• Title/Summary/Keyword: TLC analysis

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Diversity, Saccharification Capacity, and Toxigenicity Analyses of Fungal Isolates in Nuruk (누룩곰팡이 분리균의 다양성 및 당화능 분석과 독소생산능 조사)

  • Kim, Min Sik;Kim, Sinil;Ha, Byeong-Seok;Park, Hye-Young;BaeK, Seong-Yeol;Yeo, Soo-Hwan;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.191-200
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    • 2014
  • Nuruk samples collected from various regions in Korea were investigated in terms of fungal contents and diversity. In measurement of colony forming unit (CFU) in Nuruk suspensions on DRBC agar, Nuruk samples MS4, MS8, and MS10 were among the highest fungal density, with $1,278.9{\pm}21.6$ (${\times}10^4$), $1,868.0{\pm}27.7$ (${\times}10^4$), and $775.1{\pm}19.2$ (${\times}10^4$) were among the samples showing the highest fungal density. CFU per 20 mg Nuruk, respectively. The majority of fungal components were yeasts, including Pichia anomala, P. kudriavzevii, Kluyveromyces marxianus, and Saccharomycopsis fibuligera, whereas Aspergillus oryzae and Rhizopus oryzae, the representative Nuruk fungi, were predominant only in the low fungal density Nuruks (MS2, MS5, and MS11). Saccharification capability of the fungal isolates was assessed by measurement of amylase activity in the culture broth. The highest amylase activity was found in A. niger and A. luchuensis, followed by S. fibuligera. A. oryzae and R. oryzae showed fair amylase activity but significantly lower than those of the three fungal species. R. oryzae was suggested to play an additional role in degradation of ${\beta}$-glucan in crop component of Nuruk since R. oryzae was the only fungus that showed ${\beta}$-glucanase activity among the fungal isolates. To confirm the safety of Nuruk, aflatoxigenicity of the isolated Aspergillus was estimated using the DNA markers norB-cypA, aflR, and omtA. All of the isolates turned out to be non-aflatoxigenic as evidenced by the deletion of gene markers, norB-cypA and aflR, and the absence of aflatoxin in the culture supernatants shown by TLC analysis.

Antioxidant Activity and Component Analysis of Fermented Lavandula angustifolia Extracts (라벤더 발효추출물의 항산화 활성과 성분 분석)

  • Park, Soo-Nam;Ahn, You-Jin;Won, Bo-Ryoung;Kang, Myung-Kyu;Kim, Jai-Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.2
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    • pp.125-134
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    • 2009
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component of non-fermented and fermented Lavandula angustifolia extracts were investigated. The ethyl acetate fraction of fermented extract (5.95 ${\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of L. angustifolia extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract (1.45 ${\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of L. angustifolia on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The L. angustifolia extracts suppressed photohemolysis in a concentration dependent manner (1 ${\sim}$ 50 ${\mu}g/mL$). The inhibitory effect of L. angustifolia extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase were determined with ethyl acetate fraction of L. angustifolia extract (144.80 ${\mu}g/mL$) and ethyl acetate fraction of fermented extract (122.40 ${\mu}g/mL$). Fractions of ethyl acetate and fermented extracts showed both 3 band in TLC and 3 peaks, 2 peaks in HPLC (340 nm), respectively. In each chromatography, fractions of ethyl acetate both from non-fermented and fermented L. angusfifolia have rosmarinic acid in common. These results indicate that the component and content of non-fermented and fermented extracts of L. angustifolia are different. Both of the extract of L. angustifolia can be used as an antioxidant.

Antioxidative Activities and Antiaging Effects of Geranium nepalense Extracts (현초 추출물의 항산화 활성 및 항노화에 관한 연구)

  • Lee, Keun-Ha;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.1
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    • pp.25-35
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    • 2008
  • In this study, the antioxidative effects and inhibitory effects on elastase and tyrosinase of Geranium nepalense extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of Geranium nepalense were in the order: 50% ethanol extract(15.0 ${\mu}g/mL$)${\mu}g/mL$). ${\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities($OSC_{50}$) of some Geranium nepalense extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescense assay. The order of ROS scavenging activities were 50% ethanol extract($OSC_{50},\;0.23{\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Deglycosylated flavonoid fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Geranium nepalense on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Geranium nepalense extracts suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated flavonoid fraction exhibited the most prominent celluar protective effect (${\tau}_{50}$, 676.7 min at 50 ${\mu}g/mL$). The inhibitory effect of aglycone fraction on tyrosinase($IC_{50}$, 70.0 ${\mu}g/mL$) and elastase ($IC_{50}$, 19.9 ${\mu}g/mL$) was very high. Aglycone fractions obtained from the deglycosylation reaction of ethyl-acetate fraction among the Geranium nepalense extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (370 nm). Two components were identified as quercetin(composition ratio, 15.3%), kaempferol(82.8%). These results indicate that extract/fractions of Geranium nepalense can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Geranium nepalense extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

