• Title/Summary/Keyword: thin layer chromatography (TLC)

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Synthesis of 125I-labeled tetrazine for efficient radiolabeling of human serum albumin

  • Shim, Ha Eun;Jeon, Jongho
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.3 no.2
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    • pp.98-102
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    • 2017
  • We demonstrate a detail protocol for the radiosynthesis of a $^{125}I-labeled$ tetrazine prosthetic group and its application to the efficient radiolabeling of trans-cyclooctene-group conjugated human serum albumin (3) using inverse-electron-demand Diels-Alder reaction. Radioiodination of the stannylated precursor (2) was carried out by using [$^{125}I$]NaI and chloramine T as an oxidant at room temperature for 15 min. After HPLC purification of the crude product, the purified $^{125}I-labeled$ azide ([$^{125}I$]1) was obtained with high radiochemical yield ($65{\pm}8%$, n = 5) and excellent radiochemical purity (>99%). Inverse-electron-demand Diels-Alder reaction between ([$^{125}I$]1) and 3 gave the $^{125}I-labeled$ human serum albumin ([$^{125}I$]4) with more than 99% of radiochemical yield as determined by radio-thin-layer chromatography (radio-TLC). These results clearly indicate that the present radiolabeling method will be useful for the efficient and convenient radiolabeling of trans-cyclooctene-group containing biomolecules.

Enzymatic Production of High Molecular Weight Chitooligosaccharides Using Recombinant Chitosanase from Bacillus thuringiensis BMB171

  • Kang, Lixin;Jiang, Sijing;Ma, Lixin
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.45-50
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    • 2018
  • The chitosanase gene (btbchito) of Bacillus thuringiensis BMB171 was cloned and heterologously expressed in the yeast Pichia pastoris. After purification, about 300 mg of recombinant chitosanase was obtained from the 1-1 culture medium with a specific activity of 240 units/mg. Results determined by the combined use of thin layer chromatography (TLC) and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) showed that the chitooligosaccharides (COSs) obtained by chitosan (N-deacetylated by 70%, 80%, and 90%) hydrolysis by rBTBCHITO were comprised of oligomers, with degrees of polymerization (DP) mainly ranging from trimers to heptamers; high molecular weight chitopentaose, chitohexaose, and chitoheptaose were also produced. Hydrolysis products was also deduced using MS since the COSs (n) are complex oligosaccharides with various acetyl groups from one to two, so the non-acetyl COSs (GlcN)n and COSs with more acetyls (> 2) were not detected. The employment of this method in the production of high molecular weight COSs may be useful for various industrial and biological applications, and the activity of chitosanase has great significance in research and other applications.

강원도산 상엽과 누에의 혈당강하 활성물질의 특성

  • Kim, Sung-Hoon;Kim, Keun-Soo;Lee, Jin-Ha;Chung, Eul-Kwon;Park, Young-Shik;Park, You-Jeong;Lee, Hyeon-Yong
    • Microbiology and Biotechnology Letters
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    • v.25 no.4
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    • pp.391-395
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    • 1997
  • It was proved that both ethanol extracts from Mori Folium from Kangwon do and silk worm had higher inhibition acitivity on $\alpha$-glucohydrolase than the water extracts. In adding above 8.5 (mg/L) of silkworm extracts, the inhibition rate on $\alpha$-1,4 glucosidase was saturated while the inhibition rate was continuously increased in adding the extracts from Moli Folium. It was also found that the diethyl ether fraction showed much better inhibition activity than water fraction from ethanol extracts, yielding ca. 85% of inhibition rate for the extract of Moli Folium, compared to 91% for a commercially available hypoglycemic drug, Chloropropamide. In separating the diethyl ether fractions by Consecutive Sephadex gel filtration and Thin layer chromatography, three and four active spots were obtained from Moli Folium and silkworm, respectively. It is interesting that the similar Rf spots from both species among several spots in TLC have the highest inhibition acitivity on a target enzyme, which can imply that the active substances from both species are same or similar molecular weight and structure. Glucose-lowering activities of both speciese were also examined in vivo, showing that the fraction from Moli Folium had better activity than that from silkworm, and its activity was similar to that of a commercial drug.

