• 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.

• BMB Reports
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• 제33권3호
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• pp.249-255
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• 2000

The Haemophilus influenzae type b (Hib) has been a major cause of bacterial meningitis in children who are less than two years old. The variable (V) region repertoire of adult Caucasian antibodies (Abs) to Hib polysaccharide (PS) has been characterized well. The majority of adult antibodies against Hib uses VL that is derived from the Vk gene A2 and have arginine at the N region. In order to explore the possibility those antibody responses to Hib-PS is variable in various age groups, we examined the VL regions of the antibodies to Hib-PS in Korean adults and children. We immunized Korean adults (n = 8) and children (n = 39) with Hib tetanus conjugated vaccines, isolated RNAs from the peripheral lymphocytes, and amplified the A2-derived VL regions by RT-PCR. The PCR products were subcloned and sequenced. Forty-seven out of 54 independent clones from children used the $J{\kappa}2$, or $J{\kappa}3$ gene in preference. The adults, however, used all of the $J{\kappa}$ genes evenly. With respect to the amino acid sequences of variable regions, adult $A2-J{\kappa}$ recombinants have a germline sequence. But, the 76th codon (AGC) of child $A2-J{\kappa}2$ recombinants was substituted with CGC (arginine) in most cases (88 %) and 77 percent of child clones using the $A2-J{\kappa}3$ genes have isoleucine-109 at the junction of $J{\kappa}-C{\kappa}$ instead of threonine that is found in a germline sequence. These results suggest that the mechanism of antibody production in young children is different from that of adults.

• 한국식품과학회지
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• 제25권4호
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• pp.299-306
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• 1993

본 연구에서는 일부 한국산 식품의 식이섬유 함량을 Englyst 등의 비색법과 gas chromatography 방법에 의한 비전분 다당류 분석에 의하여 측정하였다. Gas chromatography 방법에 의한 비전분다당류의 측정 결과, 곡류와 두류의 비전분다당류에서는 glucose와 5탄당인 arabinose 및 xylose, 미역, 다시마와 채소류의 불용성 비전분 다당류에서는 glucose, 수용성 비전분 다당류에는 uronic acid가 주요당이었다. 또한, 김의 불용성 및 수용성 비전분 다당류에는 galactose가 상당량 함유되어 있었다. Englyst와 Cummings의 비색법(x)과 gas chromatography 방법(y)에 의한 비전분다당류 값의 상관관계는 곡류, 두류 및 채소류의 식품군에서 y=1.01x-0.52(r=0.997, n=9)로 좋은 일치를 보여주었다. 한편 AOAC 공정법에 의한 총 식이섬유 함량(x)과 Englyst와 Cummings의 gas chromatography 방법에 의한 비전분 다당류 함량(y)의 상관관계 y=0.55x+2.14(r=0.786, n=12)로서 상관관계가 낮았으나, AOAC 공정법에 의한 총 식이섬유 함량(x)과 비전분 다당류 함량에 lignin 함량을 합한 값(y)의 상관관계는 y=0.68x+0.64(r=0.903, n=12)이었다.

• The Plant Pathology Journal
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• 제33권3호
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• pp.276-287
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• 2017

RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii.

• Preventive Nutrition and Food Science
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• 제7권2호
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• pp.139-145
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• 2002

Hen egg-white lysozyme, ovalbumin, egg-yolk phosvitin, acid-precipitated soy protein and $\alpha$$_{sl}$ milk casein were covalently linked with galactomannan through a controlled dry-heating at 6$0^{\circ}C$ under 79% relative humidity without any chemical reagent. Neoglycosylation by the covalent binding of polysaccharide chains brought a significant improvement into the surface functionalities of food proteins. Excellent emulsifying properties and foaming properties were observed in all protein-galactomannan conjugates. Bacterial mutagenesis tests and animal dose test were done to evaluate the food safety of the protein-galactomannan conjugates. The neo-glycoproteins were negative for Ames test using Salmonella typhimurium TA100 (hisG46) and TA98 (hisD3052) strains, and rec-assay using Bacillus subtilis Hl7 (rec) and M45 (re $c^{+}$) strains. All substances were also nontoxic for oral administration to rats. L $D_{50}$ 's of these substances were all more than 7.5 g/kg body-weight of rat. No effect was also observed in the weight increases and the concentrations of total cholesterol, triglyceride and phospholipids in blood serum of the administrated rats with 7.5 g/kg conjugates. Thus, Maillard-type protein-polysaccharide conjugates prepared by covalent attachment of galactomannan to food proteins were proposed to be useful as a safe functional biopolymer in this study.y.

• Applied Biological Chemistry
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• 제39권2호
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• pp.159-164
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• 1996

혈액 항응고 활성물질을 검색하기 위하여 73종의 한약재, 41종의 산채류 및 일반 채소류, 5종의 해조류들의 열수추출물들에 대해 혈액응고 지연시간(Tr)을 시험하였다 1차검색에서 비교적 활성이 높았던 택사, 삼칠근, 현호색, 운지, 마늘, 청각의 산성 수추출물에서 가장 높은 항응고 활성(대조군에 비해 4.3배)을 보였다. 청각(1.2KG)을 0.8% 염산 수용액(24l)으로 추출한 후 메탄을 환류, 에탄올 환류, 침전 및 투석하여 활성이 약 2배 증가된 조다당 CF-1을 얻었다. CF-1은 80.8%의 다당과, 8.7%의 단백질 및 13.3%의 황산기로 구성되어 있었으며, 다당류는 arabinose, galactose, 및 glucose가 주구성당으로 확인되었다. 이 조다당 CF-1의 항응고 활성 본체를 확인하기 위해 pronase처리와 periodate 산화를 행한 결과 pronase 처리시에는 활성의 변화가 없었으나 periodate 산화시에는 약 70%로 활성이 감소한다는 사실로부터 CF-1의 항응고 활성은 다당에 의해 기인됨을 알 수 있었다.

