• Journal of Microbiology and Biotechnology
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• 제20권7호
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• pp.1053-1060
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• 2010

Cordyceps militaris, an entomopathogenic fungus belonging to the class Ascomycetes, has been reported to have beneficial biological activities such as hypoglycemic, anti-inflammatory, antitumor, antimetastatic, hypolipidemic, immunomodulatory, and antioxidant effects. In this study, the crude water-soluble polysaccharide CMP, which was obtained from the fruiting body of C. militaris by hot water extraction and ethanol precipitation, was fractionated by DEAE-cellulose and Sepharose CL-6B column chromatographies. This process resulted in three polysaccharide fractions, termed CMP Fr I, CMP Fr II, and CMP Fr III. Of these fractions, CMP Fr II, with an average molecular mass of 127 kDa, was able to upregulate effectively the phenotypic functions of macrophages such as NO production and cytokine expression. The chemical property of the stimulatory polysaccharide, CMP Fr II, was determined based on its monosaccharide composition, which consisted of glucose (56.4%), galactose (26.4%), and mannose (17.2%). Its structural characteristics were investigated by a combination of chemical and instrumental analyses, including methylation, reductive cleavage, acetylation, Fourier transform infrared spectroscopy (FTIR), and gas chromatography-mass spectrometry (GCMS). Results indicated that CMP Fr II consisted of the (1${\rightarrow}$4) or (1${\rightarrow}$2) linked glucopyranosyl or galactopyranosyl residue with a (1${\rightarrow}$2) or (1${\rightarrow}$6) linked mannopyranosyl, glucopyranosyl, or galactopyranosyl residue as a side chain. The configuration of the ${\beta}$-linkage and random coil conformation of CMP Fr II were confirmed using a Fungi-Fluor kit and Congo red reagent, respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.351-358
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• 2007

Carbohydrate-protein interactions are known to be important in gamete interactions. Several evidence indicated that a fucose-containing sulfated polysaccharide fucoidan was potential inhibitor of fertilization in vitro and thus fucose seemed to be part of the recognition signal of gamete interaction in mammals. In recent investigation we found that ${\alpha}$-L-fucosidase activity was present in boar spermatozoa and it was related to sperm binding to and penetration into zona pellucida (ZP) in vitro. The objective of this study was to determine the effects of monosaccharide L-fucose and polysaccharide fucoidan on sperm ${\alpha}$-L-fucosidase activity and relation to sperm-oocyte interaction in pig. Results indicated that the activity of sperm ${\alpha}$-L-fucosidase was largely inhibited (62%) when sperm suspension was treated with monosaccharide L-fucose. It also significantly inhibited the number of sperm binding to ZP (32%) and penetration into zona-intact oocytes (72%), but did not inhibit penetration into zona-free oocytes when fertilization medium contained L-fucose. The chlorotetracycline (CTC) assessment showed that L-fucose did not affect induction of sperm capacitation and acrosome reaction. In contrast, the activity of sperm ${\alpha}$-L-fucosidase was not inhibited when sperm suspension was treated with polysaccharide fucoidan but sperm-ZP binding was greatly inhibited (85%) and completely blocked sperm penetration into zona-intact or zona-free oocytes. The CTC assessment showed that fucoidan increased the F pattern and decreased the AR pattern sperm. These results suggested that the different inhibitory mechanisms were present between monosaccharide L-fucose and polysaccharide fucoidan on sperm-oocyte interaction, the inhibition effect of ${\alpha}$-L-fucose on sperm binding and penetrating into ZP caused sperm ${\alpha}$-L-fucosidase inhibited by ${\alpha}$-L-fucose.

