• Title/Summary/Keyword: 구조동정

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Purification and characterization of antifungal compounds produced by Bacillus subtilis KS1 (Bacillus subtilis KS1이 생산하는 항진균물질의 정제 및 특성)

  • Ryoo, Sung-Woo;Maeng, Hack-Young;Maeng, Pil-Jae
    • The Korean Journal of Mycology
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    • v.24 no.4 s.79
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    • pp.293-304
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    • 1996
  • A bacterial strain, KSl, possessing strong antifungal activity was isolated from soil samples of ginseng fields and identified as Bacillus subtilis. In greenhouse test, the culture filtrate of B. subtilis KS1 showed strong protective effect against several fungal diseases of agricultural plants such as cucumber gray mold and wheat leaf rust. In addition, the crude butanol fraction of the culture filtrate exhibited antagonistic effect against several fungi including plant or human pathogens, such as Botrytis maydis, Chytridium lagenarium and Candida albicans. The antifungal compound, SW1, produced by B. subtilis KS1 was purified through consecutive chromatographic separations on a pep-RPC column and a ${\mu}$ Bondapak $C_{18}$ reverse phase column. Temperature and pH showed little effect on the stability of the compound in the ranges $-20-121^{\circ}C$ and pH 4.0-10.0, respectively. The composition and structural characteristics of SW1 were analysed by HPLC and by $^1H-,\;^1H-^1H-COSY$, NOESY, COSY-NOESY and HOHAHA NMR spectroscopy, respectively, which revealed that the compound belongs to iturin A, a typical cyclic antifungal compound produced by B. subtilis. In contrast to the previously reported iturin A compounds which have one or no $-CH_3$ side chain in the hydrophobic hydrocarbon chain of ${\beta}-amino$ acids, SW1 was shown to have a ${\beta}-amino$ acid containing 12-carbon skeleton with two $-CH_3$ side chains.

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화학독립영양미생물 Aeromonas sp. strain JS-1의 RubisCO 정제 및 특성

  • Na, Suk-Hyeon;Bae, Sang-Ok;Kim, Min-Jeong;Kim, Seong-Jun;Jeong, Seon-Yong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.461-464
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    • 2002
  • A Chemoautotroph identified as an Aeromonas sp. strain JS-1 was isolated from fresh water. Aeromonas sp. strain JS-1 used the $H_2$ and $CO_2$ as energy and carbon sources, respectively. Growth characteristics for improving the $CO_2$ fixation rate were examined in batch cultivation. Its results shown that the optimal growth appeared at culture conditions of $35^{\circ}C$, pH 7 and NaCl 0.1%(w/v). Some hydrogen-oxidizing bacteria were reported that the enzyme activity of ribulose 1,5-bisphosphate carboxylase- oxygenase (RubisCO-EC 4.1.1.39), in the key enzyme of the Calvin-Benson cycle. A RubisCO was purified from a chemoautotrophic bacterium, Aeromonas sp. strain JS-1. the enzyme was purified by ammonium sulfate precipitation, DEAE-sepharose CL-6B and gel filtration chromatography. The RubisCO showed that molecular mass was about 560KDa from gel filtration chromatography and nondenaturing PAGE, and the RubisCO was confirmed to consist of $L_8S_8$ enzyme structure by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A large subunit was about 56KDa and small one was about 15kDa. The Km values of the enzyme for ribulose 1,5-bisphosphate(RUBP), $NaH^{14}CO_3$, and $Mg^{++}$ were estimated to be 0.25mM, 5.2mM, and 0.91mM, respectively. The optimum temperature for RubisCO enzymatic activity were $50^{\circ}C$, and the enzymatic activity was stable up to $45^{\circ}C$.

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Gene Expression Pattern during Early Embryogenesis and Transcriptional Activities of Estrogen Receptor-Related Receptor(ERR) in Sea Urchin, Strongylocentrotus nudus (둥근성게(Strongylocentrotus nudus) Estrogen Receptor-Related Receptor(ERR)의 초기 발생시 유전자 발현 패턴과 전사 활성)

  • Maeng, Se-Jung;Kim, Mi-Soon;Sohn, Young-Chang
    • Development and Reproduction
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    • v.13 no.4
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    • pp.249-256
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    • 2009
  • The estrogen receptor-related receptors (ERRs) are a group of nuclear receptor superfamily of transcription factors. ERRs and estrogen receptors (ERs) have overlapping affinities for coactivators and DNA binding sites, but differ markedly in ligand binding and activation. The three mammalian ERR genes have been implicated in diverse physiological processes ranging from placental development to maintenance of bone density, whereas the molecular diversity, function, and regulation of ERRs in non-mammalian species are not well understood. In the present study, to investigate the involvement of ERR in transcription and embryogenesis in marine invertebrates, a cDNA encoding ERR (SnERR) was cloned from the gonad in Strongylocentrotus nudus, by polymerase chain reaction (PCR). The amino acid sequence of SnERR showed high homology with that of S. purpuratus (91%). A phylogenetic tree clearly showed that SnERR is a member of the ERR family and clustered in echinodermata group as supported by a high bootstrap value. We examined gene expression of SnERR during embryonic development of S. nudus using real-time PCR. During the embryonic development, the mRNA of ERR was significantly high levels in early development stages (4~64 cell) and larval stages. The SnERR slightly activated transcription through the classical estrogen response elements (EREs) in the presence of genistein. In addition, peroxisome proliferator-activated receptor $\gamma$ coactivator (PGC)-$1\alpha$ knwon as a coactivator of ERR enhanced the snERR-mediated transactivation, suggesting that the PGC-$1\alpha$ is a coactivator of SnERR.

