• Title/Summary/Keyword: Medium pressure liquid chromatography

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Antifungal Substances from Streptomyces sp. A3265 Antagonistic to Plant Pathogenic Fungi

  • Nguyen, Van Minh;Woo, E-Eum;Kim, Ji-Yul;Kim, Dae-Won;Hwang, Byung Soon;Lee, Yoon-Ju;Lee, In-Kyoung;Yun, Bong-Sik
    • Mycobiology
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    • v.43 no.3
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    • pp.333-338
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    • 2015
  • In a previous study, we identified a Streptomyces sp., A3265, as exhibiting potent antifungal activity against various plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, and Rhizoctonia solani. This strain also exhibited a biocontrolling effect against ginseng root rot and damping-off disease, common diseases of ginseng and other crops. In this study, we isolated two antifungal substances responsible for this biocontrolling effect via Diaion HP-20 and Sephadex LH-20 column chromatography, medium pressure liquid chromatography, and high-performance liquid chromatography. These compounds were identified as guanidylfungin A and methyl guanidylfungin A by spectroscopic methods. These compounds exhibited potent antimicrobial activity against various plant pathogenic fungi as well as against bacteria.

Anti-inflammatory and Anti-oxidative Activities of Flavonoids Extracted from Dendranthema indicum Flowers in Jeju Island (제주 자생 감국 꽃 추출물 유래 flavonoid 화합물의 항산화 및 항염 활성)

  • Hyun, Ju Mi;Jo, Yeon Jeong;Kim, Yun Beom;Park, Sung-Min;Yoon, Kyung-Sup;Lee, Nam Ho
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1259-1267
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    • 2019
  • Anti-inflammatory and anti-oxidative activities were examined on the extract of Dendranthema indicum (D. indicum) flowers. The flowers were extracted two times for 24 h each with 70% ethanol. Upon the biological activities screening, the ethanol extract exhibited potent free radical scavenging activities and inhibited the production of nitric oxide on LPS-induced RAW264.7 macrophages effectively without causing cell toxicity. Further purification by medium pressure liquid hromatography (MPLC) and identification of the isolates led to identification of cynaroside (1) and apigetrin (2). The chemical structures of the isolated compounds were elucidated based on spectroscopic data including nuclear magnetic resonance (NMR) spectra, as well as comparison of the data to the literature values. Also, the quantitative analysis of the compounds was perfromed by high-performance liquid chromatography (HPLC). The isolates 1 and 2 were determined to inhibite the nitric oxide (NO) production dose-dependently. Based on these results, it was suggested that D. indicum extract could be useful as anti-inflammatory agents in cosmetics applications.

Three Phenolic Glycosides from Gastrodia elata

  • Chae, Sung-Wook;Lee, A-Yeong;Lee, Hye-Won;Yoon, Tae-Sook;Moon, Byeong-Cheol;Choo, Byung-Kil;Kim, Ho-Kyoung
    • Journal of Applied Biological Chemistry
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    • v.51 no.2
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    • pp.61-65
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    • 2008
  • Three phenolic glycosides, gastrodin (1), parishin (2), and parishin B (3) were isolated from the n-BuOH fraction of Gastrodia elata (Orchidaseae) by medium pressure liquid chromatography. Their chemical structures were identified by the interpretation of their spectral data and direct comparison with literature values. Gastrodin and parishin were isolated as the major constituents in G. elata. Gastrodin may be a suitable index component for quality control of G. elata.

Complete 1H-NMR and 13C-NMR spectral assignment of five malonyl ginsenosides from the fresh flower buds of Panax ginseng

  • Wang, Yu-Shuai;Jin, Yin-Ping;Gao, Wei;Xiao, Sheng-Yuan;Zhang, Yu-Wei;Zheng, Pei-He;Wang, Jia;Liu, Jun-Xia;Sun, Cheng-He;Wang, Ying-Ping
    • Journal of Ginseng Research
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    • v.40 no.3
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    • pp.245-250
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    • 2016
  • Background: Ginsenosides are the major effective ingredients responsible for the pharmacological effects of ginseng. Malonyl ginsenosides are natural ginsenosides that contain a malonyl group attached to a glucose unit of the corresponding neutral ginsenosides. Methods: Medium-pressure liquid chromatography and semipreparative high-performance liquid chromatography were used to isolate purified compounds and their structures determined by extensive one-dimensional- and two-dimensional nuclear magnetic resonance (NMR) experiments. Results: A new saponin, namely malonyl-ginsenoside Re, was isolated from the fresh flower buds of Panax ginseng, along with malonyl-ginsenosides Rb1, Rb2, Rc, Rd. Some assignments for previously published $^1H$- and $^{13}C$-NMR spectra were found to be inaccurate. Conclusion: This study reports the complete NMR assignment of malonyl-ginsenoside Re, $Rb_1$, $Rb_2$, Rc, and Rd for the first time.