• KSBB Journal
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• 제10권5호
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• pp.518-524
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• 1995

해양으로부터 polysaccharide를 생산하는 세균을 분리하여 동정한 결과 Zoogloea sp.(KCCM 10036) 로 밝혀졌으며, 이 해양유래 Zoogloea sp.는 셔로 다 른 두종류의 다당류인 수용성다당류(WSP)와 세포 결합다당류(CBP)를 통시에 생산하여 지금까지 보 고된 관련 미생물과는 다른 특이성을 보였다. 이들 중 WSP는 원심분리 시킨 배양액의 상등액으로부터 acetone 침전법에 의해 분리하였고, CBP는 침전물 로부터 acetone 빛 cetylpyridinium chloride 처 리 에 의 해 분리하였다. WSP와 CBP는 모두 단백 질을 함유하였으나, 이 중 CBP가 보다 높은 단백질 함유 율을 나타내었다. WSP를 다량 생산하기 위한 최척 탄소원은 fructose였고, CBP의 경우는 glucose였 다. Polysaccharide 생산촉진인자로 biotin, ampicillin, 그리고 surf actant를 각각 배지에 첨가하였을 경우, 각각의 농도가 $0.2mg/\ell, 0.4mg/\ell,0.6mg/\ell,$에서 생산량이 가장 높은 값을 보였다. 1%의 glucose를 탄소원으로 하여 fermentor 배양하였을 때, $9.1g/\ell$ 의 WSP와 $2.5g/\ell$ 의 CBP가 생산되었다.

• 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.

• Journal of Plant Biotechnology
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• 제4권3호
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• pp.109-115
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• 2002

Submerged cultures of Ganoderma lucidum, a valuable mushroom in traditional Chinese medicine, were used for production of bioactive Banoderic acids and Ganoderma polysaccharides. The significant effects of oxygen supply were demonstrated in both shake flasks and bioreactors. By changing the medium loading volume in a shake flask, a different value of initial volumetric oxygen transfer coefficient ($K_L$a) was obtained, and a higher $K_L$a value led to a higher biomass density and a higher productivity of both intracellular polysaccharide and ganoderic acid. In a stirred bioreactor, at an initial $K_L$a of 78.2 $h^{-1}$, a maximal cell concentration of 15.6 g/L by dry weight was obtained, as well as a maximal intracellular polysarcharide (IPS) production of 2.2 g/L and its maximal productivity of 220 mg/(L.d). An increase of initial $K_L$a led to a higher production and productivity of GA, and the GA production and productivity at an initial $K_L$a of 96.0 $h^{-1}$ was 1.8-fold those at an initial $K_L$a of 16.4 $h^{-1}$. The fundamental information obtained in this study may be useful for efficient large-scale production of these valuable bioactive products by the submerged cultures.

• BMB Reports
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• 제42권12호
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• pp.794-799
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• 2009

The inhibition of nitric oxide (NO) and cyclooxygenase-2 (COX-2) production is considered to be a promising approach to the treatment of various diseases, including inflammation and cancer. In this study, we examined the effects of the Agrocybe chaxingu $\beta$-glucan (polysaccharide) on lipopolysaccaride (LPS)-induced nitric oxide (NO) and cyclooxygenase-2 (COX-2) expression in murine macrophage Raw 264.7 cells as well as 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear edema in mice. The polysaccharide significantly inhibited (P < 0.01) LPS-induced iNOS and COX-2 expression levels in the cells. Furthermore, topical application of polysaccharide resulted in markedly inhibited (P < 0.01) TPA-induced ear edema in mice. These results suggest that this polysaccharide may be used for NO- and COX-2-related disorders such as inflammation and cancer.

• 미생물학회지
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• 제33권2호
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• pp.142-148
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• 1997

제한통성 메탄올 자호세균인 Methylovorus sp. strain SS1은 최적 성장 조건하에서는 소량의 세포외 다당류(EPS)를 생산하였지만, 질소원이 결핍된 성장 조건하에서는 성장속도는 느렸지만 다량의 EPS를 생산하였다. EPS는 배지내의 탄소대 질소 비율이 5.2일 때 가장 많이 생산되었다. EPS 생산을 위한 최적 온도는 30^{\circ}C.$이고 최적 pH는 6.5였다. EPS는 탄수화물과 단백질 및 약간의 피루브산으로 구성되어 있었고, 환원당으로는 다량의 포도당과 소량의 mannose가 존재하였다. 에탄올을 처리한 EPS(EPSae)에는 에탄올을 처리하지 않은 EPS(EPSbe)에 존재하던 피루브산이 존재하지 않았고, EPS보다 단백질의 양도 적고 점성도 낮았다. EPSbe의 점성은 NaCl에 의해 큰 영향을 받았는데, 0.5%(w/v) 농도의 NaCl 용액에서도 점성이 크게 떨어졌으며, 높은 온도에서는 점성이 비가역적으로 크게 증가하였다. Gel filtration 방법으로 조사한 EPSae의 분자량은 $2.5{\times}$10^6$ - $3.5{\times}$10^6$이었다. 냉동건조한 다당류를 전자현미경으로 관찰하였을 때, EPSbe는 섬유모양을 하였고, EPSae는 벌집모양의 망상구조를 하고 있었다.

