• 제목/요약/키워드: High-purity purification

검색결과 159건 처리시간 0.025초

Novel Modification of Growth Medium Enables Efficient E. coli Expression and Simple Purification of an Endotoxin-Free Recombinant Murine Hsp70 Protein

  • Zachova, Katerinat;Krupka, Michal;Chamrad, Ivo;Belakova, Jana;Horynova, Milada;Weigl, Evzen;Sebela, Marek;Raska, Milan
    • Journal of Microbiology and Biotechnology
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    • 제19권7호
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    • pp.727-733
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    • 2009
  • Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

식물세포 배양으로부터 Peroxidase 대량 정제를 위한 전처리 공정 개발 (A New large-scale Pre-purification for Peroxidase from Plant Cell Cultures)

  • 표상현
    • KSBB Journal
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    • 제15권4호
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    • pp.342-345
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    • 2000
  • 식물세포배양으로부터 peroxidase를 대량 생산하기 위한 분리/정제 공정 으로서 세포를 파쇄하고 크로마토그래피 전처리로서 활성백토를 적용하였다. 활성백토는 미세한 세 포 조각 뿐만 아니라 여려가지 불순불을 선택적으로 흡착 하는 성질을 나타내었으며, 이를 통하여 효과적으로 정제 공정을 진행할 수 있었다 흡착을 통한 전처리 후 한외여 과장치를 이용하여 농축을 실시 하였으며, DEAE-Sepharose F FF를 이용한 크로마토그래피를 통해 정제가 이루어졌다. 활성을 나타내는 용액을 탈염, 농축 후 동결 건조하였다. 이 공정은 상업화에 필요한 대량 정제를 가능하게 하였으며 수율과 정제비용 측면에서 상당히 경제적인 공정임을 입증하였다, 활성백토를 이용한 흡착공정은 다른 효소 및 단백질의 경제적인 대량정제에 적용될 수 있으리라 기대된다.

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Mineralogical Analysis and Mechano-Chemical Purification of Natural Silica Ore for High Purity Silica Powder

  • Park, Jesik;Lee, Churl Kyoung;Lee, Hyun-Kwon
    • 한국재료학회지
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    • 제26권6호
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    • pp.306-310
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    • 2016
  • To produce 4N grade high-purity silica powder from natural ore, the mineralogical characteristics of natural silica ore were investigated and their effects on the purification process were revealed. The Chinese silica mineral ore used was composed of iron and aluminum as main impurities and calcium, magnesium, potassium, sodium, and titanium as trace impurities; these trace impurities generally exist as either single oxides or complex oxides. It was confirmed that liberation and acidic washing of the impurities were highly dependent on the particle size of the ground silica ore and on its mineralogical characteristics such as the distribution and phases of existing impurities. It is suggested that appropriate size reduction of silica ore should be realized for optimized purification according to the origin of the natural silica ore. A single step purification process, the mechano-chemical washing (MCW) process, was proposed and verified in comparison with the conventional multi step washing process.

Microbiological Purification of L-Arabitol from Xylitol Mother Liquor

  • Jiang, Mingguo;Wang, Ben;Yang, Lifang;Lin, Shuangjun;Cheng, Hairong
    • Journal of Microbiology and Biotechnology
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    • 제21권1호
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    • pp.43-49
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    • 2011
  • As a rare sugar alcohol, L-arabitol can be used in food and can prevent extra fat deposits in the intestinal tract. Commercially, L-arabitol is prepared from pure L-arabinose by hydrogenation, which needs a high temperature and high pressure, leading to a high production cost for Larabitol. Therefore, this study describes a novel L-arabitol production method based on biological purification from the xylitol mother liquor, a cheap and readily available raw material that contains a high concentration of Larabitol. First, a novel Bacillus megaterium strain was screened that can utilize xylitol, sorbitol, and mannitol, yet not L-arabitol. The isolated strain was inoculated into a medium containing the xylitol mother liquor under formulated culture conditions, where a high L-arabitol yield (95%) and high purity (80%) were obtained when the medium was supplemented with 50 g/l of xylitol mother liquor. Upon further purification of the fermentation broth by ion exchange and decolorization, L-arabitol was crystallized with a purity of 98.5%.

