• Applied Biological Chemistry
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• 제41권1호
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• pp.60-66
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• 1998

10종의 식용 버섯 자실체에 대한 항보체 활성 검색에서 가장 높은 활성을 나타낸 양송이버섯 자실체를 대상으로 항보체 활성물질의 추출조건을 조사하였다. 최적조건으로서 5% urea가 함유된 0.1 N NaOH 용액으로 $65^{\circ}C$에서 2시간동안 양송이버섯 자실체를 추출한 후 methanol 가용획분이 제거된 조다당 획분 AB-0를 얻었다. AB-0는 1 mg/ml에서 90% 이상의 높은 항보체활성을 나타내었으며 마우스에 이식된 sarcoma-180에 대하여 74%의 저지효과를 보였다. AB-0를 acetone 농도에 따른 침전 분획을 실시하여 AB-20, AB-40, AB-60, AB-80, AB-A의 5개의 획분을 얻었다. 이중 가장 높은 항보체활성과 수율을 나타낸 AB-20 획분은 탄수화물 39%, 단백질 46%를 함유하였으며, 구성당은 glucose, arabinose, xylose, galactose, mannose가 6.49 : 1.98 : 1.24 : 1.00 : 0.71의 비율로 존재하였고 구성아미노산은 isoleucine(12.60%), glutamic acid+glutamine(12.45%), valine(11.79%), alanine(11.46%), leucine(10.19%)와 aspartic acid, asparagine(10.56%) 등 이었다. Pronase 처리한 AB-20에서는 78.5%의 항보체활성을 나타내고 대조군에 비해 14.5% 감소하였으나, AB-20의 periodate 산화물에서는 활성이 42.6%로 50% 이상 크게 감소하였다. 따라서 양송이 버섯의 알칼리 추출물중 가장 강한 활성을 보인 AB-20의 주요 활성부위는 다당류이며 단백질도 일부 관여하는 것으로 생각되었다.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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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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• 제9권4호
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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.

• 한국식품영양과학회지
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• 제27권6호
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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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• 제2권2호
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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.

• 한국균학회지
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• 제10권4호
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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.

• 한국균학회지
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• 제13권3호
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• pp.131-139
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• 1985

들버섯의 배양 균사체로부터 추출한 단백다당체는 마우스에 이식된 sarcoma 180에 대해서 유효한 항암력을 나타내었다. DEAE-Sephadex A-50을 이용하여 정제한 결과 4개의 분획을 얻었다. 이중 가장 항암효과가 큰 분획 C는 56.1 %의 종양저지율을 나타내었다. 분회 C는 45 %의 다당류와 18 %의 단백질로 되어있다. 그 다당류는 mannose(42.0 %), glucose(25.5 %), xylose(16.6 %), fucose(11.1 %)와 galactose(4.8 %)로 되어 있고 단백질은 17종의 아미노산으로 되어 있었다. 들버섯의 항암성분은 마우스 복강내에 있는 macrophage의 수를 현저히 증가시켰으며 비장내의 용혈반형성 세포수도 증가시킴으로써 면역 증강 작용이 있다.

• Food Science and Biotechnology
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• 제18권6호
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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.

• 한국균학회지
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• 제22권2호
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• pp.184-189
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• 1994

구름 버섯균(C. versicolor)를 고체배양(solid-substrate culture)하여 단백다당류를 생산하기 위한 각종조건을 검토하였다. 여러가지 고체재료가 이용될 수 있으나 배양 후 단백다당류의 열수추출 과정에서의 부적합성 등의 이유로 인하여 실질적으로 zeolite나 난 재배용 토양이 이용 가능한 천연고체 재료라고 볼 수 있다. Zeolite와 난 재배용 토양을 이용한 고체발효에서 단백다당류 생산에 미치는 탄소원의 영향을 검토한 결과 glucose, sucrose 및 starch에서 단백다당류 생산량이 높게 나타났으며, 질소원중에서는 bactosoytone과 peptone이 고체발효에 의한 단백다당류 생산에 가장 적합한 질소원으로 확인되었다. 단백다당류 생산을 위한 고체발효에서 최적 pH범위는 5-6이었고, 고체재료 100 g당 조단백다당류 생산량이 5-6 mg에 이르렀다. 배지조성별 단백다당류의 생산성 시험의 결과 C. versicolor의 최적배지 조성인 CVM에서 최고의 생산성을 보였으며, 종합적으로 균사성장에 적합한 조건에서 단백다당류의 생산성도 높다고 하겠다.

• 한국식품저장유통학회지
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• 제11권1호
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• pp.47-52
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• 2004

η$_{evap}$분자량에 따라 다양한 생리활성을 나타내는 아가리쿠스버섯에서 분리한 조단백다당류를 고부가가치 기능성 소재로 개발하기 위하여 막분리 및 분무건조 공정에 따른 특성을 조사하였다. 아가리쿠스버섯 조단백다당류를 분자량 크기에 따라 여과하는 동안 공정온도에 따른 투과플럭스의 변화는 10 kDa와 130 kDa 모두에서 유사하였는데, 막분리 온도 및 압력이 증가함에 따라 비례적으로 증가하였으며, 4$0^{\circ}C$에서의 투과플럭스가 가장 높았다. 시간에 따른 투과플럭스의 변화는 막분리 30분 경과시 급속히 감소하였으며, pore size가 10 kDa와 150 kDa일 때 공정압력이 각각 1.2kgf/$\textrm{cm}^2$와 2kgf/$\textrm{cm}^2$에서 가장 높았다. 한외여과 후 아가리쿠스버섯 조단백다당류의 점도는 여과 전 13.9cP에서 여과후 10kDa이 하와 10∼150 kDa구간은 각각 10.8, 11.9cP로 점도가 저하되었으나, 150 kDa이상은 20.1cP로 점도가 증가되었다. 분자량 크기가 다른 3가지 분획물에 대한 분무건조 공정에서의 열효율성은 가열공기온도, 시료공급속도 및 농도가 증가할수록 열효율성이 증가함을 알 수 있었으며, 시료공급속도가 가장 중요한 영향인자임을 확인하였다.다.