• Title/Summary/Keyword: Periodate oxidation

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Purification and Characterization of Complement-activating Acidic Polysaccharides from the Fruits of Capsicum annuum

  • Paik, Soon-Young;Ra, Kyung-Soo;Chang, In-Seop;Park, Yoon-Chang;Park, Hee-Sung;Baik, Hyung-Suk;Yun, Jong-Won;Choi, Jang-Won
    • BMB Reports
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    • v.36 no.2
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    • pp.230-236
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    • 2003
  • Hot water-soluble crude polysaccharide (HCAP-0) that was obtained from the fruits of Capsicum annuum showed potent anti-complementary activity. The activity was unchanged by pronase digestion, but decreased by periodate oxidation. The HCAP-0 was fractionated by DEAE ion-exchange chromatography to give two major fractions, HCAP-II and III. These two fractions were finally purified by gel filtration to give HCAP-IIa, HCAP-IIIa1, and IIIa2 fractions that had high anti-complementary activities. The HCAP-IIIa1 and IIIa2 consisted of homogeneous polysaccharides. The anti-complementary activities were unaffected by treatment with polymyxin B, indicating that the modes of complement activation were not due to preexisting lipopolysaccharide. The molecular weight and sugar content of HCAP-IIIa2 had potent anti-complementary activity. The highest yields were 55 kDa and 75.9%, and the molar ratio of galactose (Ara:Gal, 1.0:4.6) was higher than other sugars. The crossed immuno-electrophoresis showed that both classical and alternative pathways were activated by HCAP-IIIa2.

Purification and Characterization of Complement System Activating Polysaccharide from the Bark of Kalopanax pictus N. (음나무 수피로부터 보체계 활성화 다당의 정제 및 특성)

  • Shin, Keum;Ra, Kyung-Soo;Paik, Ki-Hyon
    • Journal of the Korean Wood Science and Technology
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    • v.20 no.4
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    • pp.73-84
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    • 1992
  • It was observed that the hot-water extract of the bark of Kalopanax pictus N. had the highest anti-complementary activity among the 11 kinds of forest materials. Methanol-and ethanol-soluble portions had low anti-complementary activities, but crude polysaccharide. HKP-0 had a high activity of 80%. HKP-0 contained 54.8% of total sugar and 27.9% of protein. The neutral sugars of HKP-0 consisted of mainly arabinose, galactose and glucose. HKP-4 fraction obtained by cetavlon treatment of HKP-0 showed the highest anti-complementary activity of 90%. The activity was not changed by pronase digestion bu decreased greatly by periodate oxidation. HKP-4 consisted of mainly arabinose and glucose with molar ratio of 1.0 : 22.4, HKP-4-I, an unabsorbed fraction from HKP-4 on DEAE Sepharose CL-6B column showed higher yield and activity than those of absorbed fractions. HKP-4-I was homogeneous, and its molecular weight was about 25,000. HKP-4-I contained 84.0% of neutral sugar and consisted of arabinose and glucose with molar ratio of 1.0 : 11.2. The anti-complementary activity of HKP-4-I was not decreased by the treatment of polymyxin B, and the polysaccharide activated both classical and alternative pathway in complement system. Void volume fraction obtained from HKP-4-I hydrolyzed with ${\alpha}$-amylase on Sephadex G-25 column only had a high anti-complementary activity.

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Chemical Analysis of Acidic Proteo-heteroglycans with Anti-complementary Activity from the Hot-Water Extract of Fomitella fraxinea (장수버섯 자실체로부터 분리한 항보체 활성 단백다당체의 화학적 분석)

  • Yoon, Sang-Hong
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.502-510
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    • 1998
  • The hot-water extract of fruiting bodies in Fomitella fraxinea had potent anti-complementary activities. After fractionation of water-soluble polysaccharides by DEAE-Sephadex A-25 column chromatography, major anti-complementary activity was concentrated into the FF-AP1 among three polysaccharides (FF-NP, FF-AP1, FF-AP2). FF-AP1 was fractionated into $FF-AP1{\alpha}$ and $FF-AP1{\beta}$ obtained from the adsorbed fraction and unadsorbed fraction by affinity chromatography using a ConA-sepharose 4B column, respectively. $FF-AP1{\beta}$, which exihibited the highest anti-complementary activities had an IR absorption peak of $890cm^{-1}$, and a M.W. of about 15,000 (gel filtration). Anti-complementary activity of FF-AP1 decreased greatly by pronase treatment and periodate oxidation. $FF-AP1{\beta}$ responsible for potent anti-complemenary activities of Fomitella fraxinea was an acidic protein-containing heteroglycan consisted of 48% glucose, 13% mannose, and 12% galactose as major component sugars, 9.6% protein, 6% uronic acids.

