• Applied Biological Chemistry
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• v.41 no.1
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• pp.60-66
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• 1998

During the screening for anti-complementary activity from 10 kinds of edible mushrooms, an alkali extract of Agaricus bisporus showed the highest activity through the complement fixation test. The crude anti-complementary material(AB-0) from Agaricus bisporus was obtained by the alkali extraction using 1 N NaOH containing 5% urea$(65^{\circ}C)$, followed by methanol reflux, dialysis and lyophilization. The fraction AB-O showed potent anit-complementary and anti-tumor activity against sarcoma-180 injected mice. The fraction AB-0 was divided into 5 fractions(AB-20, AB-40, AB-60, AB-80, AB-A) by gradual acetone precipitation. Among them fraction AB-20 having the highest activity and yield was found to contain 39% carbohydrate and 46% protein. The anti-complementary protein-bound polysaccharide AB-20 consisted of glucose, arabinose, xylose, galactose and mannose in a molar ratio of 6.49 : 1.98 : 1.24 : 1.00 : 0.71, respectively and its main component amino acids were alanine(20.59%), isoleucine(16.85%), glutamine+glutamic acid(14.12%) and leucine (13.83%). The anti-complementary activity of AB-20 was decreased greatly by periodate oxidation, but decreased slightly by pronase digestion. This indicates that polysaccharide moiety is corelated with the anti-complementary activity and that protein is also involved in the activity.

• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2113-2120
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• 2018

Cross-reactive material 197 ($CRM_{197}$) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. $CRM_{197}$ has been used as a carrier protein for poorly immunogenic polysaccharide antigens to improve immune responses. In this study, to develop a sandwich ELISA that can detect $CRM_{197}$ and $CRM_{197}$ conjugate vaccines, we generated a human anti-$CRM_{197}$ monoclonal antibody (mAb) 3F9 using a phage-displayed human synthetic Fab library and produced mouse anti-$CRM_{197}$ polyclonal antibody. The affinity ($K_D$) of 3F9 for $CRM_{197}$ was 3.55 nM, based on Bio-Layer interferometry, and it bound specifically to the B fragment of $CRM_{197}$. The sandwich ELISA was carried out using 3F9 as a capture antibody and the mouse polyclonal antibody as a detection antibody. The detection limit of the sandwich ELISA was <1 ng/ml $CRM_{197}$. In addition, the 3F9 antibody bound to the $CRM_{197}$-polysaccharide conjugates tested in a dose-dependent manner. This ELISA system will be useful for the quantification and characterization of $CRM_{197}$ and $CRM_{197}$ conjugate vaccines. To our knowledge, this study is the first to generate a human monoclonal antibody against $CRM_{197}$ and to develop a sandwich ELISA for $CRM_{197}$ conjugate vaccines.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.450-456
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• 1999

A high molecular-weight water-soluble fraction(PO) obtained by the ethanol precipitation of 0.1 N NaOH extracts of the mushroom Pleurotus ostreatus showed 82% anti-complementary activity for complement consumption hemolysis. The PO consisted of 42% carbohydrate (w/w), 50% protein (w/w), and 3% uronic acid (w/w). Fifty-eight percent of the anti-complementary activity decreased by periodate oxidation and 22% by protease digestion, suggesting that the sugar and protein moieties are essential for this activity. Two polysaccharide fractions, PO-IIIa-1 and PO-IIIa-2, with anti-complementary activity were isolated from the PO using DEAE-Sepharose FF followed by Sephadex G-75 and Sepharose CL-6B gel permeation chromatographies. The PO-IIIa-2 was found by HPLC to be nearly homogeneous, with the molecular mass of 531 kDa, and showed 96% $ITCH_{50}$ (inhibition against the total complement hemolysis of deionized water as the control) at a concentration of 1 mg/ml. This fraction contained galactose, mannose, fucose, and glucose with molar ratios of 1.75:1:0.65 and 0.59, respectively. The majority of galactose and mannose units in the PO-IIIa-2 were composed of TGalp1 ->, ->6Galp1->, ->2,6Galp1->, and ->Manp1->. The PO-IIIa-1 (molecular mass of 2000 kDa), exhibiting higher activity than the PO-IIIa-2, was further purified into two fractions, unbound proteoglycan (PO-IIIa-1A) and bound glucomannan (PO-IIIa-lB), by affinity chromatography using ConA-Sepharose CL-4B. The anti-complementary activity of each affinity purified fraction decreased as compared to that of the native PO-IIIa-1 fraction, indicating that the formation of complex between both polysaccharide fractions was necessary for full anti-complementary activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.6
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• pp.1183-1188
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• 1998

This study was carried out to elucidate the immunomodulating activity of protein bound polysac charide extracted from Ganoderma lucidum(PSG). Macrophage drived reactive radicals were known as an effector for antimicrobial and anticancer functions. The promising immune response molecule, reactive oxygen intermediates(ROIs), was determined in TIB 71 cells with PSG at various experimental conditions. Treatment with 0.5mg PSG significantly increased the production of ROIs, superoxide anion as well as hydrogen peroxide, from TIB 71 cells(p<0.001). Under the same concentration, con siderable results were obtained from 24 hour cultivation with 106 cells at 5% CO2 incubator. The cells were trigged with PMA(5.3 M) after primed BCG(100 M) or IFN (100U) alone could not induce the production of ROIs, but it had a significant potentiating effect on ROIs secretion when the cells were treated with PSG.