Development of Doxorubicin overproducing Streptomyces Strain using Protoplast Regeneration (방선균 원형질체 재생에 의한 독소루비신 고생산성 균주개발)

  • 박희섭;박현주;김용훈;임상민;김동일;류욱상;김상린;김응수
    • KSBB Journal
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    • v.18 no.4
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    • pp.289-293
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    • 2003
  • Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. In order to increase doxurubicin productivity, a semi-industrial doxorubicin-producing Streptomyces strain named BR-Dox was cultured in a R2YE liquid medium containing CaCO$_3$, and then converted to a cell wall-free protoplast using lysozyme treatment method, followed by PEG-mediated cell wall regeneration. Among several protoplast-regenerated Streptomyces BR-Dox strains, two independent isolates named BR-Dox4 and BR-Dox6 were visually selected using thin layer chromatography (TLC) based on the pigment overproducing phenotype. Comparing with Streptomyces BR-Dox parental strain, two protoplast-regenerated strains, BR-Dox4 and BR-Dox6 exhibited 25.2% and 12.2% higher doxorubicin productivity analyzed by high pressure liquid chromatography (HPLC), respectively. This result suggests that a protoplast-regeneration of an antibiotics-producing Streptomyces strain should be a promising strain development approach for antibiotics overproduction in Streptomyces species.

Biological activity of flavonoids from Sonchus brachyotus

  • Lee, Jeong Min;Yim, Mi-Jin;Kim, Hyun-Soo;Ko, Seok-Chun;Kim, Ji-Yul;Shin, Jung Min;Lee, Dae-Sung
    • Fisheries and Aquatic Sciences
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    • v.24 no.12
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    • pp.428-436
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    • 2021
  • The aim of this study was to isolate and identify secondary metabolites from Sonchus brachyotus and evaluate their antioxidant and anti-inflammatory activities. In this study, we isolated three flavonoids from a 70% EtOH extract by Medium Pressure Liquid Chromatography (MPLC) and prep-High-Performance Liquid Chromatography (HPLC). To evaluate the biological activities (antioxidant and anti-inflammatory) of these flavonoids, their in vitro inhibitory activities against lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) generation, nitric oxide (NO) production, and prostaglandin E2 (PGE2) secretion were determined. We successfully identified three flavonoids, namely luteolin (1), luteolin-7-O-β-D-glucoside (2), and luteolin-7-O-β-D-glucuronide (3) by spectral analyses. Luteolin (1) at 20 ㎍/mL inhibited ROS generation, NO production, and PGE2 secretion by 48.6%, 61.28% and 12.10%, respectively, and luteolin-7-O-β-D-glucoside (2) inhibited NO and PGE2 generation by 67.03% and 20.82%, respectively. Luteolin (1) and luteolin-7-O-β-D-glucoside (2) showed similar anti-inflammatory activities; however, luteolin (1) was observed to be a stronger antioxidant. Besides antioxidant and anti-inflammatory activities, S. brachyotus extract containing luteolin (1) and luteolin-7-O-β-D-glucoside (2) is considered to possess diverse biological activities. The results indicate that S. brachyotus is an edible medicinal plant, which is believed to be significant resource of functional foods.

Identification of Polyphenol Substances (MP-1) from Seagrass, Phyllospadix Japonica Makino (잘피 게바다말로부터 폴리페놀 물질(MP-1)의 분리 및 동정)

  • Kim, Hae-Seon;Park, Nyun-Ho;Suk, Ho-young;You, Sang-guan;Woo, Jung-Hee
    • Korean Journal of Environmental Agriculture
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    • v.41 no.1
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    • pp.50-54
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    • 2022
  • BACKGROUND: Phyllospadix japonica Makino is a perennial plant belonging to the family Zosteraceae. This species is native to the eastern coast of the Korean Peninsula, and it is found attached to rocks on the seashore. As with all seagrass species, P. japonica is also known to play a major role in protecting the marine environment, and studies on its physiological activities suggestive of its antioxidant, antibacterial and anti-inflammatory potentials have been reported. In this study, purification and structural analysis were performed to identify the polyphenol substances derived from P. japonica. METHODS AND RESULTS: An polyphenol substance MP-1 was purified from the 70% aqueous methanol extract of P. japonica using Diaion® HP-20 column chromatography, ODS column chromatography, and medium-pressure liquid chromatography (MPLC). The purified MP-1 was identified as rosmarinic acid having a molecular weight of 360 and a molecular formula of C18H16O8 through electrospray ionization (ESI)-mass and nuclear magnetic resonance (NMR) spectroscopic analysis. CONCLUSION(S): This study highlights the processes used for the identification of the polyphenol substance derived from P. japonica. Rosmarinic acid, the polyphenol derived from P. japonica identified by this study, is a kind of bioactive substance mainly present in plants. These findings provide an important starting point and are valuable for future studies on bioactive substances in seagrass.