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Anti-oxidative effects of Mori Folium, Mori Ramulus and Mori Cortex Radidus (상엽(桑葉), 상지(桑枝) 및 상백피(桑白皮)의 항산화 활성 연구)

  • Lim, Chi-Yeon;Lim, Se-Hyun;Cho, Su-In
    • Journal of Haehwa Medicine
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    • v.22 no.2
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    • pp.93-101
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    • 2014
  • Objective : This study was designed to investigate the anti-oxidative potentials to wide the usage of Morus alba L. derived from medicinal herbs on cosmoceutical fields. Methods : Anti-oxidative potentials were investigated by using several kinds of assays including electron donating ability, and the total contents of phenolic compounds were also measured. In some cases TLC (Thin Layer Chromatography) method were used to identify the patterns of phenolics in herbal extracts. Results : Aurones which are sub-types of flavonoids were observed in methanol extract of Mori Cortex Radidus, and total phenolic contents in Mori Cortex Radidus were more than in methanol extracts of Mori Folium and Mori Ramulus. Various kinds of phenolic compounds were observed in chloroform fraction of Mori Cortex Radidus. Conclusion : According to these results, the chloroform fraction of Mori Cortex Radidus could be the most possible resource that has potentials on anti-oxidant agents on cosmoceutical fields.

Studies on Changes of Aflatoxin Productivity and Properties by a Pseudomonad (녹농균과 진균류의 생물학적 상호작용에 의한 Aflatoxin 류의 생성능과 성질 변화에 대한 연구)

  • Lee, Young-Nam;Kim, Joo-Deuk
    • The Korean Journal of Mycology
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    • v.8 no.1
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    • pp.59-62
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    • 1980
  • Investigation on modification of aflatoxin structures by Pseudomonads was attempted as a biological detoxifiying process of mycotoxins. Firstly, when any variation of aflatoxin yield of Aspergillus parasiticus in a mixed culture with Pseudomonas aeruginosa was examined, there was no noticible effect by growth of Pseudomonads on aflatoxin yield of Asperillus sp. Secondly, when capacity of Pseudomonas aeruginosa utilizing aflatoxin as a carbon source for its growth was tested, there was some indication that aflatoxin might be used for growth of Pseudomonads. It was also noticed that the residual aflatoxin showed different migrating pattern compared with that of the intact aflatoxin by thin layer chromatography. Thirdly, the cell-free extract prepared from Pseudomonas aeruginosa grown in a glucose minimal medium supplemented with aflatoxin and the intact aflatoxins were incubated in the presence of $Mg^{++}$. After a certain length of incubation period, the reaction mixtures were applied on TLC plates. As a result, aflatoxins in the reaction mixture migrated differently as the control did. Such results may indicate that some changes of aflatoxin properties were induced by Pseudomonads.

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Survey and Control of the Occurrence of Mycotoxins from Postharvest Cereals (수확 후 곡류에 발생하는 진균독소의 캄색과 방제 1. 옥수수, 밀에서 분리한 Penicillium이 생산하는 주요 진균독소)

  • 오소영;정일민;백수봉;유승헌
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.700-704
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    • 1998
  • A total of 26 and 55 isolates of fungi were isolated from corn and wheat samples collected from different markets in Korea, respectively. The number of Penicillium isolates from corn and wheat was 9 and 33, respectively. The Penicillium species isolated from corn were P. chrysogenum (3 isolates) and P. oxalicum (6 isolates), and from wheat were P. aurantiogriseum (16 isolates), P. citrinum (1 isolate), P. commun (4 isolates), P. griseofulvum (1 isolate), P. verrucosum (7 isolates), and P. viridicatum (4 isolates). Production of major mycotoxins in the yeast extract sucrose medium cultures of Penicillium isolates was analysed. Penicillium cultures were extracted with chloroform and purified by thin-layer chromatograhy (TLC), and high performance liquid chromatography (HPLC). Among 9 isolates of Penicillium from corn, 2 isolates of P. chrysogenum produced patulin, 1 isolate of the fungus produced patulin and citrinin, 2 isolates of P. oxalicum produced penicillic acid, 4 isolates produced pencillic acid and griseofulvin. Of the 33 isolates of Penicillium from wheat, 6 isolates of P. aurantiogriseum produced patulin, 8 isolates produced penicillic acid, 1 isolate produced patulin and penicillic acid, 1 isolate of P. citrinum produced citrinin and patulin, 2 isolates of P. commun produced brefeldin A and patulin, 1 isolate of P. griseofulvum produced brefeldin A, griseofulvin and patulin. Five isolates of P. verrucosum produced patulin, 1 isolate of the fungus produced penicillic acid, and 3 isolates of P. viridicatium produced penicillic acid.