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Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Kimchi (김치로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 규명)

  • Kim Uyo-Ju;Chang Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.196-203
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    • 2006
  • Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

Anti-wrinkle Compounds Isolated from the Seeds of Arctium lappa L. (우방자에서 분리한 주름개선 화합물)

  • Hwang, Ju-Young;Park, Tae-Soon;Kim, Dong-Hee;Hwang, Eun-Young;Lee, Jung-Noh;Lee, Ji-Young;Lee, Ghang-Tai;Lee, Kun-Kook;Son, Jun-Ho
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1092-1098
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    • 2012
  • This study was carried out to discover the skin wrinkle reducing components in the seeds of Arctium lappa. The isolation of a methylene chloride-soluble fraction of 70% ethanol extract from the seeds of Arctium lappa using a procollagen type-1 synthesis and MMP-1 activity resulted in the isolation and identification of four lignin compounds: arctiin, arctigenin, matairesinol, and diarctigenin. All structures were confirmed via NMR and MS spectroscopic data. To determine cell viability and procollagen type-1 synthesis, human dermal fibroblasts were treated with 10-100 ${\mu}M$. As a result, none of the four compounds showed cytotoxicity up to 50 ${\mu}M$. We also investigated their procollagen type-1 synthesis and MMP-1 inhibition activity and found that arctiin had the highest activity in terms of both procollagen synthesis and MMP-1 inhibition among all four compounds. Putting all the data together, we suggest that arctiin be used in cosmetics as an anti-wrinkle material.

Identification of Substrate Specificity Determinant of Achromobacter Protease I (API) and Catalytic Activity of Mutant D225E for Ornithine Substrate (Achromobacter Protease I (API)의 기질특이성 결정기의 동정과 변이체[D225E]의 Ornithine 기질에 대한 촉매활성)

  • Lim, Seong-Il;Kwon, Oh-Jin;Choi, Cheong
    • Applied Biological Chemistry
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    • v.40 no.3
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    • pp.189-195
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    • 1997
  • The structural basis of Iysine specificity of Achromobacter protease I (API) was investigated by means of site-directed mutagenesis. The precursor protein in which Glu190, one of the two candidates for determining Iysine specificity, was substituted by glutamine, aspartic acid or leucine was processed autocatalytically to attaln full pretense activity with lysine specificity. The substitution of the other candidate, Asp225, for asparagine or leucine produced no mature active forms of pro-API. The precursor protein of the mutant D225E slowly matured autocatalytically. The lysylendopeptidase activity of the mature D225E was 0.25% of that of native API, and this reduced activity is mainly due to a decrease in the affinity of the enzyme for lysine. These results suggest that Asp225 plays a critical rol in restricted substrate specificity as a lysylendopeptidase. However, D225E exhibited no measurable activity for synthetic ornithine substrate. Since the hydroxyl group of Ser194 in this mutant retained essentially the same reactivity to DFP or PMSF as that in native API, it can be noted that a methylene unit longer side chain of residue 225 is not compensated by a methylene unit shorter side chain at subsite P1 in the bound substrate.

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Isolation of Anticonvulsant Compounds from the Fruits of Schizandra chinensis$B_{AILI}$ (오미자(Schizandra chinensis $B_{AILI}$.) 열매로부터 항경련 활성물질의 분리)

  • Han, Jae-Taek;Ahn, Eun-Mi;Park, Jin-Kyu;Cho, Sung-Woo;Jeon, Seong-Gyu;Jang, Joong-Sik;Kim, Choong-Kwon;Choi, Soo-Young;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.43 no.1
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    • pp.72-77
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    • 2000
  • The repeated silica gel colum chromatographies of EtOAc fraction, showing anticonvulsant activity, obtained from MeOH extracts of Schizandra chinensis B. fruits led to isolation of a sesquiterpenoid, four lignans and a sterol glycoside. Their chemical structures were determined to be chamigrenal, gomisin A, gomisin H, gomisin N. schizandrin and daucosterol. Among them, schizandrin and daucosterol inhibited GABA degrative enzymes, succinic semialdehyde dehydrogenase and succinic semialdehyde reductase, respectively. It is postulated that the schizandrin and daucosterol are able to elevate the neurotransmitter GABA levels in central nervous system by inhibitory action on GABA degrative enzymes and act as anticonvulsant drugs.