• 한국식품과학회지
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• 제32권3호
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• pp.726-735
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• 2000

목질진흙버섯 자실체와 배양 균사체의 열수추출 및 열수 추출물의 에탄올, UF처리 분획들에 대한 약리 효과를 조사하여 다음과 같은 결과를 얻었다. Sarcoma 180 암세포 증식 저지율은 자실체 열수 총 분획(Fr. I)보다 비교적 고분자인 다당류 분획(Fr. II, Fr. III, Fr. IV)에서 $80.4{\sim}85.9%$로 높게 나타났다. 면역계에 미치는 영향으로 보체계 활성화는 $10.1{\sim}13.6%$, 대식세포의 NO 생성능은 자실체 및 균사체 모든 분획에서 생성되었으며, $TNF-{\alpha}$생성능은 NO 생성능이 높았던 분획들에서 높은 생성능을 나타내었다. 효소적 및 비효소적 지질과산화 유발에 대한 영향은 목질진흙버섯 자실체와 배양 균사체의 모든 분획에서 지질 과산화 억제능을 나타내었고, 유리기 제거능에 있어서는 농도의존적으로 효과가 있는 것으로 나타났으며, DPPH에 대한 Fr. III의 $SC_{50}$은 $32.0\;{\mu}g/mL$로 조사되었다. 상기와 같은 결과들로부터 목질진흙버섯 자실체와 배양 균사체의 열수추출 및 열수 추출물의 에탄올, UF처리 분획들은 항암활성, 면역활성, 지질 과산화 억제효과 및 유리기 제거능이 있음을 확인할 수 있었다.

• BMB Reports
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• 제40권5호
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• pp.636-643
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• 2007

This study was designed to identify and characterize the mechanism of macrophage activation by AAP, an acidic polysaccharide fraction isolated from the roots of Angelica sinensis (Oliv.) Diels. As a result, AAP significantly enhanced nitric oxide (NO) production and cellular lysosomal enzyme activity in murine peritoneal macrophages in vitro and in vivo. Furthermore, L-NAME, a specific inhibitor of inducible nitric oxide synthase (iNOS), effectively suppressed AAP-induced NO generation in macrophages, indicating that AAP stimulated macrophages to produce NO through the induction of iNOS gene expression and the result was further confirmed by the experiment of the increase of AAP-induced iNOS transcription in a dose-dependent manner. To further investigate, AAP was shown to strongly augment toll-like receptor 4 (TLR4) mRNA expression and the pretreatment of macrophages with anti-TLR4 antibody significantly blocked AAP-induced NO release and the increase of iNOS activity, and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) secretion.

• Asian-Australasian Journal of Animal Sciences
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• 제14권6호
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• pp.862-879
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• 2001

The walls of all higher plants are organized as a cellulosic, fibrillar phase embedded in a matrix phase composed of non-cellulosic polysaccharides, some proteins and, in most secondary walls, lignin. At the effective utilization of plant biomass, qualitative and quantitative analyses of plant cell walls are essential. Structural features of individual components are being clarified using newly developed equipments and techniques. However, "empirical" procedures to elucidate plant cell walls, which are not due to scientific definition of components, are still applied in some fields. These procedures may give misunderstanding for the effective utilization of plant biomass. In addition, interesting the investigation of wall organization is moving towards not only qualitatively characterisation, but also quantitation of the associations between wall components. These involve polysaccharide-polysaccharide and polysaccharide-lignin cross-links. Investigation of the associations is being done in order to understand the chemical structure, organization and biosynthesis of the cell wall and physiology of the plants. Procedures for qualitative and quantitative analyses based on the definition of cell wall components are reviewed focussing in nutritional elucidation of forage grasses by ruminant microorganisms.