Preparation of High-Purity Urokinase Using Single-Step Hydrophobic Interaction Chromatography with p-Aminobenzamidine Ligand

  • Cao, Xue-Jun;Zhou, Jian-Hua;Huang, Zhen-Hui;Wu, Xing-Yan;Hur, Byung-Ki
    • Journal of Microbiology and Biotechnology
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    • 제12권2호
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    • pp.196-203
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    • 2002
  • A novel process for urokinase purification was studied using p-aminobenzamidine as the ligand and sepharose 4B as the matrix. The adsorption, washing, and elution conditions were optimized by an unusual method. An adsorption buffer containing 2.5 M NaCl and $1\%$ Tween 80 facilitated the adsorption of urokinase on the affinity media and prevented contaminants from binding to the p-aminobenzamidine affinity gel. It was found that $5\%$ Tween 80 removed most of the contaminants from the affinity column. A 0.2 M glycine elution buffer containing 0.5 M NaCl (pH 3.0) was found to have a strong elution ability with a high recovery and purity of urokinase. A crude urokinase material of231 IU/mg protein from human urine was purified to 124,300 IU/mg protein with a purification factor of 538 and yield of $86.7\%$. As a result, a high purity urokinase was obtained with only a single affinity chromatography step. The purification process was successfully scaled-up to a 2-1 chromatography column. The resulting urokinase eluate could be directly lyophilized, thereby complying with Chinese pharmacopoeia (1995 version) standards.

4탑 PSA 공정의 의한 SMR off-gas로부터 수소 정제 (Hydrogen Purification by the Four-Bed Pressure Swing Adsorption Process from Steam Methane Reforming Off-Gas)

  • 양세일;박주용;장성철;김성현;최대기
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2008년도 추계학술대회 논문집
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    • pp.383-386
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    • 2008
  • The four-bed PSA process using a layered bed of activated carbon and zeolite 5A was studied to produce a high purity hydrogen product from SMR off-gas. At a desired product purity (99.999%+), the recovery increased with decreasing the linear velocity. However, the difference of the increasing of the recovery became smaller with the decreasing of the linear velocity and then was similar from below the linear velocity 3.9 cm/s. When the adsorbents, the feed gas composition, and the operating conditions are given, the residence time is mainly a function for design of the PSA bed size. The minimum residence time exists to obtain the maximum recovery at desired product purity.

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콜타르 유분 중에 함유된 인돌의 고순도 정제 - 용액 결정화에 의한 인돌 유사 비점 혼합물의 분리 - (High-Purity Purification of Indole Contained in Coal Tar Fraction - Separation of Close Boiling Mixtures of Indole by Solute Crystallization -)

  • 김수진;강호철;정화진
    • 공업화학
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    • 제21권2호
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    • pp.238-241
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    • 2010
  • 콜타르 유분(유출온도범위: $240{\sim}265^{\circ}C$)으로부터 추출-증류의 조합에 의해 회수한 54.3 wt% 인돌 유출액(유출온도범위 $250{\sim}255^{\circ}C$)을 원료로 사용하여 용액 결정화 조작을 행해 인돌의 고순도 정제를 검토했다. 원료 유출액은 8성분(quinoline, iso-quinoline, indole, quinaldine, 1-methylnaphthalene, 2-methylnaphthalene, biphenyl, phenyl ether)으로 구성되어 있다. 결정화 조작의 용매로서는 노말헥산을, 냉매로서는 메탄올 수용액(50 : 50 vol%)을 각각 사용하였으며, 결정화 장치로서는 재킷이 부착된 pyrex제 유리 회분 교반기를 사용했다. 결정화 온도와 용매/원료의 초기 체적비의 증가는 인돌의 순도를 향상시켰으나, 역으로 수율을 저하시켰다. 석출된 결정의 세정조작 없이, 노말헥산을 용매로 사용한 용액 결정화 조작에 의해 고순도(99.3 wt%)의 인돌을 고수율(50%)로 회수할 수 있었다.