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Studies on Antimicrobial and Antioxidative Activities of Extracts from Magnoliaceae (목련과 식물의 항균 및 항산화활성)

  • Lee, Sung-Suk;Lee, Hak-Ju
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.579-586
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    • 2010
  • This study was carried out to evaluate the antifungal and antioxidative activities of ethanol extracts from Magnoliaceae to investigate the possibility for the natural fungicides and food preservatives. The antifungal activities of ethanol extracts were evaluated as a hyphal growth inhibition rate using four plant pathogenic and five wood rot fungi. The high inhibition activity on the growth of fungi was shown in bark of Magnolia obovata that indicated more than 50% hyphal growth inhibition rate except Trametes versicolor, one of the white rot fungi. The antifungal activity was the highest in the ethanol extracts from M. obovata and the following was in order of M. kobus and M. sieboldii. The extract from bark in M. obovata showed higher antifungal activity than that from wood in the same species. Especially, the extracts from flower of M. denudata and M. liliflora indicated the high antifungal activities, while the other portions of same plants showed the low activities. On the other hand, a free radical scavenging method was adopted with 1,1-diphenyl-2-picrylhydrohydrazyl (DPPH) in order to test the antioxidative activities of ethanol extracts. The free radical scavenging activity was very high in the extracts from branch of Schizandra nigra and Kadsura japonica that showed more than 90% at the concentration of 100 ${\mu}g/m{\ell}$. It also turned out that the antioxidative activity of branch of S. nigra and K. japonica was similar to ${\alpha}$-tocopherol and butylated hydroxytoluene, one of the effective synthetic antioxidants. From these results, it can be suggested that the branches of S. nigra and K. japonica have the positive antioxidative activities and can be applied for the food preservatives and cosmetic ingredient.

Studies on Biological Activity of Wood Extractives (X VIII) -Isolation and Antioxidant Activity of Chemical Constituents from Maackia amurensis- (수목추출물의 생리활성에 관한 연구(X VIII) -다릅나무(Maackia amurensis) 수피의 추출성분의 분리 및 항산화 활성-)

  • Kim, Woo-Jin;Lee, Hak-Ju;Lee, Sang-Keug;Kang, Ha-Young;Choi, Don-Ha;Choi, Tae-Ho
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.135-144
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    • 2007
  • The dried barks of Maackia amurensis were ground, extracted with 95% EtOH, concentrated, and fractionated with a series of light petroleum ether, dichloromethane, ethyl acetate and water on a separatory funnel. Each fraction was concentrated, then a portion of dichloromethane and ethyl acetate soluble was chromatographed on a Sephadex LH-20 and silica gel 60 column using a various solvent system as eluents. The isolated compounds were identified by cellulose TLC, $^1H-$, $^{13}C-NMR$, COSY, NOESY, HMQC, HMBC, FAB and EI-MS. The structures were determined as: 7-O-$\beta$-D-glucopyranosyl-4'-methoxyisoflavone, 7-O-$\beta$-glucopyranosyl(1'''->6'')-$\beta$-D-glucopyranosyl-4'-methoxyisoflavone, 7-O-$\beta$-D-glucopyranosyl(1''''->6''')-$\beta$-D-glucopyranosyl(1'''->6'')-$\beta$-D-glucopyransoyl-4'-methoxyisoflavone, 7-O-$\beta$-D-glucopyranosyl-4', 6-dimethoxyisoflavone. The Free radical scavenging activity using DPPH of the isolated compounds were similar with that of BHT but lower than of $\alpha$-tocopherol.

Chemical and Mineralogical Characteristics of White Stones Excavated from Archaeological Sites (유적지 출토 백색 암석의 광물화학적 특성)

  • Han, Min-Su;Cho, Nam-Chul;Kim, Woo-Hyun;Choi, Kyung-Yong
    • Journal of the Mineralogical Society of Korea
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    • v.22 no.3
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    • pp.199-206
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    • 2009
  • The aim of this study is to investigate the relations among each white stone and their characteristics through the chemical and mineralogical analysis of them excavated from around some kilns in archaeological sites. Moreover it was intended to infer the function of kilns. Nine white stones were chosen from three different archaeological sites, the each name of which is Sanyang-ri, Youngdu-ri and Gwangdae-ri in Chungcheongnam-do. In the mineralogical characteristics of thin section, eight stones containing calcite are altered and recrystallized by metasomatism and hydrothermal process, and Calcites, major component minerals of limestone, were identified by the analysis of XRD. On the basis of this result, we can infer the stones to be classified into limestone groups. One the other hand, research revealed that one of the white stones was a white pelitic stone composed of quartz, microcline and muscovite. In the analysis of chemical composition, except for the white pelitic stone eight stones are mainly composed of CaO which is major component of Calcite. Besides, as a results of correlation analysis by using a chemical compositions of major and minor elements in white stones, little did each archaeological site have the relativity. In conclusion, each archaeological site was estimated that they would use a limestone ores extracted from different deposits, and there is a high possibility that the function of these kilns is to produce quicklime.