Method for Supplementing Lecithin to Ginseng Extract (레시틴이 강화된 인삼 추출물 제조 방법)

  • Park, Soon-Hye;Kim, Il-Woong;Kim, Dong-Man;Kim, Si-Kwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.9
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    • pp.1245-1250
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    • 2006
  • This study was carried out to develop the method of preparing lecithin-fortified ginseng extract. Firstly, soybean lecithin was mixed with soybean oil (LCS) in varying ratio (2.5%, 5%, 10% and 20%). Then, one part volume of LCS was mixed with three parts volume of ginseng extract with 10% solid matter content and the mixture was vortexed vigorously. Finally, the mixture was spinned at the speed of 3,000 rpm for 30 minutes to separate oil and aqueous ginseng extract layer (AG). AG was then subjected to qualitative and quantitative analysis of phospholipids and ginsenosides. Fatty acid composition and crude fat content before and after LCS was determined. Stability of lecithin in ginseng extract was determined by analyzing phospholipid content in the one third upper and lower layer of the concentrated AG in Falcon tubes while storing the LCS treated concentrated AG in 4, 25 and 40oC for 6 months. Ratio of lecithin transferred to AG increased with the increase in lecithin content of soybean oil. There was no significant change in fatty acid composition and crude fat content, and ginsenoside content in the ginseng extract before and after LCS treatment. TLC and HPLC pattern of saponin fraction before and after treating the ginseng extract with LCS demonstrated no observable difference. There was no change in lecithin content in the upper and lower one third layer of ginseng extract in the tubes after storing the concentrated AG in 4, 25 and $40^{\circ}C$ for 6 months. Ginsenosides HPLC pattern was not changed when stored the LCS-treated ginseng extract in those conditions for six months, indicating satisfiable stability of the LCS-treated concentrated ginseng extract. From these results, it can be concluded that treatment of the ginseng extract with lecithin containing soybean oil is a labor effective method with satisfiable stability to fortify lecithins to ginseng extract.

Cellular Protective Effect and Component Analysis of Euphorbia humifusa Extracts (땅빈대 추출물의 세포 보호 효과 및 성분 분석에 관한 연구)

  • Kim, Sun-Young;Won, Doo-Hyun;Lim, Myoung-Sun;Park, Soo-Nam
    • Korean Journal of Pharmacognosy
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    • v.41 no.4
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    • pp.264-269
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    • 2010
  • In this study, the cellular protective effect, antioxidative property and component analysis of Euphorbia humifusa extracts were investigated. The ethyl acetate fraction ($3.68\;{\mu}g/mL$) and aglycone fraction ($3.15\;{\mu}g/mL$) of Euphorbia humifusa extract showed prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of Euphorbia humifusa extract on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction ($0.43\;{\mu}g/mL$) and aglycone fraction ($0.35\;{\mu}g/mL$) of extract showed higher ROS scavenging activity than L-ascorbic acid ($1.50\;{\mu}g/mL$). The cellular protective effects of fractions of Euphorbia humifusa extract on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction and aglycone fraction of extract protected cellular membranes against ROS in a concentration dependent manner ($5{\sim}25\;{\mu}g/mL$), and was more effective than (+)-${\alpha}$-tocopherol, lipid peroxidation chain blocker. Aglycone fraction from Euphorbia humifusa extract showed 2 bands in TLC and 2 peaks in HPLC. In HPLC chromatogram of aglycone fraction, peak 1 and peak 2 were identified as quercetin and kaempferol, respectively. And these components are very effective as antioxidant. Thus, these results indicate that fractions of Euphorbia humifusa extracts can function as antioxidant in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Fractions of Euphorbia humifusa extracts can be applicable to new functional cosmetics for antioxidant.

A Study on Internal-External Morphology and Pattern Analysis in Angelicae Pubescentis Radix (독활(獨活)의 외부(外部) 및 내부형태(內部形態)와 이화학패턴연구)