• Archives of Pharmacal Research
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• v.2 no.2
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• pp.145-151
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• 1979

The carpophores of three Korean mushrooms, Coriolus versicolor, Pleurotus ostreatus and Lentinus edodes were respectively extracted with hot water and the extracts were dialyzed through Visking tube. They were found to exert an antitumor activity against sarcoma-180 implanted in mice. Especially, the inhibition ratio of the extract of Coriolus versicolor (100 mg/ Kg.i.p) was almost 100% . But all the extracts did not affect the growth of leukemia L517Y cells in vitro. Therefore these facts indicate that the extracts appear to stimulate cell-mediated immunity.

• The Korean Journal of Mycology
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• v.10 no.4
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• pp.213-216
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• 1982

To find antitumor components with low toxicity in natural products of Korean Basidiomycete, the carpophores of Flammulina velutipes (Fr.) Sing. were extracted with hot water for eight hours. The extract was purified by dialyzing through Visking tube and a protein-bound polysaccharide fraction was obtained as pale brownish amorphous powder after it was freeze-dried. The fraction was examined for antitumor activity against sarcoma 180 implanted subcutaneously in the left groins of I.C.R. mice. The inhibition ratio of this fraction of against the tumor was 62.3% at the dose of 10 mg/kg/day for the period of ten days. The tumors in three of the ten treated mice were completely regressed.

• The Korean Journal of Mycology
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• v.13 no.3
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• pp.131-139
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• 1985

To find antitumor constituents in the shake cultured mycelia of Agaricus campestris, the mycelia were extracted with hot water. Purification of the extract was carried out by ethanol precipitation and by ion exchange chromatography using DEAE-Sephadex A-50. Each fraction obtained during the purification procedure was examined for antitumor activity against sarcoma 180 in ICR mice. The antitumor fraction C was isolated. It showed 56.1% inhibition ratio at a dose of 20 mg/kg/day and consisted of a polysaccharide moiety (45%) and a protein moiety (18%). The polysaccharide was analyzed by G.L.C. and found to contain mannose (42.0%), glucose (25.5%), xylose (16.6%), fucose and galactose. The protein moiety was composed of 17 amino acids. The antitumor fraction A showed immunopotentiating activity by accumulating peritoneal macrophages and by increasing plaque-forming cells in mice.

• Food Science and Biotechnology
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• v.18 no.6
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• pp.1386-1391
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• 2009

Crude polysaccharides from rapeseed meal (PRM) were extracted with 0.3% NaOH aqueous solution, followed by further purifications and 2 fractions, namely PRM1 and PRM2, were separated with a DEAE-cellulose DE-52 column. Their primary compositions were analysed and antioxidant activity was determined, including scavenging activity toward superoxide anion radicals, hydroxyl radicals, and nitric oxide radicals, reducing power, and inhibitory effects against the microsomal lipid peroxidation, compared to that of L-ascorbic acid. The results indicated that PRM1 and PRM2 exhibited not only good reducing power and inhibitory effects on the microsomal lipid peroxidation, but also strong scavenging activity toward superoxide anion radicals, nitric oxide radicals, and hydroxyl radicals. In addition, positive correlations were also observed between the superoxide anion radical scavenging activity and the protein contents of the polysaccharides, and the reducing power and the sulfate contents. These findings thus clearly suggest the polysaccharides possess direct and potent antioxidant activity.

• The Korean Journal of Mycology
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• v.22 no.2
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• pp.184-189
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• 1994

The possibility of solid-substrate fermentation of Coriolus versicolor for the production of protein-bound polysaccharides(PBP) was studied. Zeolite and orchid-pot soil were used as solid materials for the culture because of the desirable physical properties. Glucose, sucrose and starch showed to be good carbon sources for the production of PBP by the solid-substrate fermantation of C. versicolor. Among the nitrogen sources, bactosoyton and peptone were very effective for the PBP production. The optimum pH for solid-substrate culture for the production of PBP was at the range of 5-6. The yields of PBP reached to 5-6 mg per 100 g solid-substrate.

• Food Science and Preservation
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• v.11 no.1
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• pp.47-52
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• 2004

This study was conducted to investigate the characteristics of ultrafiltration and spray drying process for crude protein bound polysaccharide(CPBP) isolated from Agaricus blasei Murill. In ultrafiltration process, the permeate flux increased with the increase of operating pressure and temperature. The permeate flux declined continuously while the fouling materials were accumulated on the membrane as the operation time increased. In comparing of raw CPBP and filtered CPBP, the viscosity of CPBP treated UF was decreased and $\Delta$E value of treated samples was increased. Thermal efficiencies of spray drying process were increased by increasing inlet temperature, feed rate and feed concentration.