• Title/Summary/Keyword: one-step purification

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Efficient Method for the Rapid Purification of Nosema ceranae Spores

  • Kim, Dong-Jun;Yun, Hwi-Geon;Kim, In-Hui;Gwak, Won-Seok;Woo, Soo-Dong
    • Mycobiology
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    • v.45 no.3
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    • pp.204-208
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    • 2017
  • Nosema ceranae is an obligate intracellular fungal parasite that causes mortality in honey bees and enhances the susceptibility of honey bees to other pathogens. Efficient purification of Nosema spores from the midgut of infected honey bees is very important because Nosema is non-culturable and only seasonably available. To achieve a higher yield of spores from honey bees, in this study, we considered that the initial release of spores from the midgut tissues was the most critical step. The use of 2 mm beads along with enzymatic treatment with collagenase and trypsin enhanced the homogenization of tissues and the yield of released spores by approximately 2.95 times compared with the use of common 3 mm beads alone. The optimal time for the enzyme treatment was determined to be 1 hr as measured by the yield and viability of the spores. A one-step filtration using a filter paper with an $8-11{\mu}m$ pore size was sufficient for removing cell debris. This method may be useful to purify not only N. ceranae spores but also other Nosema spp. spores.

Improved sulfur hexafluoride(SF6) gas analysis method for quality management of cryogenic refinement system (극저온 정제시스템의 품질관리를 위한 SF6 가스 분석방법 개선)

  • Lee, Jeong Eun;Cho, Min-ho;Lee, Won Seok
    • KEPCO Journal on Electric Power and Energy
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    • v.8 no.1
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    • pp.37-41
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    • 2022
  • Because sulfur hexafluoride(SF6) is classified as one of the six major greenhouse gases, SF6 handling in power plant such as recovery, purification, and reuse is considered to be important. KEPCO has focused to develop the advanced recovery and purification technology of SF6 reuse. SF6 analysis includes the on-site analyses and on-line analyzer; i.e., (1) on-site analysis has an error rate of ±0.5% and (2) on-line analysis has an error rate of ±0.1%, which is possible to adjust operating conditions and to make the work more conveniently by analyzing SF6 concentration before and after purification step. This paper presents an online analysis method in the SF6 purification and reuse system. In addition, the analysis results and quality guarantees for each section of the analysis system were presented.

Simple Purification of Escherichia coli-Derived Recombinant Human Interleukin-2 Expressed with N-terminus Fusion of Glucagon

  • Won Hye-Soon;Lee Jeewon;Kim In-Ho;Park Young-Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.13-16
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    • 2000
  • Simple procedures have been devised for purifying recombinant human interleukin-2 (hIL-2), which was expressed in Escberichia coli using sequences of glucagon molecules and enterokinase cleavage site as an N-terminus fusion partner. The insoluble aggregates of recombinant fusion protein produced in E. coli cytoplasm were easily dissolved by simple alkaline pH shift $(8\rightarrow12\rightarrow8)$. Following enterokinase cleavage, the recombinant hIL-2 was finally purified by one-step reversed-phase HPLC with high purity. The ease and high efficiency of this simple purification process seem to mainly result from the role of used glucagon fusion partner, which could be applied to the production of other therapeutically important proteins.

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One-step purification and biochemical characterization of a (s)-stereospecific esterase from Pseudomonas fluorescens KCTC 1767