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Characterization of polysaccharide A-1 from Opuntia ficus-indica and it's protection effect on alcoholic induced hepatic oxidative stress (Opuntia ficus-indica 다당 A-1의 특성 및 알코올유도 간 산화스트레스의 보호 효과)

  • Ryu, Il-Hwan;Kwon, Ji-Wung;Lee, Eoh-Jin;Yun, Young-Gab;Kwon, Tae-Oh
    • Herbal Formula Science
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    • v.17 no.2
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    • pp.163-174
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    • 2009
  • Reactive oxygen species(ROS) can induce hepatotoxicity and trigger apoptosis in the liver. In this study, we investigated the sulfated polysaccharide A-1 from Opuntia ficus-indica against alcoholic oxidative stress in human liver Hep G2 cell. An antioxidant substance A-1 obtained from the enzymatic extract of Opuntia ficus-indica fruit was purified by DEAE-cellulose ion exchange and sephadex G-100 gel permeation chromatography. The purification yield and molecular weight were 14.3% and 1.8 KDa, respectively. The A-1 predominately contained arabinose, galactose, rhamnose and also sulfate group. The structure of A-1 was investigated by periodate oxidation, FT-IR spectroscopy, $^1H$-NMR spectroscopy. The A-1 mainly composed of alternating unit of ${\rightarrow}4$)-$\alpha$-L- Rapp-2-$SO_3^-$-$\alpha$-L-Galp-($1{\rightarrow}$ and branched linkage of $\beta$-D-Arbp- ($5{\rightarrow}$. The antioxidative activity was measured using the SOD, CAT activity and GSH assay, respectively. The expression of Nrf2 protein was analyzed by western blotting. The viable cell count analyzed by autofluorescence. Oxidative stress induced by ethanol(1 M) were dramatically reduced by A-1 treatment. A-1 also prevented cell death induced by oxidative stress. It also increased expression Nrf2 protein level. We concluded that sulfated polysaccharide A-1 from Opuntia ficus-indica effectively protect Hep G2 liver cell from alcoholic oxidative stress.

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An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid (판토텐산의 분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Park, Youn-Sick;Bae, Gun-Won
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1009-1014
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    • 2000
  • In order to detect pantothenic acid (PA), conditions for enzyme-linked immunosorbent assay (ELISA) were established. Anti-PA-BSA antibody was produced from rabbits immunized with PA-bovine serum albumin (BSA) conjugates which were prepared by the bromoacetyl chloride [Bc] method (PA-BSA[Bc]) and by the periodate oxidation [Po] method (PA-BSA[Po]). PA-BSA[Bc] and PA-BSA[Po] was used as a coating antigen for competitive indirect(ci)ELISA. The Anti-PA-BSA[Po] antibody on ciELISA showed no competitive reaction. The detection limit of PA by ciELISA using Anti-PA-BSA[Bc] antibody was 1 ppm. The Anti-PA-BSA[Bc] antibody showed little cross-reactivity to PA derivatives such as pantoyllactone, pantetheine, pantothenyl alcohol, and acetyl CoA. The detection limit of PA by microbiological assay (MBA) was 10 ppb. Assay recoveries of PA in egg, cow's liver, and lettuce by ciELISA were 109, 64, and 344%, respectively, comparing with the MBA results.

