• Journal of Aquaculture
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• v.21 no.4
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• pp.325-330
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• 2008

Two different Eelwardsiella tarda isolates, KFE and Edk-2, were obtained from Korea and Japan respectively. On the base of the previous results showing higher pathogenicity of E. tarda KFE compared to that of E. tarda Edk-2 isolate, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Concentration of specific antibody in the serum appeared to be much higher in loach immunized with FKC of E. tarda Edk-2 than those found in loach immunized with FKC of E. tarda KFE. Cross-reaction analysis using agglutination test with normal and antigen-absorbed antisera implied the differences epitopes in the antigens of these E. tarda isolates. For the comparison of bactericidal activity of the produced antibody with different antigens, absorption analysis was performed and confirmed the presence of critical epitopes in the FKC of E. tarda KFE strain. The prophylactic agent, polysaccharide-bound protein (PS-K) injected 1 week before the artificial infection with E. tarda KFE isolates decreased the cumulative mortality in loach and would be on effective method to prevent the occurrence of bacterial infection including E. tarda.

• Journal of fish pathology
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• v.10 no.1
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• pp.45-52
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• 1997

A protein-bound polysaccharide preparation, PS-K, isolated from Coriolus versicolor was evaluated for its ability to enhance resistance against Edwardsiella tarda, causal agent of classical edwardsiellosis in tilapia, Oreochromis niloticus. Fish treated with a dose rate of 0.1 mg PS-K $g^{-1}$ body weight were challenged by intraperitoneal injection of E. tarda with different concentrations. Against low burdens of injections($1.2{\times}10^7$ and $1.2{\times}10^8$ cfu/fish), PS-K treated fish did not show any mortality compared to 50% and 100% mortality of control groups, respectively. However there was no increase of resistance against challenge with high concentrations of E. tarda ($1.2{\times}10^9$ and $1.2{\times}10^{10}$cfu /fish) except few days delaying of death. Tilapia injected with PS-K one day after intraperitoneal inoculation of E tarda($1.2{\times}10^7$ cfu/fish) showed 100% survival rate compared to control group of 50% survival rate. The result indicates the potential of PS-K as a prophylactic agent in aquaculture. The increased antibody response in fish received PS-K one week prior to FKC administration suggested the influence of PS-K on the specific humoral immunity and increased resistance against bacterial infection.

• Korean Journal of Food Science and Technology
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• v.36 no.4
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• pp.586-593
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• 2004

Efficient extraction method of crude protein-bound polysaccharide (CPBP) from Agaricus blasei Murill was established. CPBP yields by ultrasonic and hot water extractions were 13.0 and 7.8%, respectively. Pressure extraction for 3 hr gave the highest ${\beta}-glucan$ content; no significant difference was observed between 2 and 3 hr extraction. Four volumes added ethanol gave the highest yields of CPBP and ${\beta}-glucan$ contents at 10.89 and 35.97%, respectively. Decomposition temperature of CPBP was $240-365^{\circ}C$, showing relatively good thermal stability. In SRB (sulforhodamine B) assay, CPBP treatment at $1,000\;{\mu}g/mL$ for 72 hr inhibited proliferations to A549, MCF-7, and AGS cancer cells by 43.9, 21.4, and 32.5%, respectively.

• The Korean Journal of Mycology
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• v.23 no.2 s.73
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• pp.121-128
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• 1995

The characteristics of protein-bound polysaccharides (PBP) which were isolated and purified from the sawdust mycelia of Agrocybe cylindracea were investigated. The yield of crude protein-bound polysaccharides (Fr.CB) extracted with boiling water and precipitated with 95% ethanol, was 0.74% based on the original sawdust mycelia. The Fr.CB was purified by the membrane filtration, ion exchange chromatography and gel filtration. The Fr.B fraction of which the molecular weight is over 300 KDa, was isolated from the Fr.CB using membrane filtration, and the yield was 38.6% based on the Fr.CB. This result indicates that high molecular protein-bound polysaccharides are the dominent components of the Fr.CB. Two fractions (Fr.B-1, Fr.B-2) were also isolated from the Fr.B using ion exchange chromatography, and the yields were 17.3% (Fr.B-1) and 10.3% (Fr. B-2), respectively. The Fr.B-1 was concentrated and gel-filtrated, and the single peak, thought to be nearly pure protein-bound polysaccharides, was obtained. The yield of final fraction $(Fr.B-1-{\beta})$ was 42.5% based on the Fr.B-1. The molecular weight of $Fr.B-1-{\beta}$ was nearly 710 KDa. The monosaccharides' composition of $Fr.B-1-{\beta}$ was analized by HPLC, and glucose was the dominent component, and fucose and galactose were also detected. The result of amino acid analysis was that glutamic acid and analysis were detected to a significant level, and cysteine was not detected.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.702-708
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• 1998