해조류로부터 푸코이단/알지네이트의 고순도 분리정제 (Methods for High Quality Purification of Alginate and Fucoidan from Marine Brown Algae)

  • 김종기;양재호
    • 한국해양바이오학회지
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    • 제2권4호
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    • pp.245-249
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    • 2007
  • High quality of purified alginate and fucoidan is required for the medical uses to prevent the unexpected side effects from the contaminated endotoxin present in the materials. We attempted to establish an efficient and fast mass scale production method for the highly purified poly-G alginate from the sea weeds. In addition, lab scale method was established to obtain the fraction of higher purity from the commercially available fucoidan source (about 85% purity).

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암모늄 디나이트라마이드염의 합성 및 액상연료화 연구 (Preparation of High Purity Ammonium Dinitramide and Its Liquid Mono-propellant)

  • 김우람;박미정;김소희;전종기;조영민
    • 공업화학
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    • 제30권5호
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    • pp.591-596
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    • 2019
  • 친환경 대체연료물질로서 ammonium dinitramide (ADN, $NH_4N(NO_2)_2$)는 상온에서 안전하고, 안정하지만 실제 적용을 위해서는 고순도가 요구된다. 소량의 불순물은 단일계 액상연료용 추력기 내 촉매의 분해반응을 억제하며, 비추력을 저하시키고, 노즐 막힘과 같은 부작용을 초래한다. 따라서 본 연구는 반복추출, 활성탄에 의한 흡착, 그리고 저온추출방법을 적용하여 합성한 ADN을 정제하였고, FT-IR, UV-Vis 및 IC 분석을 통하여 화학적인 순도를 평가하였으며, 최종 순도로서 IC 분석기준으로 99.82%를 획득하였다. 또한, ADN을 주 산화제로 활용하는 액상연료를 제조하였으며, 최소 $148^{\circ}C$에서 분해되는 단일계 추진제를 합성하였다. 그러나 상안정화를 위하여 우레아를 연료물질로 추가하였을 경우, 분해온도는 $188^{\circ}C$까지 상승하였다.

Electrical Properties of Transparent Conductive Films of Single-Walled Carbon Nanotubes with Their Purities

  • ;;;이내성
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2010년도 하계학술대회 논문집
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    • pp.56-56
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    • 2010
  • Single-walled carbon nanotubes (SWCNTs) have attracted much attention as a promising material for transparent conducting films (TCFs), due to their superior electrical conductivity, high mechanical strength, and complete flexibility as well as their one-dimensional morphological features of extremely high length-to-diameter ratios. This study investigated three kinds of SWCNTs with different purities: as-produced SWCNTs (AP-SWCNTs), thermally purified SWCNTs (TH-SWCNTs), thermally and acid purified SWCNTs (TA-SWCNTs). The purity of each SWCNT sample was assessed by considering absorption peaks in the semiconducting ($S_{22}$) and metallic ($M_{11}$) tubes with UV-Vis NIR spectroscopy and a metal content with thermogravimetric analysis (TGA). The purity increased as proceeding the purification stages from the AP-SWCNTs through the thermal purification to the acid purification. The samples containing different contents of SWCNTs were dispersed in water using sodium dodecyl benzensulfate (SDBS). Aqueous suspensions of different purities of SWCNTs were prepared to have similar absorbances in UV-Vis absorption measurements so that one can make the TCFs possess similar optical transmittances irrespective of the SWCNT purity. Transparent conductive SWCNT networks were formed by spraying an SWCNT suspension onto a poly(ethyleneterephthalate) (PET) substrate. As expected, the TCFs fabricated with AP-SWCNTs showed very high sheet resistances. Interestingly, the TH-SWCNTs gave lower sheet resistances to the TFCs than the TA-SWCNTs although the latter was of higher purity in the SWCNT content than the former. The TA-SWCNTs would be shortened in length and be more bundled by the acid purification, relative to the TH-SWCNTs. For both purified (TH, TA) samples, the subsequent nitric acid ($HNO_3$) treatment greatly lowered the sheet resistances of the TCFs, but almost eliminated the difference of sheet resistances between them. This seems to be because the electrical conductivity increased not only due to further removal of surfactants but also due to p-type doping upon the acid treatment. The doping effect was likely to overwhelm the effect of surfactant removal. Although the nitric acid treatment resulted in the similar. electrical properties to the two samples, the TCFs of TH-SWCNTs showed much lower sheet resistances than those of the TA-SWCNTs prior to the acid treatment.

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