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Physicochemical Properties and Gel Forming Properties of Mungbean and Buckwheat Crude Starches (녹두와 메밀 조전분의 이화학적 특성 및 겔 형성)

  • 주난영;이혜수
    • Korean journal of food and cookery science
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    • v.5 no.2
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    • pp.1-8
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    • 1989
  • The physicochemical properties and gel forming properties of mungbean and buckwheat crude starches were investigated. The results were as follows; 1. The granule size and shape of mungbean crude starch were $11~32\mu\textrm{m}$ and oval, and those of buckwheat crude starch were $3~10 \mu\textrm{m}$ and polygonal. 2. The amylose conteut of mungbean crude starch and buckwheat crude starch were 78.0% and 26.4% respectively. 3. The blue value of mungbean crude starch and buckwheat crude starch were 1.030 and 0.409, respectively. 4. Periodate oxidation of mungbean crude starch resulted that amylose had the average molecular weight of 95, 648, degree of polymerization of 590 and amylopectin had the degree of branching of 5.4, glucose unit per segment of 18.6, and periodate oxidation of buckwheat crude starch resulted that amylose had the average molecular weight of 133, 690, degree of polymerization of 825 and amylopectin had the degree of branching of 5.2, glucose unit per segment of 19.2 5. Water binding capacity of mungbean crude starch was 185.1% and that of buckwheat was 209.9% 6. The pattern of change in swelling power of mungbean crude starch for increasing temperature started to increase at $60^{\circ}C$ and increased rapidly from $70^{\circ}C$, and that of buckwheat increased slowly from $60^{\circ}C$ to $90^{\circ}C$ without rapid increase. 7 The ranges of gelatinization temp. of mungbean and buckwheat crude starches were 63. 9-$109^{\circ}C$ and 52.5-84.$2^{\circ}C$, respectively. 8. The gelatinization patterns for 6% munbean crude starch and 8% buckwheat crude starch were investigated by Brabender amglograph. Mungbean crude starch showed the initial pasting temperature of 77.6$^{\circ}C$ without peak height, and buckwheat crude starch showed that of $62.5^{\circ}C$ without peak height. In addition, sensory evaluation for sample starch gels (mungbean, buckwheat, cowpea) was done. 1. The difference of sensory characteristics for each starch gel was significant. 2. The sample starch gels were regarded as 'Mook' by pannels. 3. 74.44% of the degree of Mooklike was explained by hardness.

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Structural Analysis of the Antitumor Active Exo-polysaccharide Produced by Submerged Cultivation of Ganoderma lucidum Mycelium (영지(Ganoderma lucidum) 균사체의 액체배양에 의한 세포외 항암활성 다당류의 구조분석)

  • Lee, Shin-Young;Kang, Tae-Su
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.76-81
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    • 1999
  • Exo-polysaccharide obtained from the submerged cultivation of Ganoderma lucidum mycelium was fractionated. The structural analysis of the acidic exo-polysaccharide fraction (BWS-DA-GI), showing high antitumor activity, was carried out and compared to the mycelial acidic fraction (MWS-DA-GI). The major sugar constituents of the fraction of BWS-DA-GI were glucose, galactose and mannose in the molar ratio of 2.5 : 2.1 : 2.5. The minor components in this fraction were xylose and fucose. While the major sugar constituents of the mycelial acidic fraction of MWS-DA-GI were galactose, fucose, mannose and glucose. The trace components in this fraction was xylose. From the results of periodate oxidation, Smith degradation, affinity chromatography and methylation analysis, the chemical structures of the two fractions, BWS-DA-GI and MWS-DA-GI were both determined as ${\beta}-1,3$ glucans. It was also estimated that BWS-DA-GI had a $1{\rightarrow}6$ glucosidic linkage and MWS DA-GI had $1{\rightarrow}4$ and $1{\rightarrow}6$ glucosidic linkages. The molecular weights of these fractions, MWS-DA-GI and MWSDA-GI were estimated as $1.2{\times}10^6\;and\;1.0{\times}10^6$ dalton, respectively.

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Physicochemical Properties of Cowpea Crude and Refined Starch (동부 조전분 및 정제전분의 이화학적 특성)