  • Choe, Gi-Seop;Kim, Ji-Hui;Kim, Ji-Yeon;Kim, Geun-Jung;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.445-448
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    • 2002
  • The Pseudomonas fluorescens KCTC 1767, a selected and identified as potential candidate for stereo-specific resolution of rac-ketoprofen ethyl ester, was systematically investigated in order to induce the high level expression and detailed characterization of the expressing enzyme esterase. We cloned the esterase gene from chromosomal DNA of Pseudomonas fluorescens KCTC 1767 by PCR with two synthetic primers that desinged for simple purification. The recombinant esterase from Pseudomonas fluorescens KCTC 1767 exibited a high conversion rate and enantioselectivity to the (S)-ketoprofen ethyl ester as expected. The enzyme was easily purified to homogeniety by using a metal chelating affinity chromatography as a protein with poly histidine taq, and thus obtained 0.6 mg of protein from a 100 mL culture broth in a single step. The purified enzyme was steadily stable at the pH range from 7.0 to 10. The activity was also retained to be about 70% after the preincubation at $40^{\circ}C$ but over $50^{\circ}C$ lost the activity completely. The molecular mass of the esterase was estimated to be about 43 kDa on SDS-PAGE, and an identical result was also shown in gel filteration chromatography. The specific activity was calculated 27 mM/mg-protein/min by using the rac-ketoprofen ethly ester as a substrate.

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An Efficient Procedure for the Synthesis of Benzimidazoles Using H2O2/SiO2-FeCl3 System (H2O2/SiO2-FeCl3 계를 이용한 Benzimidazoles의 효율적인 합성)

  • Fazlinia, Abbas;Mosslemin, Mohammad Hossein;Sadoughi, Hesamaddin
    • Journal of the Korean Chemical Society
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    • v.54 no.5
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    • pp.579-581
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    • 2010
  • Solid silica supported ferric chloride ($SiO_2-FeCl_3$) catalyzed one-step synthesis of various benzimidazoles from o-phenylenediamine and aldehydes using $H_2O_2$ as the oxidant. The salient features of this method are simple and convenient procedure, easy purification and shorter reaction times.

Purification of a Mosquitocidal Toxic Protein from B. thuringiensis strain H9B by Immuno-Affinity Chromatography (Immuno-Affinity Chromatography에 의한 B. thuringiensis H9B 균주의 모기살충성 내독소 단백질의 정제)

  • 김광현;배수장;이광배
    • Journal of environmental and Sanitary engineering
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    • v.12 no.2
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    • pp.59-64
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    • 1997
  • For purification of a 70kDa toxic protein of mosquitocidal delta-endotoxin from B. thuringiensis strain H9B, immuno-affinity chromatography was performed. After separation of 70kDa toxic proteins from the delta-endotoxin of the strain H9B on SDS-PAGE, the 70kDa toxic protein was subcutaneously injected into rabbit for making a polyclonal antibody. A anti-70kDa toxic protein was purified by a column chromatography packed with protein A-sepharose 4B gels. The 70kDa toxic protein from delta-endotoxin of the strain H9B was also purified by an immuno-affinity chromatography packed with CNBr-activated sepharose 4B gels conjugated anti-70kDa toxic protein after elution with 1/10M citric acid-1/5M Na$_{2}$HPO$_{4}$ buffer(pH3.2) containing 0.5M NaCl. The 70kDa toxic protein was purified through only one step-separation system, was demonstrated by SDS-PAGE and immunoblot.

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Purification of Single-walled Carbon Nanotubes by HCl Treatment and Analysis of the Field Emission Property (염산에 의한 단중벽 탄소나노튜브 정제와 전자방출 특성 평가)

  • Lyu, SeungChul;Jung, Dami;Ahn, KiTae;Lee, Hansung;Lee, Naesung;Park, Yunsun;Sok, Junghyun
    • Korean Journal of Metals and Materials
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    • v.48 no.4
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    • pp.335-341
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    • 2010
  • High-quality single-walled carbon nanotubes (SWCNTs) were synthesized by catalytic decomposition of $C_2H_2$ using Fe-Mo/MgO catalyst at $800^{\circ}C$. The as-synthesized SWCNTs typically occurred in the form of a bundle with a diameter of 10~20 nm together with amorphous carbon and catalytic impurities, which were removed by a two-step purification process consisting of oxidation and an acid treatment. The oxidation step, using an $O_2$-Ar mixture at $380^{\circ}C$ for 5 hr in a vertical-type furnace and a $HNO_3$ treatment at $100^{\circ}C$ for one hour, was utilized to remove the amorphous carbon particles. Subsequently, metallic catalysts were removed in HCl at room temperature for 5 hr under magnetic stirring. The SWCNT suspension was prepared by dispersing the purified SWCNTs in an aqueous sodium dodecyl benzene sulfonate solution with horn-type sonication. This was then air-sprayed on glass to fabricate CNT field emitters. The samples had a turn-on field value of 4 V/${\mu}m$ and a current density of 0.67 mA/$cm^2$ at 9 V/${\mu}m$. Increasing the HCl treatment time improved the field emission properties.

