• Title/Summary/Keyword: Efficient Purification

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A Simplified Procedure for the Large-Scale Purification of Urokinase from Human Urine (인뇨로부터 유로키나제 대량정제공정의 단순화)

  • 정광회;선우명
    • KSBB Journal
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    • v.5 no.2
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    • pp.183-189
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    • 1990
  • An efficient method has been developed for the purification of urokinase from 1, 000 liter batches of human urine. The procedure involved precipitation of urokinase with 2mM zinc chloride, resuspension of the precipitate with 0.1M EDTA/0.5M Glycine solution, and CM-Toyopearl and benzamidine-Sepharose column chromatography. The purified urokinase was fully active and possessed a specific activity of 1.07$\times$105IU/mg. The recoveries ranged from 42 to 65% in several preparations(mean value was 51%). And the urokinase purified by this process consisted of about 13% of single chain urokinase (pro-urokinase) as evaluated by SDS-polyacrylamide gel electrophoresis in reducing condition and by S-2444 amldolytic activity under plasmin treatment.

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Refolding and Purification of Recombinant Human $Interferon-\gamma$ Expressed as Inclusion Bodies in Escherichia coli Using Size Exclusion Chromatography

  • Guan Yi-Xin;Pan Hai-Xue;Gao Yong-Gui;Yao Shan-Jing;Cho Man-Gi
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.122-127
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    • 2005
  • A size exclusion chromatography (SEC) process, in the presence of denaturant in the refolding buffer was developed to refold recombinant human $interferon-\gamma$ ($rhIFN-\gamma$) at a high concentration. The $rhlFN-\gamma$ was overexpressed in E. coli resulting in the formation of inactive inclusion bodies (IBs). The IBs were first solubilized in 8 M urea as the denaturant, and then the refolding process performed by decreasing the urea concentration on the SEC column to suppress protein aggregation. The effects of the urea concentration, protein loading mode and column height during the refolding step were investigated. The combination of the buffer-exchange effect of SEC and a moderate urea concentration in the refolding buffer resulted in an efficient route for producing correctly folded $rhIFN-\gamma$, with protein recovery of $67.1\%$ and specific activity up to $1.2\times10^7\;IU/mg$.

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.

Efficient Constitutive Expression of Cellulolytic Enzymes in Penicillium oxalicum for Improved Efficiency of Lignocellulose Degradation

  • Waghmare, Pankajkumar Ramdas;Waghmare, Pratima Pankajkumar;Gao, Liwei;Sun, Wan;Qin, Yuqi;Liu, Guodong;Qu, Yinbo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.740-746
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    • 2021
  • Efficient cellulolytic enzyme production is important for the development of lignocellulose-degrading enzyme mixtures. However, purification of cellulases from their native hosts is time- and labor-consuming. In this study, a constitutive expression system was developed in Penicillium oxalicum for the secreted production of proteins. Using a constitutive polyubiquitin gene promoter and cultivating with glucose as the sole carbon source, nine cellulolytic enzymes of different origins with relatively high purity were produced within 48 h. When supplemented to a commercial cellulase preparation, cellobiohydrolase I from P. funiculosum and cellobiohydrolase II from Talaromyces verruculosus showed remarkable enhancing effects on the hydrolysis of steam-exploded corn stover. Additionally, a synergistic effect was observed for these two cellobiohydrolases during the hydrolysis. Taken together, the constitutive expression system provides a convenient tool for the production of cellulolytic enzymes, which is expected to be useful in the development of highly efficient lignocellulose-degrading enzyme mixtures.

Effect of surfactant on the micelle process for the pre-purification of paclitaxel (Paclitaxel 전처리를 위한 마이셀 공정에서의 계면활성제 영향)

  • Jeon, Keum-Young;Kim, Jin-Hyun
    • KSBB Journal
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    • v.23 no.6
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    • pp.557-560
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    • 2008
  • The micelle process was developed for pre-purifying paclitaxel from plant cell cultures of Taxus chinensis, giving a high purity and yield. The approach in this work was to transfer paclitaxel in the crude extract to an aqueous surfactant solution as a micelle, allowing organic solvents to be used for removal of lipids and non-polar impurities. In this work, the effects of various surfactants such as CPC, CTMAC, LTMAC, SDS, AOT, Tween, PEG, and Triton were examined on the yield, purity, and phase separation time in micelle process. Among these surfactants, CTMAC (5%, w/v) gave the best result in terms of paclitaxel yield (${\sim}99%$), purity (${\sim}21%$), and phase separation time (30 min). The use of micelles in the pre-purification process allows for rapid and efficient separation of paclitaxel from interfering compounds and dramatically increases the yield and purity of crude paclitaxel for subsequent purification steps.

