• KSBB Journal
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• 제24권1호
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• pp.59-64
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• 2009

The callus formation from inferior leaf of Aloe saponaria was induced in M & S medium supplemented with 10-30 ${\mu}M$ NAA (${\alpha}$-naphthalene acetic acid) and 3-7 ${\mu}M$ kinetin under incubation in the dark at $25^{\circ}C$ for 6 weeks. The hot water extract ($100^{\circ}C$, 24 hrs) from cultured callus was obtained and the components analysis for the extract were examined to determine the callus can synthesized the bioactive component such as Aloe polysaccharide. The freeze dried extract contained the sugar of 53.2%, protein of 7.3%, ash of 18.5% and water of 21% (w/w). Two fractions (Fr-I and Fr-II) were obtained by Sepharose CL-4B gel permeation chromatography and Fr-I, major fraction was further purified with dialysis. From sugar analysis by TLC and GC, the purified Fr-I fraction consisted of glucose (77.6%), galactose (17.7%), mannose (4.7%, w/w) and uronic acid (trace). The molecular weight of purified Fr-I fraction determined by GPC was about 110 kDa.

• 한국식품영양학회지
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• 제28권5호
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• pp.821-828
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• 2015

ASW0 is a polysaccharide derived from the perennial herb Aster scaber Thunberg. We isolated ASW0, a fraction of crude polysaccharide, by means of ethanol precipitation and dialysis after hot water extraction to investigate its physicochemical properties and immunostimulatory effects. ASW0 contains neutral sugar (45.7%), acidic sugar (51.6%), protein (2.3%), and 2-keto-3-deoxy-D-manno-octonate (KDO) (0.4%). The neutral sugar in ASW0 (in mole percentage) was mainly composed of arabinose (34.5 mol%), glucose (31.1 mol%), galactose (14.9 mol%), and rhamnose (8.1 mol%), which are characteristic of pectic polysaccharides. ASW0 also contained small amounts of xylose, mannose, and fucose. The anti-complementary activity of ASW-0 was similar to that of polysaccharide K (used as positive control). ASW0 exhibited no cytotoxicity in RAW 264.7 macrophages and dramatically increased nitric oxide (NO) production in a dose dependent manner ($0.3{\sim}30{\mu}g/mL$). Also, macrophages stimulated with ASW0 showed enhanced production of immunostimulatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor alpha ($TNF-{\alpha}$) in a dose dependent manner. These results suggest that the ASW0 have a potent immunostimulatory effect and can be used as a natural immune health ingredient.

• 한국식품영양과학회지
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• 제33권7호
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• pp.1092-1097
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• 2004

큰느타리버섯의 기능성 식품으로서 활용도를 높이기 위해 동결 건조된 자실체에서 분리한 조다당체 추출물이 면역세포 활성에 미치는 효과를 조사한 결과는 다음과 같다. 조다당체 추출물은 300 및 1,000 $\mu$g/mL 농도에서 비장세포의 증식을 유도하였으며, 이 때 비장세포는 IL-6와 IFN-${\gamma}$ 분비를 유도하는 것으로 나타났다 조다당체 추출물은 농도 의존적으로B세포의 증식을 유도하였으며, 특히 100 $\mu$g/mL농도 이상에서는B세포의 증식이 현저히 증가하는 것으로 나타났다. 그리고 조다당체 추출물 1,000 $\mu$g/mL 농도에서 B세포가 생산하는 IgGl, IgG2a, IgG3의 분비량이 현저히 증가하였다. 또한 농도 의존적으로 대식세포주의 일산화질소 생산을 유도하였으며, 대식세포가 분비하는 IL-6, TNF-$\alpha$, GM-CSF의 생산도 현저히 증가하는 것을 확인할 수 있었다.