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Antioxidative Diarylheptanoids from the Fruits of Alpinia oxyphylla

  • Han, Jae-Taek;Lee, Sang-Yoon;Lee, Yonn-Hyung;Baek, Nam-In
    • Food Science and Biotechnology
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    • v.16 no.6
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    • pp.1060-1063
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    • 2007
  • The antioxidative activity of Alpinia oxyphylla was investigated through measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and inhibitory activity for linoleic acid peroxidation. Two antioxidative diarylheptanoids, yakuchinone A (1) and oxyphyllacinol (2), were isolated from the fruits of A. oxyphylla using thin layer chromatography (TLC) autographic assays. The DPPH scavenging activities of the compounds ($IC_{50}=1$, $57{\pm}2.1\;{\mu}M$; 2, $89{\pm}3.1\;{\mu}M$) were lower than vitamin C ($IC_{50}=51{\pm}1.1\;{\mu}M$), but higher than butylated hydroxytoluene (BHT, $IC_{50}=99{\pm}2.2\;{\mu}M$). Also, inhibitory activities for linoleic acid peroxidation of the compounds ($IC_{50}=1$, $0.19{\pm}0.011\;mM$; 2, $0.31{\pm}0.009\;mM$) were higher than those of vitamin C ($IC_{50}=0.59{\pm}0.017\;mM$) and BHT ($IC_{50}=0.52{\pm}0.014\;mM$). In addition the $^{13}C-NMR$ data of oxyphyllacinol (2) have been first reported in this paper.

Studies on the Separation of Maleic Anhydride Adducts of Soybean Oil (大豆油의 말레酸化生成物의 分別에 關한 硏究)

  • Suh Ho Park
    • Journal of the Korean Chemical Society
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    • v.21 no.1
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    • pp.67-72
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    • 1977
  • Author studied on the isolation technique and properties of the products obtained by the reaction of soybean oil with maleic anhydride. Before studying above purpose, oleic acid and linoleic acid contained in soybean oil were reacted with maleic anhydride. Thin-layer chromatography using silica coated plate and solvent system consisting of petroleum ether-diethyl ether = 7:3∼5:5 (v/v) resulted in good effects on the isolation of maleic anhydride adducts. Four fractions were separated and fraction No. 2 were analyzed by IR & UV spectrophotometer, obtaining the following conclusions. In the case of oleic acid adduct and linoleic acid adduct, it were conjectured that the succinic acid type adduct was formed. In the case of conjugated $C_{18}$ fatty acid adduct, cyclohexene type adduct was formed which differed from oleic acid and linoleic acid adduct.

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Functional Mechanism of Plant Growth Retardation by Bacillus subtilis IJ-31 and Its Allelochemicals

  • Kim, Won-Chan;Rhee, In-Koo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1375-1380
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    • 2012
  • We previously isolated a rhizobacterium (Bacillus subtilis IJ-31) and demonstrated that its associated allelochemicals could indicate plant growth retardation. However, little is known about how the growth of plants is regulated by B. subtilis IJ-31 and its allelochemicals. In this study, we investigated whether plant growth retardation in this relationship occurred through the inhibition of gibberellin (GA) biosynthesis. GA $3{\beta}$-hydroxylase activity was found to be inhibited by B. subtilis IJ-31 and hydrocinnamic acid (HCA), which is one of the allelochemicals produced by B. subtilis IJ-31. Additionally, thin layer chromatography (TLC) demonstrated that B. subtilis IJ-31 culture broth and HCA both inhibit GA $3{\beta}$-hydroxylase (MBP-GA4) activity. The retardation of plants by HCA was then confirmed in vivo and in vitro using a Ryegrass and Arabidopsis growth retardation assay. Furthermore, treatment with either B. subtilis IJ-31 culture extract or its allelochemicals resulted in significant down-regulation of XTR9 gene expression in Arabidopsis. Overall, we identified the functional mechanism of plant growth retardation by B. subtilis IJ-31 and its allelochemicals.

Synthesis of 125I-labeled thiol-reactive prosthetic group for site-specific radiolabeling of human serum albumin

  • Shim, Ha Eun;Song, Lee;Jeon, Jongho
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.4 no.2
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    • pp.85-89
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    • 2018
  • We demonstrate a detail protocol for the radiosynthesis of an $^{125}I$-labeled MSTP prosthetic group and its application to the efficient radiolabeling of human serum albumin (HSA). Radioiodination of the precursor (2) was carried out by using $[^{125}I]$NaI and chloramine T as an oxidant at room temperature for 15 min. After HPLC purification of the crude product, the purified $^{125}I$-labeled MSTP ($[^{125}I]1$) was obtained with high radiochemical yield ($73{\pm}5%$, n = 3) and excellent radiochemical purity (>99%). Site-specific reaction between ($[^{125}I]1$) and HSA gave the $^{125}I$-labeled human serum albumin ($[^{125}I]3$) with more than 99% of radiochemical yield as determined by radio-thin-layer chromatography (radio-TLC). These results clearly demonstrate that the present radiolabeling method will be useful for the efficient and convenient radiolabeling of thiol-bearing biomolecules.