  • Kim, Hong-Jun;Kim, Ja-Young;Choi, Go-Ya;Jeong, Seung-Il;Ju, Young-Sung
    • Korean Journal of Oriental Medicine
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    • v.12 no.3 s.18
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    • pp.101-115
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    • 2006
  • The purpose of this study was to introduce the differential standard of the types of Angelicae Pubescentis Radix. We established the classificatory standard according to the external and internal morphology and the physicochemistrical pattern for the four types of Aucklandiae Radix. The results follow: 1. At the external shape, Angelica pubescens f. biserrata has rising lenticel and dark brown surface, Its section is light gray and its xylem is yellow gray. Aralia continentalis has big stem mark in upper portion, its section has no dense tissue with yellow gray. Heracleum hemsleyanum has dark brown surface and little lenticel, its cortical layer is white yellow and xylem is isabella and powdered. 2. At the internal shape, Angelica pubescens f. biserrata has phloem with half of the root and rare vessel. Aralia continentalis has phloem with two-thirds of the root and it has broad cutting and putting vessel that formed 1-2 row. Heracleum hemsleyanum also has phloem with half of the root and wide scattered latex tube and many large vessel. 3. At the TLC pattern, Heracleum hemsleyanum has remarkable dark spot at $R_{f}$ 0.23 on the sulphuric acid color pattern test, but others have faint. 4. At the HPLC pattern, all samples have generally patterns. But Angelica pubescens f. biserrata shows diminutive continentalic acid content and the peak at Rt 20.278min comes out on Heracleum hemsleyanum, but do not come out on Aralia continentalis and Angelica pubescens f. biserrata. Heracleum hemsleyanum has remarkable peak at Rt 20.278min, but shows no peak at Rt 29.023min unlike Aralia continentalis or Angelica pubescens f. biserrata. Also Aralia continentalis and Angelica pubescens f. biserrata show one remarkable peak at Rt 29.023, but Heracleum hemsleyanum do not show. Consequently, Aralia continentalis and Angelica pubescens f. biserrata are comparable whit continentalic acid content and Heracleum hemsleyanum is comparable with the peak at Rt 20.273 and Rt 29.023. So it is thought that content of continentalic acid and the peaks at Rt 20.278 and Rt 29.023 can apply to differentiate a species from other. It is considered the results of this study will be furnished the basis to succeeding studies and it is needed to extensive comparative study for the same genus-degree of relatedness.

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Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.

Characterization of Nitrile-hydrolyzing Enzymes Produced from Rhodococcus erythropolis (니트릴 분해효소 생산균인 Rhodococcus erythropolis의 발굴 및 효소 특성 연구)

  • Park Hyo-Jung;Park Ha-Joo;Uhm Ki-Nam;Kim Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.204-210
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    • 2006
  • Ethyl (S)-4-chloro-3-hydroxybutyrate is a useful intermediate for the synthesis of Atorvastatin, a chiral drug to hypercholesterolemia. In this research, two 4-chloro-3-hydroxybutyro-nitrile-degrading strains were isolated from soil sample. They were identified as Rhodococcus erythropolis strains by 16S rRNA analysis. The nitrile-degrading enzyme(s) were suggested to be nitrile hydratase and amidase rather than nitrilase from the result of thin layer chromatography analysis. The corresponding genes were obtained by PCR cloning method. The predicted protein sequences had identities more than 96% with nitrile hydratase ${\alpha}-subunit$, nitrile hydratase ${\beta}-subunit$, and amidase of R. erythropolis. The 4-chloro-3-hydroxybutyronitrile-hydrolyzing activities in both strains were increased dramatically by ${\varepsilon}-caprolactam$ which was known as good inducer for nitrile hydratase. Both intact cells and cell-free extract could hydrolyze the nitrile compound. So, the intact cell and the enzymes could be used as potential biocatalyst for the production of 4-chloro-3-hydroxybutyric acid.

Toxicity and Characteristics of Antifungal Substances Produced by Bacillus amyloliquefaciens IUB158-03 (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 생화학적 특성 및 독성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1672-1678
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    • 2009
  • The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

Isolation of Alliin in Garlic and Its Quantitative Determination by High Performance Liquid Chromatography and Studies on the Antimicrobial Efforts of Alliin and Ethanol Extracts from Korean Garlic(Alliium sativum L.) (마늘 중 고속 액체 크로마토그래피에 의한 알린의 분리 및 정량과 Alliin과 에탄올 추출물의 항균효과에 관한 연구)

  • 위성언
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.296-302
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    • 2003
  • First. the purification and analysis of alliin in garlic from different origins by alliin-HPLC determination method were studied. Allinase in garlic was inactivated by heating in boiling water followed by extraction of alliin in garlic with 80% methanol. To remove free amino acids and alliin homologs in garlic, garlic extract was separated by cation exchange column which was packed with amberlite CG-120 resin using 40L d-water as eluent. Alliin in garlic extract was crystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=70:29:1 and then recrystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=75:24:1. Obtained alliin was identified by melting point. TLC, microscope observation and mass spectrometry. High performance liquid chromatography (HPLC) following pre-column derivatization of cystein derivatives with o-phthaldialdehyde/2-mercaptoethanol has succeessfully been applied to the analysis of various garlics. Each alliic of standard solution and garlic extract was derivatized to isoindole derivative by o-phthaldialdehyde /2-mercaptoethanol and then analyzed by HPLC. Six point calibration was done by using alliin peak area. Lineality was observed at 0 ∼ 1.0mg/ml of alliin concentration. Weighted regression line function was Y=6254X - 256077. By this function, alliin contents in various garlics were 0.34 ∼ 0.73% fresh weight. Second study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogenes (Eubacterium limonsum, Bacteroides fragilis, Salmonella typhimurium, Salmonella typhi, Shigella sonnei, Kiebsiella pneumoniae, Enterobacter cloacae, Pserdomonas aeruginosa, Escherichia coli). For antimicrobial effects against microorganism, totally minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000ppm. MIC of ethanol extract were 1,250 to 10,000ppm.