  • 윤혜현;이혜수
    • Korean journal of food and cookery science
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    • v.3 no.1
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    • pp.31-36
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    • 1987
  • The purpose of this study is to investigate the physicochemcal Properties of the cowpea crude and refined starch and to present the basic data for physicochemical factor which gives the properties of Mook to cowpea starch gel. Water binding capacity of crude starch was 235. In and that of refined starch was 186.0%. The pattern of change in swelling power and solubility for increasing temperature started to increase at $60^{\circ}C$ and increased rapidly from $70^{\circ}C$, for both of crude and refined starch. The optical transmittance of 0.2% crude and refined starch suspensions were increased from $65^{\circ}C$ and showed rapid increasement during 68~$80^{\circ}C$, and their curves showed two-stage processes. The gelatinization pattern for 6n crude and refined starch suspensions were investigated by the Brabender amylograph. The corves showed the pasting temperature of $72.0^{\circ}C$ and $72.1^{\circ}C$, peak height of 11303.U. ($88.0^{\circ}C$) and 970 B.U. ($83.5^{\circ}C$) for crude and refined starch, respectively, and both showed high viscosities when cooling. Blue values for crude and refined starch were 0.369 and 0.376 respectively. Alkali number of crude and refined starch were 7.77 and 7.34, and reducing values were 3.60 and 2. 10, respectively. Amylose content of cowpea starch was 33.7%. Periodate oxidation of the starch fractions resulted that amylose had the average molecular weight of 23590, degree of polymerization of 146 and amylopectin had the degree of branching of 3.42, glucose unit per segment of 29.

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Screening of Complement-System Activating Polysaccharide from Edible Plants and Its Action Mode (식물성 식품재료로부터 보체계 활성화 다당의 검색 및 그 활성검토)

  • Shin, Kwang-Soon;Ra, Kyung-Soo;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.25 no.3
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    • pp.197-203
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    • 1993
  • Screenings were performed on edible plants to examine their complement-system activating ability (anti-complementary activity) by hemolytic complement assay $(TCH_{50})$. Among 38 kinds of plant extracts, 5 kinds showed relatively strong anti-complementary activity which decreased $TCH_{50}$ more than 60% comparison with control and the order of activity was Zingiber officinale>Colocasia antiquorum>Capsella bursapastoris>Ginkgo biloba>Alium monanthum in $1000{\mu}g/ml$. The anti-complementary activity of ZR-1 prepared from the root of Zingiber officinale which was showed the most potent activity, did not change by pronase treatment, but decreased greatly by periodate oxidation. These results indicate that not protein moiety but carbohydrate moiety in ZR-1 fraction may also contribute to the anti-complementary activity. Also, the anti-complementary activity of ZR-1 was reduced partially in the absence of the $Ca^{2+}$ ion. When crossed immunoelectrophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the ZR-1 in $Ca^{2+}$ free condition, a cleavage of C3 precipitin line was observed. Furthermore this polysaccharide fraction considerably inhibited $ACH_{50}$. These results also indicate that the mode of complement activation by polysaccharide from Zingiber officinale is via not only the classical pathway but also the alternative pathway.

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Isolation and Characterization of Complement System Activating Polysaccharides from the Hot Water Extract of the Leaves of Capsicum annuum L. (고추잎 열수추출물로부터 보체계 활성화 다당의 분리 및 특성)

  • 이용세;나경수;정상철;서형주;박희성;백형석;최장원
    • Journal of Life Science
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    • v.12 no.1
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    • pp.87-95
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    • 2002
  • It was observed that the hot water extract of the leaves of Capsicum annuum L., a Korean edible plant, had a potent anti-complementary activity. Crude polysaccharide fraction(CAL-0) was obtained by methanol reflux, ethanol precipitation, dialysis and lyophilization. CAL-0 contained 51.8% of total sugar, 8.2% of uronic acid and 16.8% of protein, and consisted of mainly arabinose, galactose and glucose as neutral sugars and galacturonic acid as uronic acid. The anti-complementary activity of CAL-0 decreased greatly by periodate oxidation, but was not changed by pronase treatment. Also, the anti-complementary activity of CAL-0 was reduced partially in the absence of the $Ca^{2+}$ ion. The crude polysaccharide CAL-0 was found to activate the C3 component both in the presence and in the absence of $Ca^{2+}$ through the crossed-immunoelectrophoresis suggesting that those involved in both classical and alternative complement pathway CAL-0 was further separated to an unabsorbed fraction(CAL-1) and six absorbed fractions(CAL-2longrightarrowCAL-7) on DEAE Sepharose CL-6B ion exchange column. Among them four major fractions in activity and yield were obtained, and consisted mainly of arabinose, galactose and glucose with various molar ratios. The major fraction, CAL-2, was purified to give a high molecular fraction(CAL-2-I) and a low molecular fraction(CAL-2-II) on Sepharose CL-6B column. The anti-complementary activity of CAL-2-I, a molecular weight of about 61,000, was higher than it of CAL-2-II.-II.