A Study on Efficient Improvement Method of Rainwater Utilization Facilities in Jeju Island (제주지역 빗물이용시설의 효율적 개선방안 연구)

  • Park, Won-Bae;Moon, Deok-Cheol;Koh, Gi Won
    • Journal of Soil and Groundwater Environment
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    • v.17 no.6
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    • pp.1-8
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    • 2012
  • This study is to suggest a few efficient ways of rainwater utilization, through monitoring and analyzing 143 rainwater storage systems and 110 artificial recharge systems, which are installed in the recommended facilities by law, among the rainwater harvesting systems in Jeju Island. In the case that catchment facilities are damaged, rainwater could be contaminated by leaves and debris so that the rates of rainwater usages come to be lower. It is possible that contaminated rainwater could contaminate artificial recharge wells or rainwater discharging out of the rainwater harvesting system could result in flood and damage for the downgradient area. For maintaining high quality of rainwater and increasing rainwater utilization rate, it is necessary to install screening facilities and purification plant functioning precipitation and filtration. Also, in order to efficiently preclude the overflowing rainwater exceeding storage capacity, it is recommended to associate rainwater storage tanks with artificial recharge well or infiltration trench facilities.

A Study on the Development Trend of Explosion and Combustion Energy (폭발.연소 에너지의 개발 방향에 관한 연구)

  • Shin, Chang-Yong;Ahn, Myung-Seog;Jo, Myung-Chan
    • Explosives and Blasting
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    • v.27 no.2
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    • pp.56-60
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    • 2009
  • In view of physics, energy is defined as the ability to work. The use of natural gas and nuclear power have been increased since 1980s to replace fossil fuels such as coal and petroleum. Recently, solar energy, wind power, tidal power, and geothermal energy have been considered as promising alternative energy sources to overcome environmental pollution. However, their energy efficiencies are much lower than those of chemical energies such as nuclear power, explosive, and petroleum gas. In this study, the present situation of the green energy was reviewed to seek out the way to overcome the limit of the environmental (alternative) energy. Also, purification, application and development trend of the highly efficient alternative energy sources were investigated.

Low-temperature Synthesis of Graphene-CdLa2S4 Nanocomposite as Efficient Visible-light-active Photocatalysts

  • Zhu, Lei;Oh, Won-Chun
    • Journal of the Korean Ceramic Society
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    • v.52 no.3
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    • pp.173-179
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    • 2015
  • We report the facile synthesis of graphene-$CdLa_2S_4$ composite through a facile solvothermal method at low temperature. The as-prepared products were characterized by X-ray diffraction (XRD) and by Scanning electron microscopy (SEM) with energy dispersive X-ray (EDX) analysis and BET analysis, revealing the uniform covering of the graphene nanosheet with $CdLa_2S_4$ nanocrystals. The as-prepared samples show a higher efficiency for the photocatalytic degradation of typical MB dye compared with P25 and $CdLa_2S_4$ bulk nanoparticles. The enhancement of visible-light-responsive photocatalytic properties by decolorization of Rh.B dye may be attributed to the following causes. Firstly, graphene nanosheet is capable of accepting, transporting and storing electrons, and thus retarding or hindering the recombination of the electrons with the holes remaining on the excited $CdLa_2S_4$ nanoparticles. Secondly, graphene nanosheet can increase the adsorption of pollutants. The final cause is that their extended light absorption range. This work not only offers a simple way to synthesize graphene-based composites via a one-step process at low temperature but also a path to obtain efficient functional materials for environmental purification and other applications.

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.

1,3,5-Trichloro-2,4,6-Triazinetrion: A Versatile Heterocycle for the One-Pot Synthesis of 14-Aryl- or Alkyl -14H-Dibenzo[a,j]xanthene, 1,8-Dioxooctahydroxanthene and 12-Aryl-8,9,10,12-Tetrahydrobenzo[a]xanthene-11-one Derivatives under Solvent-Free Conditions

  • Maleki, Behrooz;Gholizadeh, Mostafa;Sepehr, Zeinalabedin
    • Bulletin of the Korean Chemical Society
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    • v.32 no.5
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    • pp.1697-1702
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    • 2011
  • A facile, green, efficient and environment-friendly protocol for the synthesis of 14-aryl- or alkyl-14Hdibenzo[a,j]xanthene, 1,8-dioxooctahydroxanthene and 12-aryl-8,9,10,12-tetrahydrobenzo[a]xanthene-11-one have been developed by one-pot condensation of various aldehydes with (i) ${\beta}$-naphthol (ii) cyclic 1,3-dicarbonyl compounds and (iii) ${\beta}$-naphthol and cyclic 1,3-dicarbonyl compounds, in the presence of 1,3,5-trichloro-2,4,6-triazinetrion (trichloroisocyanuric acid, TCCA) as catalyst under solvent-free conditions. The present approach offers the advantages of clean reaction, simple methodology, short reaction time, easy purification, and economic availability of the catalyst.