• ALGAE
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• 제26권4호
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• pp.343-350
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• 2011

Fermentation and enzyme-assisted extraction (EAE) improve nutritional and functional properties of foods by increasing the extraction of active compounds, ingestion rates, and body absorption. In this study, we investigated whether applying the EAE process improves the extraction and isolation efficiency of a polysaccharide from fermented Ecklonia cava (FE), which inhibited NO production in lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results showed that the FE using the fungi Candida utilis and two different bacteria, namely Lactobacillus brevis and Saccharomyces cerevisiae increased protein and carbohydrate contents in comparison with those in non-fermented E. cava (NE). Aqueous extracts of fermented E. cava increased extraction yields and carbohydrate content, compared with the aqueous extract of NE. In addition, treating LPS-stimulated RAW 264.7 cells with aqueous extracts resulted in reduced NO production compared to that in LPS-treated cells. Ten EAEs of L. brevis-fermented E. cava (LFE) improved NO inhibitory effects in LPS-activated RAW 264.7 cells and the Viscozyme extract (VLFE) from the resulting extracts showed the highest NO inhibitory effect. We found that the >30 kDa fraction of VLFE led to markedly high inhibition of LPS-induced NO production as compared to that in the <30 kDa fraction. The crude polysaccharide isolated from >30 kDa fraction (VLFEP) consisted of fucose and markedly decreased NO production induced by LPS stimulation. VLFEP could be useful as an anti-inflammatory agent to suppress macrophage activation.

• Food Science and Biotechnology
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• 제16권4호
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• pp.600-604
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• 2007

In this study, we first prepared 3 kinds of powders with different molecular masses from the crude protein-bound polysaccharide extract of Agraricus blazei Murill through ultrafiltration, followed by spray-drying. Then, the antitumor activities of the powders were analyzed. Size exclusion chromatography coupled with a multi-angle laser-light-scattering system showed the 3 powders had the following molecular ranges: below 10 kDa (SD-1), 10 to 150 kDa (SD-2), and above 150 kDa (SD-3), representing peak molecular weights of $8.26{\times}10^3,\;9.65{\times}10^4$, and $5.94{\times}10^6\;g/mol$, respectively. All the powders stimulated macrophage RAW264.7 cells to produce nitric oxide, of which SD-2 and SD-3 were superior to the crude extract powder (CP-SD), while SD-1 showed the lowest activity. Similar results were found for their cytotoxicities against human cancer cell lines (A549, MCF-7, and AGS), where the highest activity was obtained with the SD-2 treatment for 72 hr at $1,000\;{\mu}g/mL$. The MCF-7 cell line was less sensitive to the powders than the other cells. From this research we found that ultrafiltration, in combination with spray-drying, is applicable for preparing protein-bound polysaccharide powders with higher antitumor activities.

• 한국식생활문화학회지
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• 제37권5호
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• pp.438-445
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• 2022

Laminaria japonica is a type of brown algae widely consumed in Asian countries and contains many essential nutrients and exhibits anti-obesity, antioxidant, and anti-inflammatory effects. In this study, the antioxidant and immunomodulatory effects of a Laminaria japonica water extract (LJE) were investigated using an in vitro model. Mean total polyphenol content of LJE was 2.16±0.11 ㎍ GAE/mg, and LJE dose-dependently inhibited ABTS radical activity but did not scavenge DPPH radicals. In addition, LJE enhanced nitric oxide (NO) production and upregulated the mRNA expressions of proinflammatory cytokines (i.e., tumor necrosis factor-α and interleukin-6) in RAW 264.7 cells. On the other hand, LJE inhibited NO production and downregulated proinflammatory cytokine mRNA levels in endotoxin-stimulated RAW 264.7 cells. Thus, our data show that LJE has moderate antioxidant activity and biphasic immunomodulatory effects on RAW 264.7 cells. In summary, the study indicates that LJE has potential therapeutic use as a novel biphasic immuno-modulator.

• Journal of Microbiology and Biotechnology
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• 제31권6호
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• pp.855-866
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• 2021

