• Title/Summary/Keyword: High-purity purification

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Improved Genomic DNA Isolation from Soil (토양으로부터 genomic DNA의 효과적인 분리)

  • Kang Ju-Hyung;Kim Bo-Hye;Lee Sun-Yi;Kim Yeong-Jin;Lee Ju-Won;Park Young Min;Ahn Soon-Cheol
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.851-856
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    • 2005
  • Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

Quality Improvement of Crude Glycerol from Biodiesel Production Using Activated Carbon Derived from Krabok (Irvingia malayana) Seed Shells

  • Wuttichai Roschat;Sarunya Donrussamee;Phatcharanan Smanmit;Samlit Jikjak;Tappagorn Leelatam;Sunti Phewphong;Krittiyanee Namwongsa;Preecha Moonsin;Vinich Promarak
    • Korean Journal of Materials Research
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    • v.34 no.1
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    • pp.1-11
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    • 2024
  • This research investigated the preparation of activated carbon derived from Krabok (Irvingia malayana) seed shells to improve the quality of crude glycerol obtained during biodiesel production. The activated carbon was prepared using a dry chemical activation method with NaOH, utilizing an innovative biomass incinerator. The results revealed that the resulting KC/AC-two-step exhibited favorable physicochemical adsorption properties, with a high surface area of 758.72 m2/g and an iodine number of 611.10 mg/g. These values meet the criteria of the industrial product standard for activated carbon No. TIS 900-2004, as specified by the Ministry of Industry in Thailand. Additionally, the adsorption efficiency for methylene blue reached an impressive 99.35 %. This developed activated carbon was then used to improve the quality of crude glycerol obtained from biodiesel production. The experimental results showed that the KC/AC-two-step increased the purity of crude glycerol to 73.61 %. In comparison, commercially available activated carbon (C/AC) resulted in a higher crude glycerol purity of 81.19 %, as analyzed by the GC technique. Additionally, the metal content (Zn, Cu, Fe, Pb, Cd, and Na) in purified glycerol using KC/AC-two-step was below the standards for heavy metals permitted in food and cosmeceuticals by the Food and Drug Administration of Thailand and the European Committee for Food Contact Materials and Articles. As a result, it can be inferred that Krabok seed shells have favorable properties for producing activated carbon suitable as an adsorbent to enhance crude glycerol purity. Furthermore, the improved crude glycerol from this research has potential for various industrial applications.

Corrosion control technique for pipeline system through injecting water stabilizer (수질안정화 약품 주입에 따른 상수도관 내부 부식제어 특성 연구)

  • Hwang, Byung-Gi;Woo, Dal-Sik
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.1
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    • pp.545-551
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    • 2011
  • Recently, demands for generating high quality tap waters are increasing with high concern of water pollution and corrosion of water pipelines. For the reasons, developing water quality stabilization technique in water purification system is sought rather than replacing to a new pipelines. In this study, high-purity liquid lime($Ca(OH)_2$) was introduced for a water quality stabilization technique in water purification process and simulated water distribution system of pilot-scale size was applied to evaluate anti-corrosion control effect. The effect of anti-corrosion control was calculated in terms of LSI(Langelier Saturation Index) In conclusion, the result of pilot plant showed improvement of corrosiveness by liquid lime($Ca(OH)_2$) with reduction of released iron(Fe). Application of anti-corrosion control technique to the mild steel coupon and the copper coupon were effective by indicating 35.4, 44.5% of improvements. Besides, sample pipes which were treated with liquid lime had formated more thicker layer of corrosion product inside of pipes. As a result, the process of injecting water stabilizer can greatly contribute to the high quality of tap water.

Preparation of Asymmetric PES Hollow Fiber Gas Separation Membranes and Their $CO_2/CH_4$ Separation Properties (비대칭구조의 폴리이서설폰 기체분리용 중공사막의 제조 및 이를 이용한 $CO_2/CH_4$ 분리특성)

  • Park, Sung-Ryul;Ahn, Hyo-Seong;Kim, Jeong-Hoon
    • Membrane Journal
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    • v.21 no.4
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    • pp.367-376
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    • 2011
  • Huge amount of $CH_4$ mixtures has been emitted from landfills and organic wastes via anaerobic digestion. The recovery of high purity $CH_4$ from these gases has two merits: reduction of green house gases and production of renewable fuels. Membrane technology based on polymeric materials can be used in this application. In this study, asymmetric gas separation hollow fiber membranes were fabricated to develop the membrane-based bio-gas purification process. Polyethersulfone (PES) was chosen as a polymer materials because of high $CO_2$ permeability of 3.4 barrer and $CO_2/CH_4$ selectivity of 50[1]. Acetone was used as a non-solvent additive because of its unique swelling power for PES and highly volatile character. The prepared PES hollow fiber showed excellent separation properties: 36 GPU of $CO_2$ permeance and 46 of $CO_2/CH_4$ selectivity at optimized preparation conditions: 9wt% acetone content, 10cm air-gap and 4wt% PDMS coating processes. With the PES hollow fiber membranes developed, mixed $CO_2/CH_4$ test was done by changing various operating conditions such as pressures and feed compositions to meet the highest recovery of CH4 with 95% purity. High $CH_4$ recovery of 58 wt% was observed at 10 atm feed pressure for the 50 vol% of $CO_2$ in $CO_2/CH_4$ mixture.