The effects of mushrooms such as Lentinus edodes and Pleurotus ostreatus on anti-cancer activity through in vivo and in vitro experiments were powders of protein-bound polysaccharides in mushrooms were solubilized in 0, 5, 25 mg/kg saline, respectively and were used in vitro experiments. The in vivo experiments were carried out as followed: i) anti-cancer activities on Leukemia $(L_{1210})$, Hepatlicus cancer $(H_{22})$ and Sarcoma180/(S180), and ii) the effect on immune system through changes in intestine weight and the number of hemolytic plague forming cells. Protein-bound polysaccharides of all showed anti-cancer activity on $L_{1210}$ and fruit body of Lentinus edodes 25 mg/kg treatment group showed the highest inhibition rate (86%). Pleurotus ostreatus mycelial in medium of cultivate 25 mg/kg treatment. Fruid body of Lentinus edodes 25 mg/kg treatment group showed the highest inhibition rate (86% and 71%, respectively) on $H_{22}$ among them. The inhibition rates of fruit body and mycelial of Lentinus edodes 25 mg/kg treatment groups on S180 were 33.9% and 30.9%, respectively. Each samples of 50, 100, 200, $400\;{\mu}g/{\mu}L$ on in vitro cell toxicity test did not show significantly different cell death rates at P<0.05. In immune test, weights of liver and spleen were increased according to increase in conc. but were not significantly different at P<0.05. The weights of thymus were heavy in fruit body and mycelial of Lentinus edodes treatment group but were not significantly different at P<0.05. Hemolytic plague forming cells with antibody formation capability were significantly high in fruit body and mycelial of Lentinus edodes treatment samples.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.141-141
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• 1993

말징버섯 Calvatia craniformis의 culture broth 40 liter를 여과하여 얻은 균사채를 열수 추출하여 진한 갈색 건조분말(Fr. A) 13.1g을 분리하였다. Fr. A에 대하여 DEAE-cellulose ion chromatography를 시행하여 중성분획인 흰색 분말 (Fr. B) 2.50g을, 산성분획인 갈색 분말 (Fr. C) 3.50g을 각각 분리하였다. Fr. B 500mg에 대하여 Sepharose CL-4B gel filtration chromatography를 시행하여 흰색분말 Fr. D(Calvatan) 350mg을 분리하였다. Calvatan 350mg에 대하여 Con A-Sepharose 4B affinity chromatography를 적용하여 미흡착 분획인 Fr. F($\alpha$-form)와 흡착 분획인 Fr. E ($\beta$-form)로 정제하였다. 항암작용의 기전을 구명하고자 마우스에 대하여 면역학적 실험을 시행하였다. macrophage의 활성에 대한 영향을 조사하였던 바, 투여군의 활성화된 macrophage에 의해 유리되는 superoxide anion의 양은 대조군에 비해 1.4배 증가하였고 Calvatan 투여군의 용혈반 형성세포(PFC)는 대조군에 비해 3.1배 증가하였다. 화학 분석에 의해, Calvatan은 다당체 87.2%, 단백질 1.8% 및 hexosamine 1.3%로 구성되어 있었다. 따라서 항암성 분획들은 protein-bound polysaccharide임을 알 수 있었다. 또한 각 분획의 다당체를 구성하고 있는 단당류는 glucose, galactose, mannose 및 xylose 였으며 단백질 부분은 16종의 아미노산으로 구성되어 있었다. IR 스펙트럼은 3300-3400 $cm^{－1}$에서 0-H stretching frequency, 2900 $cm^{－1}$ 에서 C-H stretching frequency, 1630 $cm^{－1}$ 에서 C-0 stretching frequency. 1000-1100 $cm^{－1}$에서 C-H 및 C-0 bending frequency를 나타내는 다당체의 전형적인 특성을 보여주었다.을 보여주었다.

• Microbiology and Biotechnology Letters
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• v.46 no.1
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• pp.9-17
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• 2018

The aims of this work were to characterize and determine bioactivities of crude exopolysaccharide (EPS) extract from Bacillus tequilensis PS21 isolated from milk kefir from Kampaeng Petch, Thailand. B. tequilensis PS21 produced 112.1 mg dried EPS/l from initial 80 g/l lactose in modified TSB media at 52 h, with EPS product yield of 8.9 mg EPS/g lactose and specific product yield of 0.3 mg EPS/mg biomass. The FTIR result confirmed EPS to be a protein-bound polysaccharide and SEM analysis showed the morphology to be a grainy appearance with an uneven surface, covered with pores. HPLC analysis determined EPS as a heteropolysaccharide consisting of five sugar units with the following molar ratios; xylose (17.65), glucose (2.54), ribose (1.83), rhamnose (1.23), and galactose (1). Chemical components of this EPS were predominantly carbohydrate at 697.8 mg/g EPS (65%), protein 361.4 mg/g EPS (34%), and nucleic acid 12.5 mg/g EPS (1%). The EPS demonstrated antioxidant activities at 57.5% DPPH scavenging activity, $37.2{\mu}M\;Fe(II)/mg$ EPS and $34.9{\mu}M\;TEAC\;{\mu}M/mg$ EPS using DPPH, FRAP and ABTS assays, respectively. EPS also exhibited anti-tyrosinase activity at 34.9% inhibition. This work represents the first finding of EPS produced by Bacillus sp. from Thai milk kefir which shows potential applications in the production of antioxidant functional foods and whitening cosmetics. However, optimization of EPS production for industrial exploitation requires further study to ascertain the economic potential.