The effects of various carbon sources on mycelial growth and polysaccharide synthesis of the medicinal fungus Inonotus obliquus in liquid fermentation were investigated. After 12-d fermentation, mycelial biomass, polysaccharide yield, and polysaccharide content were significantly higher in Glc+Lac group (glucose and lactose used as combined carbon source) than in other groups. Crude polysaccharides (CIOPs) and the derivative neutral polysaccharides (NIOPs) were obtained from mycelia fermented using Glc, fructose (Fru), Lac, or Glc+Lac as carbon source. Molecular weights of four NIOPs (termed as NIOPG, NIOPF, NIOPL, and NIOPGL) were respectively 780.90, 1105.00, 25.32, and 10.28 kDa. Monosaccharide composition analyses revealed that NIOPs were composed of Glc, Man, and Gal at different molar ratios. The NIOPs were classified as α-type heteropolysaccharides with 1→2, 1→3, 1→4, 1→6 linkages in differing proportions. In in vitro cell proliferation assays, viability of RAW264.7 macrophages was more strongly enhanced by NIOPL or NIOPGL than by NIOPG or NIOPF, and proliferation of HeLa or S180 tumor cells was more strongly inhibited by NIOPG or NIOPGL than by NIOPF or NIOPL, indicating that immune-enhancing and anti-tumor activities of NIOPs were substantially affected by carbon source. qRT-PCR analysis revealed that expression levels of phosphoglucose isomerase (PGI) and UDP-Glc 4-epimerase (UGE), two key genes involved in polysaccharide synthesis, varied depending on carbon source. Our findings, taken together, clearly demonstrate that carbon source plays an essential role in determining structure and activities of I. obliquus polysaccharides by regulating expression of key genes in polysaccharide biosynthetic pathway.

• 한국균학회지
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• 제27권1호통권88호
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• pp.82-86
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• 1999

영지버섯(Ganoderma lucidum WK-003) 균사체의 액체 배양에 의하여 세포외 생물고분자 물질의 산업적 생산을 위한 최적 조건을 검토하였다. 세포외 고분자물질 생산을 위한 산업용배지의 최적 농도는 molasses 5%와 CSL 2.5%에서 균사체 생장(12.4 g/l)과 균체외 다당체 생산(3.6g/l)이 최대를 나타났으며, 5 l 발효조 배양시에는 균사체 생장이 합성배지(8.1g/l)일때가 산업용배지(7.3 g/l)보다 높게 나타났으나, 균체외 다당체 생산은 합성배지(7.2 g/l)보다 산업용 배지(11.2 g/l)일 때가 더 높게 나타났다. 간 보호 효과는 합성배지와 최적화된 산업용 배지에서 생산된 균체외 다당체를 $CCl_4$ 독성이 유발된 rat에 경구 투여하였을 때 주요 지표를 나타내는 GPT활성은 산업용배지에서 생산된 균체외 다당체 인 경우 704IU/L에서 356IU/L로 낮추었으며, 합성배지에서 생산된 균체외 다당체는 704IU/L에서 349IU/L로 낮춘 결과를 보였다. 따라서 합성배지와 산업용 배지에서 생산된 균체외 다당체의 간 보호 작용은 거의 차이가 없는 것으로 나타났다.

• 한국균학회지
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• 제37권1호
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• pp.91-95
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• 2009

자연에서 채집한 치마버섯 자실체로부터 분리한 Schizophyllum commune 균을 배양하여 얻은 균사체 배양물로부터 추출한 다당류의 물리화학적 특성에 대하여 조사하였다. 액체배양 균사체로부터 추출된 다당류는 방법에 따라 $0.19{\sim}1.63%$의 수율을 나타내었다. 이들 다당류의 총 당은 $48.9{\sim}72.0%$이고, 단백질은 $15.3{\sim}32.0%$를 함유하고 있는 것으로 나타났다. 추출한 SC-EP 다당류의 구성 당은 glucose가 69.6%로 주를 이루고, fructose가 26.1%, 소량의 xylose로 구성된 것으로 나타나는 것으로 보아 hetero 다당류인 것으로 추정되었다. 또한, 산성 아미노산인 aspartic acid, glycine 및 glutamic acid 등은 각각 12.5%, 18.1% 및 15.2%, 중성아미노산인 alanine은 9.9%, 당과 단백질의 O${\beta}-glycosidic$ 결합에 관여하고 있는 threonine과 serine의 경우는 2.9% 및 2.7%로 나타났다. IR 분석결과 이들은 ${\alpha}$ 와 ${\beta}$-glucan이 혼재된 다당류인 것으로 나타났다. 한외여과장치로 제조한 다당류(SC-UP)의 0.5% 용액의 점도는 20 rpm 시 70 cps로 나타났다. 이상의 치마버섯 액체배양 균사체를 열수추출하여 에탄올 침전시켜 얻은 다당류는 당과 단백질이 결합된 단백다당류의 결합 형태를 이루고, 당은 2가지 이상의 당이 혼재하고 있으며, 아미노산은 산성아미노산의 함량이 높은 것으로 나타났다. 다당류의 결합 양식은 ${\alpha}$ 와 ${\beta}$-glucan이 혼재하고 있으며, 강한 점성을 나타내었다.