Development of High Cordycepin-Producing Cordyceps militaris Strains

  • Kang, Naru;Lee, Hyun-Hee;Park, Inmyoung;Seo, Young-Su
    • Mycobiology
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    • v.45 no.1
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    • pp.31-38
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    • 2017
  • Cordyceps militaris, known as Dong-Chong-Xia-Cao, produces the most cordycepin among Cordyceps species and can be cultured artificially. For these reasons, C. militaris is widely used as herb or functional food in the East Asia. In this study, we developed a new strain of C. militaris that produces higher cordycepin content than parent strains through mating-based sexual reproduction. Twenty parent strains were collected and identified as C. militaris based on internal trasncrived spacer and rDNA sequences. Seven single spores of MAT 1-1 idiomorph and five single spores of MAT 1-2 idiomorph were isolated from 12 parent strains. When 35 combinations were mated on the brown rice medium with the isolated single spores, eight combinations formed a stroma with a normal perithecia and confirmed mated strains. High pressure liquid chromatography analysis showed that mated strain KSP8 produced the most cordycepin in all the media among all the tested strains. This result showed due to genetic recombination occurring during the sexual reproduction of C. militaris. The development of C. militaris strain with increased cordycepin content by this approach can help not only to generate new C. militaris strains, but also to contribute to the health food or medicine industry.

Content Analysis of Rutin in the Leaves of Boehmeria nivea Harvested in Different Regions of South Korea by HPLC-UV

  • Cho, Sunghun;Quilantang, Norman G.;Lee, Ju Sung;Kim, Young-Mi;Kim, Ho Bang;Cho, Eun Ju;Jung, Yong-Su;Lee, Sanghyun
    • Natural Product Sciences
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    • v.24 no.1
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    • pp.36-39
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    • 2018
  • Phytochemical analysis of Boehmeria nivea (Bn) leaves by medium pressure liquid chromatography led to the isolation of a flavonoid glycoside identified by spectroscopic analysis as rutin. The amount of rutin in the leaves of Bn harvested from nine regions in South Korea (Bn 1-9) which were collected on the months of June, July, August, and September was determined by HPLC-UV analysis. A gradient elution program that utilizes a $Discovery^{(R)}$ C18 ($4.6{\times}250mm$, $5{\mu}m$) column and mobile phase composed of 1% acetic acid-water: acetonitrile (90:10 to 60:40 for min) was followed. The injection volume and flow rate were $10{\mu}l$ and 1 mL/ min, respectively. UV detection was set at 350 nm. Results show that Bn-8 harvested in September reported the highest content of rutin among the samples analyzed. This study provides a basis for the optimal harvest time of Bn which maximizes the yield of rutin.

Production, Structural Elucidation, and In Vitro Antitumor Activity of Trehalose Lipid Biosurfactant from Nocardia farcinica Strain

  • Christova, Nelly;Lang, Siegmund;Wray, Victor;Kaloyanov, Kaloyan;Konstantinov, Spiro;Stoineva, Ivanka
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.439-447
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    • 2015
  • The objective of this study was to isolate and identify the chemical structure of a biosurfactant produced by Nocardia farcinica strain BN26 isolated from soil, and evaluate its in vitro antitumor activity on a panel of human cancer cell lines. Strain BN26 was found to produce glycolipid biosurfactant on n-hexadecane as the sole carbon source. The biosurfactant was purified using medium-pressure liquid chromatography and characterized as trehalose lipid tetraester (THL) by nuclear magnetic resonance spectroscopy and mass spectrometry. Subsequently, the cytotoxic effects of THL on cancer cell lines BV-173, KE-37 (SKW-3), HL-60, HL-60/DOX, and JMSU-1 were evaluated by MTT assay. It was shown that THL exerted concentration-dependent antiproliferative activity against the human tumor cell lines and mediated cell death by the induction of partial oligonucleosomal DNA fragmentation. These findings suggest that THL could be of potential to apply in biomedicine as a therapeutic agent.