• Food Science and Biotechnology
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• 제16권6호
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• pp.928-934
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• 2007

Physiochemical properties, such as yield and molecular weight distribution of polysaccharide fractions, of polysaccharides in the enzymatic hydrolysates of purslane were investigated and characterized. A higher amount of micro nutrients, such as potassium (9,413 mg/100 g), phosphorus acid (539 mg/100 g), leucine, alanine, lysine, valine, glycine, and isoleucine, was present in whole purslane. The yield of water soluble polysaccharides (WSP) was 0.29, 7.01, and 7.94% when extracted using room temperature water (RTW), hot-water (HW), and hot temperature/high pressure-water (HTPW), respectively, indicating that HW or HTPW extraction may be effective to obtain WSP from purslane. The average ratio of L-arabinose:D-galactose in the WSP was 37:49, 34:37, and 27:29, when extracted using RTW, HW, and HTPW, respectively. These results indicate that water was a suitable extraction solvent for preparation of the arabinogalactan component of whole purslane. A higher yield and total carbohydrate content was obtained by using Viscozyme L instead of Pectinex 5XL during extraction of the WSP, which indicates that enzymatic treatment of purslane may be an effective method to control the Mw of polysaccharides. Finally, it was confirmed that Viscozyme L is a suitable enzyme for the hydrolysis and separation of polysaccharides obtained from purslane.

• 통합자연과학논문집
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• 제3권2호
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• pp.72-77
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• 2010

Alginate lyases, also known as alginases or alginate depolymerases, catalyze the degradation of alginate by a ${\beta}$-elimination mechanism that has yet to be fully elucidated. Alginate is a copolymer of ${\alpha}$-L-guluronate (G) and its C5 epimer ${\beta}$-D-mannuronate (M), arranged as homopolymeric G blocks, M blocks, alternating GM or random heteropolymeric G/M stretches. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a ${\beta}$-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. Till now there is no crystal structure available for this class of proteins. Since there is no template with high sequence identity, three-dimensional coordinates for exotype alginate lyase (PL 15 family) were determined using modeling methods (Comparitive modeling and Fold recognition). The structures were modeled using the X-ray coordinates from Heparinase protein family (PDB code: 3E7J). This enzyme (Atu3025) displays enzymatic activity for both poly-M and poly-G alginate. Since poly-M is widespread; docking of a tri-mannuronate against the modeled structure was performed. We identified some of those residues which are crucial for lyase activity. The results from this study should guide future mutagenesis studies and also provides a starting point for further proceedings.

• 상하수도학회지
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• 제25권5호
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• pp.681-689
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• 2011

Submerged membrane bio-reactor (SMBR) has several advantages such as high MLSS, long SRT, and low F/M ratio at wastewater treatment. So, this has widely applied over the world and many studies have been conducted. However, membrane fouling remains an inevitable problem. This study was investigated using bench-scale SMBR with three poeration modes. Raw waters were prepared by addition of starch, acetic and fibric acid to recovery water of zeolite. The efficiency of nitrification and COD were very stable as about 95% and 80%, respectively. And critical flux was 128.8L/$m^{2}$/hr. The result of biodegradability test was following values at the each mode : Ss+Xs/$C_{T}$=81.7%, 35.1% and 45.3%, $X_{I}+S_{I}/C_{T}=18.3%$, 64.9% and 54.7%. When particulate matters such as $X_{I}$ and $X_{S}$ in influent are increased, membrane fouling will take place more and more. A relative ratio of filtration resistance to the fouling occurred by the cake layer was increased when increased the portion of $X_{I}$ and polysaccharide. It was thought that the formation of cake layer was promoted due to bond between $X_{I}$ and vicid material s generated from the polysaccharide.

• Archives of Pharmacal Research
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• 제20권2호
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• pp.128-137
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• 1997

The effects of cylindan, a polysaccharide isolated from the basidiocarps of Agrocybe cylindracea, on murine sarcoma 180 tumor and murine immune cells were examined after intraperitoneal administration. Cylindan exhibited a marked life extension effect in mice against ascite forms of sarcoma 180 and Lewis lung carcinoma at a dose of 50 mg/kg/day, although it did not show any direct cytotoxicity against sarcoma 180, X5563, and MM46 murine tumor cells. Cylindan increased numbers of bone marrow stem cells as well as peritoneal exudate cells in flow cytometry using monoclonal antibodies. The tumor bearing mice group apparently showed the increase of macrophages and cytotoxic T lymphocytes in mouse spleen cells during the early stage of tumor growth. But during the later stage, the control group decreased immune cells and cylindan restored the decreased immune cells in the tumor bearing mice to the normal level. In non-specific immune response, cylindan stimulated the bacterial phagocytosis and acid phosphatase production in macrophages. It also activated components of the alternative complement pathway and natural killer activity against YAC-1 lymphoma. In number of plasma cells as token of stimulation of the differentiation of B lymphocytes. In cellular immunity, cylindan restored the depressed response of delayed type hypersensitivity in the tumor bearing mice to 60% of the normal level and increased the interleukin-2 (IL-2) responsiveness in the IL-2 dependent CTLL-2 cells. These results suggest that cylindan did not show direct cytotoxic effects on tumor cells but restored the decreased immune response of the tumor bearing mice.