1,6-glucan supplementation. Maximal growth performance and an enhanced immunological response were obtained at a supplemented level of 50 mg/kg.

• Korean Journal of Food Science and Technology
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• v.25 no.1
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• pp.15-21
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• 1993

Crude ${\beta}-glucan$ extracted from Barley was purified by stepwise enzyme treatment (Thermostable ${\alpha}-amylase$, amyloglucosidase, protease). The Intrinsic Viscosity $[{\eta}]$ of the purified ${\beta}-glucan$ was determined by Cannon Fenske Capillary Viscometer (size 50, Cannon Instruments, State, College pa.) at different pH (2, 4, 7, 9, 11) and various salt concentration (0.01 M, 0.03 M, 0.05 M, 0.07 M, 0.1 M and 0.2 M). The $[{\eta}]$ of purified ${\beta}-glucan$ was ranged from $0.997{\sim}2.290\;dl/g$. The $[{\eta}]$ of purified ${\beta}-glucan$ at both alkali, acid condition were lower than those at pH 7. However, the alkali condition of puified ${\beta}-glucan$ solution showed less $[{\eta}]$ than the acid condition of this solution. From 0 M to 0.2 M salt concentration, the $[{\eta}]$ of purified ${\beta}-glucan$ solution was decreased to 0.03 M then increased to 0.05 M NaCl and remained constant to 0.2 M NaCl. The chain stiffness parameter of purified ${\beta}-glucan$ was not affected by temperature from $15^{\circ}C$ to $65^{\circ}C$. The shear rates of various ${\beta}-glucan$ conditions were determined by Bohlin Rheometer (Lund, Sweden). The ${\beta}-glucan$ concentration of 1.0 g/dl and 2.0 g/dl behaved as Newtonian fluid. However, above the concentration of 3.0 g/dl ${\beta}-glucan$ solution, it showed thixotropic and psedoplastic characteristics. Barley ${\beta}-glucan$ appears a damping at 0.5 frequency for the 4.0 g/dl solution. Below 0.5 frequency, it appears a viscous behavior property and above 0.5 frequency, it appears a elastic behavior property.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.267-271
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• 2000

Abstract Two glucans (PONGa and PONGb) differing in their anomeric and glycosidic linkage structures were isolated from the water-insoluble materials (PON) of Pleurotus ostreatus basidiocarps, which activated the complement system and were almost soley composed of D-glucose. The isolatIon was achieved by repeated precipitations with ethanol and adsorption on concanavalin A (Con A) of paN suspension in thymol/NaCL Based on methylation analysis. IR, GLC-MS, $^1H,{\;}and{\;}^{13}C-NMR$ spectroscopies, PONGa was found to be a branched a-glucan composed of ${\alpha}-linked$ D-glucopyranose residues and ${\alpha}-linked$ units with 6-branching points, whereas PONGb was a linear ${\beta}-1,3-glucan$ composed mainly of ${\beta}-1,3-linked$ D-glucopyranose residues. The PONGb particles reacted more potently than the PONGa particles as C3 activator in alternative complement hemolysis and crossed-immunoelectrophoresis using anti-human C3, thereby suggesting that the complement activating components of PON were ${\beta}-(13)-glucans rather$ than ${\alpha}-glucan$ components.onents.