Preparation of High-grade Silica Sand for Metallurgical-grade Si Using a Physical Beneficiation (금속급 실리콘용 고순도 규사 제조를 위한 물리적 정제 특성)

  • Yang, Young-Cheol;Jeong, Soo-Bok;Chae, Young-Bae;Kim, Seong
    • Journal of the Mineralogical Society of Korea
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    • v.22 no.3
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    • pp.191-197
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    • 2009
  • It is very important to raise the purity of silica for manufacturing metallurgical-grade silicon because the purification of silicon in the smelting process is very difficult. In present study, the silica sand which is obtained from Vietnam was mineralogically analyzed. Based on the results, a novel process to separate impurities from the silica sand was developed, which consisted of classification, specific gravity and magnetic separation steps. Using the developed process, high-grade silica sand concentrate containing over 99.8 wt% $SiO_2$ was prepared, being suitable for manufacturing the metallurgical-grade silicon.

Microstructure Analysis of Fe Thin Films Prepared by Ion Beam Deposition (이온빔 증착법에 의해 제조된 철박막의 미세조직 분석)

  • Kim, Ka Hee;Yang, Jun-Mo;Ahn, Chi Won;Seo, Hyun Sang;Kang, Il-Suk;Hwang, Wook-Jung
    • Korean Journal of Metals and Materials
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    • v.46 no.7
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    • pp.458-463
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    • 2008
  • High purity Fe thin films were prepared by the ion beam deposition method with $^{56}Fe^{+}$ions on the Si substrate at the room temperature. The Fe thin films were deposited at the ion energy of 50 eV and 100 eV. Microstructural properties were investigated on the atomic scale using high-resolution transmission electron microscopy (HRTEM). It was found that the Fe thin film obtained with the energy of 50 eV having an excellent corrosion resistance consists of the amorphous layer of ~15 nm in thickness and the bcc crystalline layer of about 30 nm in grain size, while the thin film obtained with the energy of 100 eV having a poor corrosion resistance consists of little amorphous layer and the defective crystalline layer. Furthermore the crystal structures and arrangements of the oxide layers formed on the Fe thin films were analyzed by processing of the HRTEM images. It was concluded that the corrosion behavior of Fe thin films relates to the surface morphology and the crystalline structure as well as the degree of purification.

Comparative Study on Characterization of Recombinant B Subunit of E. coli Heat-Labile Enterotoxin (rLTB) Prepared from E. coli and P. pastoris

  • Ma, Xingyuan;Yao, Bi;Zheng, Wenyun;Li, Linfeng
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.550-557
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    • 2010
  • Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

Optimization and High-level Expression of a Functional GST-tagged rHLT-B in Escherichia coli and GM1 Binding Ability of Purified rHLT-B

  • Ma Xingyuan;Zheng Wenyun;Wang Tianwen;Wei Dongzhi;Ma Yushu
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.293-300
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    • 2006
  • The Escherichia coli heat-labile enterotoxin B subunit (HLT-B) is one of the most powerful mucosal immunogens and known mucosal adjuvants. However, the induction of high levels of HLT-B expression in E. coli has proven a difficult proposition. Therefore, in this study, the HLT-B gene was cloned from pathogenic E. coli and expressed as a fusion protein with GST (glutathion S-transferase) in E. coli BL2l (DE3), in an attempt to harvest a large quantity of soluble HLT-B. The culture conditions, including the culture media used, temperature, pH and the presence of lactose as an inducer, were all optimized in order to obtain an increase in the expression of soluble GST-rHLT-B. The biological activity of the purified rHLT-B was assayed in a series of GMI-ELISA experiments. The findings of these trials indicated that the yield of soluble recombinant GST-rHLT-B could be increased by up to 3-fold, as compared with that seen prior to the optimization, and that lactose was a more efficient alternative inducer than IPTG. The production of rHLT-B, at 92 % purity, reached an optimal level of 96 mg/l in a 3.7 L fermentor. The specific GM1 binding ability of the purified rHLT-B was determined to be almost identical to that of standard CTB.

Expression and Purification of Biologically Active Human Bone Morphogenetic Protein-4 in Recombinant Chinese Hamster Ovary Cells

  • Cha, Minyub;Han, Nara;Pi, Jia;Jeong, Yongsu;Baek, Kwanghee;Yoon, Jaeseung
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1281-1287
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    • 2017
  • Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of $EC_{50}$ of 2.93 ng/ml.

High Yield Production of Cyclofructan by Deletion Mutant Enzyme of Cycloinulooligosaccharide Fructanotransferase (Cycloinulooligosaccharide fructanotransferase의 결손변이효소에 의한 cyclofructan의 고효율 생산)

  • Park Jung-Ha;Kwon Hyun-Ju;Kim Byung-Woo
    • Journal of Life Science
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    • v.16 no.1
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    • pp.1-5
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    • 2006
  • This study investigated the optimal conditions of high yield production of cyclofructan (CF) using recombinant deletion mutant enzyme CFT108 which is constructed by N-terminal deletion from cycloinulooligosaccharide fructanotransferase (CFTase) gene of Penibacillus polymyxa. The production yield was dependent on reaction time, substrate concentration and enzyme concentration. The optimum reaction time for industrial purpose was achieved at 3 hr reaction. The optimal concentrations of substrate and enzyme were found to be $2\%$ inulin and 40 unit/ g inulin, respectively. At optimum condition, 9.5 g/l of maximum yield and $47.5\%$ of conversion efficacy were achieved. For purification of CF produced, the reaction mixture was treated with 1 unit/ml exoinulinase and then added $3\%$ CaO three times with blowing $CO_2$ gas, resulted in $95\%$ purity.