• Title/Summary/Keyword: Sephadex LH-20 column chromatography

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Inhibitory Effect of Cacao Bean Husk Extract on Glucosyltransferase from Streptococus mutans B13 (Cacao Bean Husk 추출물의 Glucosyltransferase 저해효과)

  • 권익부;이용우안봉전이신영
    • KSBB Journal
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    • v.8 no.1
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    • pp.75-82
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    • 1993
  • The inhibitory effect of cacao bean husk (CBH) extract on glucosyltransferase(GTasc) from Streptococcus mutans B13 was investigated. Water solube extract from CBH showeda sarong inhibitory effect (88-89%) on GTase from Streptococcus mutans Bl3. GTase inhibitors from sequential extraction by hot water or water-methanol had the strongest inhibition. Sources, fermentation, and types of solvents and fumigation processes did not influence the effect. These active compounds proved to be polyphones through acid hydrolytic analysis of the precipitates by ammonium sulfate or ethanol and proteinase K. It was also confirmed by additional column chromatography of Sephadex G-50, Sephadex LH-20 and DEAE-Sephdex A-50.

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Brassinosteroids in Rice Bran and Polished Rice (미강과 백미의 Brassinosteroid 활성물질)

  • Park, Kyoung-Won;Park, Jong-Dae;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.360-365
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    • 1996
  • To investigate the presence of brassinosteroids in rice bran and pㅐlished rice, they were extracted with MeOH. The extracts were purified through sequential procedure of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 column chromatography and charcoal adsorption chromatography. The activity of brassinosteroids was monitored by the rice inclination test and its presence was confirmed in each purification step. The purified active components were further separated by TLC and HPLC. Brassinosteroids in active fractions of rice bran were identified as castasterone and teasterone by HPLC. We acknowledge that our work is probably the first report of brassinosteroids in mature seeds of rice The more amount of brassinosteroids was confirmed in rice bran than polished rice. The contents of castasterone and teasterone which were identified in rice bran were 0.15 ng/g and 0.37 ng/g, respectively.

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Identification of 3-methoxy-4-hydroxybenzoic acid and 4-hydroxybenzoic acid with Antioxidative and Antimicrobial Activity from arachis hypogaea Shell (땅콩껍질에서 항균 및 항산화활성이 있는 3-methoxy-4-hydroxybenzoic acid와 4-hydroxybenzoic acid의 동정)

  • 위지향;박근형
    • KSBB Journal
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    • v.15 no.5
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    • pp.464-468
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    • 2000
  • The methanol extract of Arachis hypogaea shell showed antioxidative and antimicrobial activity. The methanol extract was successively purified by solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography and octadecylsilane column chromatography. The purified active substances were isolated by high performance liquid chromatography, and were identified as 3-methoxy-4-hydroxybenzoic acid and 4-hydroxybenzoic acid by LC-MS and GC-MS. The amount of 3-methoxy-4-hydroxybenzoic acid and 4-hydroxybenzoic acid were 3.8mg and 9.8 mg per kg of shell, respectively.

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Isolation and Characterization of 4-hydroxycinnamic Acid with Antimicrobial Activity from Aralia eiata (두릅에서 항미생물활성을 갖는 4-hydroxycinnamic acid의 분리 및 동정)

  • Ma, Seung-Jin;Kuk, Ju-Hee;Ko, Byoung-Seob;Park, Keun-Hyung
    • Applied Biological Chemistry
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    • v.39 no.4
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    • pp.265-267
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    • 1996
  • The methanol extracts of AraIia elata showed antimicrobial activity against bacteria. The antimicrobial active principle was successively purified with solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, silica gel partition column chromatography and HPLC. The active substance was isolated by HPLC on $C_{18}$ column with an acetic acid-MeOH system. The active substance was identified as trans-4-hydroxycinnamic acid by MS, $^1H-NMR\;and\;^{13}C-NMR$.

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지초뿌리 유래의 기능성 물질의 탐색

  • 문영환;조정용;위지향;문제학;박근형
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.134.2-135
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    • 2003
  • 지초(Lithospermum efthrorhizon)뿌리는 예로부터 한방약제로 이용되어져 왔으며, 지초 뿌리에 함유된 naphtoquinon계 색소물질인 shikonin은 항암, 항균, 항바이러스, 항염증 등의 효과가 있다고 보고되었다. 그러나 지초뿌리에 함유된 기능성 물질에 관한 연구는 미비한 실정이다. 이에 본 연구에서는 지초뿌리에서 기능성 물질의 탐색 및 기능성 해명연구를 위하여 지초뿌리를 ethanol로 추출한 후 이 추출물을 n-hexane, EtOAc, MeOH로 순차적으로 추출하여 MeOH추출물을 얻었다. 이 MeOH추출물을 silica gel adsorption column chromatography, ODS column chromatography, Sephadex LH-20 column chromatography로 순차 정제한 후 Shodex Asahipak column을 이용한 GPC-HPLC에 의해 활성 물질을 분리하였다. 분리된 물질들은 1H-NMR, 13C-NMR, COSY, HSQC, HMBC, FAB-MS 등의 기기분석을 통해 당 관련 화합물인 것으로 판명되었다.