• Applied Biological Chemistry
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• 제3권
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• pp.25-48
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• 1962

tragacanth gum의 화학구조(化學構造)를 구명(究明)하기 위(爲)하여 미국(美國) 약전(藥典)의 tragacanth gum 분말(粉末)을 가지고 다음의 실험(實驗)을 통(通)하여 이의 성분(成分)의 하나인 polysaccharide C를 분리(分離)하여 이의 화학구조(化學構造)를 밝혔다. (1) tragacanth gum을 85% 주정(酒精)으로 처리(處理)해서 중성다당류(中性多糖類)로 polysaccharide C를 분리(分離)하였으며 구성당(構成糖)으로 L-rhamnose, D-xylose, L-arabinose 및 D-galactose를 paper chromatography와 Cellulose column chromatography로 분리(分離), 동정(同定)하였다. 이의 molar ratio는 2:1:17:9이며 비선광도(比旋光度)는 $[{\alpha}]^{30}_D-72.2이다. (2) 구성당(構成糖)의 결합위치(結合位置)를 구명(究明)하기 위(爲)해 Hawarth 법(法)과 Purdietldir(試藥)을 가지고 methyl화(化)시켜 methyl화(化) polysaccharide C를 얻었으며 비선광도(比旋光度) $[{\alpha}]^{22}_D-102를 보였다. 이것을 가수분해(加水分解)시켜 paper chromatography와 column chromatography를 통(通)해 methyl화단당(化單糖)으로 1,3,5-tri-O-methyl-L-arabofuranose, 3,4-di-O-methyl-L-rhamnose, 2,3-di-O-methyl-D-xylose, 2,3,4-tri-O-methyl-D-galactopyranose, 2,4-di-O-methyl-L-arabopyronose, 2,4-di-O-methyl-D-galactose, 2-O-methyl-L-arabinose 및 L-arabinose를 분리(分離), 동정(同定)하였다. (3) 산(酸)의 각종농도(各種濃度)에 따른 부분적(部分的) 가수분해(加水分解)를 시켜 polysaccharide C의 end group, 측지(側枝) 또는 주쇄(主鎖)를 이루는 구성당(構成糖)을 밝히기 위(爲)하여 0.05 N-HCl로 제1차(第一次) 가수분해(加水分解). 0.1N-HCl로 제2차(第二次) 가수분해(加水分解), 0.3N-HCl로 제3차(第三次) 가수분해(加水分解)를 하여 가수분해물(加水分解物)과 비가수분해물(非加水分解物)에서 각각(各各) 다음과 같은 구성단당(構成單糖)을 검출(檢出)하고 이들의 molar ratio를 측정(測定)하였다. 제1차(第一次) 가수분해물(加水分解物)(A)에서 L-arbinose, 비가수분해물(非加水分解物)(A')에서 L-rhamnose, D-xylose, L-arabinose 및 D-galactose; 제2차(第二次) 가수분해물(加水分解物)(B)에서 L-arabinose와 D-galactose, 비가수분해물(非加水分解物)(B')에서 L-rhamnose, D-xylose, L-arabinose, 및 D-galactose; 제3차(第三次) 가수분해물(加水分解物)(C)에서 L-rhamnose, D-xylose, L-arabinose 및 D-galactose, 비가수분해물(非加水分解物)(C')에서 D-xylose와 D-galactose를 검출(檢出)하였다. (4) 구성당(構成糖)의 형태(形態)와 구조(構造)를 밝히기 위(爲)해 polysaccharide C에 대한 periodate산화(酸化) 실험(實驗)을 하여 $C_5H_8O_4$당(當) periodate의 소비(消費)와 formic acid의 생성량(生成量)을 측정(測定)하였는데 periodate의 소비량(消費量)은 1.23 mole, formic acid의 생성량(生成量)은 0.78 mole이다.