Microbial production of coenzyme Q10

  • Suh, Jung-Woo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2006.11a
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    • pp.127-130
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    • 2006
  • Coenzyme Q10(CoQ10) is a biological quinine compound that is widely found in living organisms including yeast, plants, and animals. CoQ10 has two major physiological activities:(a)mitochondrial electron-transport activity and (b )antioxidant activity. Various clinical applications are also available: Parkinson's disease, Heart disease, diabetes. Because of its various application filed, the market size of CoQ10 is continuously expanding all over the world. A Japanese company, Nisshin Pharma Inc. is the first industrial producer of CoQ10(1974). CoQ10 can be produced by fermentation and chemical synthesis. In several companies, these two methods are used for the production of CoQ10:chemical synthesis - Yungjin, Daewoong, Nishin Parma; fermentation - Kaneka, Kyowa, Yungjin, etc. Researchs in microbial production of CoQ10 have several steps: screening of producing microorganisms, strain development, fermentation process, purification process, scale-up process, plant production. Several strategies are available for the strain development : Random mutation and screening, directed metabolic engineering. For the optimization of fermentation process, various conditions (nutrient, aeration, temperature, culture type, etc.) are considered. Purification is one of the most important step because the quality of final products entirely depends on its purity. The production cost will be reduced and the quality of the CoQ10 will be impoved by continuous researches in strain development, fermentation process, purification process.

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Expression and Purification of GFPuv/Cytochrome c-552 Fusion Protein in E. coli

  • Hong, Eul-Jae;Lee, Sang-On;Choe, Jeong-U;Hong, Eok-Gi
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.550-553
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    • 2003
  • The genes of GFPuv and Cytochrome c-552 were amplified by using PCR, and then, fused each other. Fusion gene of GFPuv and Cytochrome c-552 was inserted into the pTrcHis B vector and transferred to E. coli. A fusion protein of GFPuv and Cytochrome c-552 was expressed in JM109 and BL21. This fusion protein was composed of a His-tag for the rapid one-step purification using an immobilized metal affinity chromatography.

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Microbial production of coenzyme Q10

  • Suh, Jung-Woo
    • 한국약용작물학회:학술대회논문집
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    • 2006.11a
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    • pp.127-130
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    • 2006
  • Coenzyme Q10(CoQ10) is a biological quinine compound that is widely found in living organisms including yeast, plants, and animals. CoQ10 has two major physiological activities:(a)mitochondrial electron-transport activity and (b)antioxidant activity. Various clinical applications are also available : Parkinson's disease, Heart disease, diabetes. Because of its various application filed, the market size of CoQ 10 is continuously expanding all over the world. A Japanese company, Nisshin Pharma Inc. is the first industrial producer of CoQ10(1974). CoQ10 can be produced by fermentation and chemical synthesis. In several companies, these two methods are used for the production of CoQ10:chemical synthesis - Yungjin, Daewoong, Nishin Parma; fermentation - Kaneka, Kyowa, Yungjin, etc. Researchs in microbial production of CoQ10 have several steps: screening of producing microorganisms, strain development, fermentation process, purification process, scale-up process, plant production. Several strategies are available for the strain development : Random mutation and screening, directed metabolic engineering. For the optimization of fermentation process, various conditions (nutrient, aeration, temperature, culture type, etc.) are considered. Purification is one of the most important step because the quality of final products entirely depends on its purity. The production cost will be reduced and the quality of the CoQ10 will be impoved by continuous researches in strain development, fermentation process, purification process.

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