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Purification of ACE Inhibitor from Soybean Paste (된장으로부터 Angiotensin Converting Enzyme 활성 저해물질의 정제)

  • Suh, Hyung-Joo;Suh, Dae-Bang;Chung, Soo-Hyun;Whang, Jong-Hyun;Sung, Ha-Jin;Yang, Han-Chul
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.441-446
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    • 1994
  • Seven commercial soybean paste were tested for ACE inhibition effect. In purification of ACE inhibitor from No. 2 soybean paste, acetone fraction $(50{\sim}80%)$ had 57% protein yield with 92.8% ACE inhibition effect. Inhibitor was purified from acetone fraction of soybean paste by Sephadex G-25, Sephadex LH-20 and ODS column chromatography and HPLC. $IC_{50}$ value of the purified inhibitor was 0.6 mg/ml. The inhibitor showed the competitive inhibition patterns on ACE. Amino acid analysis showed that the peptides consist of Ala, Phe, Leu, Glu, Gly, Ser, and Asp.

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Purification and Characteristics of Tyrosinase Inhibitor Produced by Actinomycetes F-97 (방선균 F-97이 생산하는 Tyrosinase 저해제의 정제 및 특징)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Kim, Jin-O;Yi, Dong-Heui
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.153-158
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    • 2008
  • An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 ($NH_4^+$ type) column chromatography, silica gel column chromatography, $C_{18}$ column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The ${\lambda}_{max}$ value of this inhibitor in water was 194nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, cone. $H_2SO_4$, and $KMnO_4$ tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of $100^{\circ}C$ for 50 minutes and pH $4{\sim}9$. The $IC_{50}$ value of this inhibitor was $19.2{\mu}g/ml$ for mushroom tyrosinase. In $1,000{\mu}g/ml$ inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.

Chemical Structures of the Compounds Isolated from the Edible Mushroom Sarcodon aspratus (능이버섯(Sarcodon aspratus)으로부터 분리한 저분자 화합물의 화학구조)

  • Kang, Hee-Chol;Yun, Bong-Sik;Yu, Seung-Hun;Yoo, Ick-Dog
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.298-302
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    • 2000
  • In our investigation on chemical constituents of edible mushrooms, seven compounds I-VII have been isolated from the methanolic extract of the fruit body of Sarcodon aspratus. The methanolic extract was separated by solvent partition, silica gel column chromatography and Sephadex LH-20 column chromatography, and compounds I-VII were finally purified by preparative HPLC or TLC. Their structures were assigned as 4-hydroxybenzoic acid methyl ester, 4-hydroxybenzaldehyde, cyclo(Ala-Pro), adenosine, nicotinamide, BL V, linoleic acid, respectively, on the basis of mainly $^1H\;NMR$ spectra.

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Inhibitory Effect of Hexane Fraction from Myagropsis myagroides on Pancreatic α-Amylase In Vitro

  • Pak, Won-Min;Kim, Koth-Bong-Woo Ri;Kim, Min-ji;Cho, Ji-Young;Ahn, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.328-333
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    • 2015
  • A Myagropsis myagroides (Mm) methanol extract showed α-amylase inhibitory activity of 13% at a concentration of 5 mg/ml. Results showed that the hexane fraction from the Mm methanol extract exhibited α-amylase inhibitory activity with an IC50 value of 4.24 mg/ml. The hexane fraction was separated using silica-gel column chromatography, and six subfractions were obtained. The fraction eluted with CHCl3:MeOH = 50:1 showed the highest α-amylase inhibitory activity with an IC50 value of 0.72 mg/ml. This fraction was purified using Sephadex LH-20 column chromatography and an octadecyl silica (ODS) Sepak cartridge, obtaining seven subfractions. Fraction (Fr.) 4 also showed a strong α-amylase inhibitory activity with an IC50 value of 0.75 mg/ml. Fr. 4 was purified by Sephadex LH-20 column chromatography and ODS Sepak cartridge, obtaining six subfractions. Fr. 4-2 was identified as sargachromanol I with an IC50 value of 0.40 mg/ml, and the inhibition pattern analyzed from Lineweaver-Burk plots revealed it to be an uncompetitive inhibitor. These results suggest that Mm has potential as a natural antidiabetes agent.

Phenolic Compounds from Acer tegmentosum Bark (산겨릅나무 수피의 페놀성 화합물)

  • Kwon, Dong-Joo;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.145-151
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    • 2007
  • To investigate the chemical constituents of Acer tegmentosum, the bark were collected, air-dried and extracted with 70% aqueous acetone. Then it was successively partitioned with n-hexane, $CH_2Cl_2$, EtOAc and $H_2O$. Repeated Sephadex LH-20 column chromatography on the EtOAc soluble fraction gave five phenolic compounds. Their structures were elucidated as (+)-catechin (1), (-)-epi-catechin (2), Q-epicatechin-3-O-gallate (3), gallic acid (4) and 6'-0-galloylsalidroside (5) on the basis of spectroscopic evidences using $^1H-NMR$, $^{13}C-NMR$, 2D-NMR and MS spectroscopy, (-)-epicatechin-3-Ogallate (3), gallic acid (4), 6'-Ogalloylsalidroside (5) have not been reported in this plant yet.