• The Korean Journal of Mycology
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• v.18 no.2
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• pp.58-69
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• 1990

ABSTRACT: In order to find physiologically active components from higher fungi, hot-water soluble components were extracted from the basidiocarps of Ganoderma lucidum. The extract was purified and separated by DEAE cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions were designated CR, IN, IA, GL and GH. Fraction GL showed the highest antitumor activity among the fractions and its molecular weight was found to be 47 KD. The tumor inhibition ratio of Fr. GL was 81 % at the dose of peritoneal administration of 20 mg/kg/day for 10 days in mice. Chemical analysis of this fraction showed 82% polysaccharide, 8% protein and 0.9% hexosamine. The polysaccharide moiety consisted of 63% glucose, 27% galactose, 7% mannose and 3% fucose. Fraction IN was found to increase the amount of superoxide anion in activated macrophages to 1.6-fold and the number of plaques in hemolytic plaque assay to 6-fold, respectively. These results indicate that the antitumor activity was exerted through immunopotentiation, but not through direct cytotoxicity against the tumor.

• Journal of Mushroom
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• v.12 no.3
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• pp.193-200
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• 2014

Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium were analyzed for their proximate composition, protein-bound polysaccharide, nucleic acid and amino acids. The content of ash and crude fiber in F. velutipes fruit body were higher than F. mycelium and C. militaris mycelium. C. militaris mycelium showed the highest crude fat content while F. velutipes fruit body had lowest. Nitrogen free extract content of the samples varied from 56.8% in F. velutipes fruit body to 61.9% in F. velutipes mycelium. The compositions of total protein and total free sugars of protein-bound polysaccharide were found to be significant differences for all samples. Nucleic acid related compounds were identified the 5'-GMP, 5'-XMP, 5'-IMP in all samples. The content of total nucleic acids were high in the orders of F. velutipes myclial (286.71 mg%), F. velutipes fruit body(187.36 mg%) and C. militaris mycelial(76.85 mg%). The highest content of 5'-GMP was found in F. velutipes fruit body. The most nucleic acid of F. velutipes mycelial and C. militaris mycelial were the 5'-XMP. As for the analysis of total amino acids, seventeen amino acids were identified by HPLC and the major amino acid was glutamic acid in all samples. The content of total amino acids were high in the orders of F. velutipes fruit body(19,919 mg%), F. velutipes mycelium(19,018 mg%) abd C. militaris mycelium(18,965 mg%). We determined the developing new food product such as amino acid drink and amino acid containing food using extracts of Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium.

• Journal of Chest Surgery
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• v.26 no.1
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• pp.47-53
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• 1993

The high relapse rate after curative surgery of lung cancer suggests that tumor cells are remained at the site of resection and in the distant organs. Postoperative radiochemoimmunotherapy including protein-bound polysaccharide PS-K[Copolang] and/or chemotherapy to improve the prognosis in lung cancer has been adopted. The patients with lung cancer who were treated with a combined modality therapy after surgery were reviewed to determine the effects of adjuvant immunotherapy[PS-K] and the relationship between midterm survival and clinicopathologic variables. During the past 5 years, 95 patients with lung cancer underwent resective operation. Of them, 30 cases were curative surgery, 29 were relative curative surgery, and the remainders were non-curative surgery. Postoperative combination therapies consisted of three types of therapies: postoperative BRM[biological response modifiers] with PS-K [Copolang] 50 mg/kg for 24 weeks[Group 1], chemoimmunotherapy with chemotherapy[a combination of cisplatin, etoposide, vindesine] and PS-K [Group 2], radioimmunotherapy with postoperative prophylactic irradiation to the mediastinum at total dose of 54 Gy-60 Gy and PS-K [Group 3] and surgery without adjuvant therapy[Group 4]. Twenty months survival rates of localized disease [Stages I and II] treated with PS-K, with radioimmunotherapy and no therapy were 73 %, 60 %, and 50 %, respectively [p [0.05]. Three-year survival rates of regionally advanced cases [stage Ilia and IIIb] were 23 % in Group 1.57 % in Group 2.20 % in Group 3, and 0 % in Group 4, respectively.According to above results, we suggest that postoperative combination therapy including PS-K might improve the prognosis of lung cancer. The similar survival pattern of patients with squamous cell carcinoma and adenocarcinoma treated with BRM, chemoimmunotherapy or radioimmunotherapy need to evaluate the role of postoperative immunotherapy[PS-K] in randomized studies.