• 제목/요약/키워드: purification method

검색결과 848건 처리시간 0.03초

머캅탄류 검출을 위한 Thiobacillus thiooxidans가 생산하는 메칠머캅탄 산화효소의 분리 및 정제 (Isolation and Purification of Methyl Mercaptan Oxidase from Thiobacillus thiooxidans for Detection of Mercaptasn)

  • 김상준;신현재;이대실;양지원
    • KSBB Journal
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    • 제15권2호
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    • pp.145-149
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    • 2000
  • Thiobacillus thiooxidans KCTC2505에서 메칠머캅탄(MM) 가스를 기질로 메칠머캅탄 산화효소(MMO)를 유도할 수 있었으며 DEAE-Sephacel과 Superose 12 컬럼 크로마토그래피를 이용하여 정제하였다. 이때 얻어진 각 효소의 비활성은 각각 19.7과 80.1 units/mg-protein이었으며 효소의 최적 온도는 $43^{\circ}C$였다. 상기 정제된 효소액은 SDS-PAGE상에서 68.1 kDa의 분자량을 가진다. 이 효소는 암모늄염의 존재하에서는 활성이 증가하였으나 KCl 존재하에서는 감소하였으며 NaCl에 대해서는 거의 영향을 받지 않는 특성을 가지고 있다. 에탄올, 메탄올, 글리세린의 첨가에 대해 효소활성은 부분적으로 불활성화되었으며 아세톤의 경우에는 완전히 저해되었다. Purpald 발색법을 이용한 흡광도의 변화는 구강내 존재하는 MM 가스로부터 생성될 수 있는 수 십 nmol의 범위의 포름알데하이드에 대해 눈으로 인지가능한 발색시스템에 사용할 수 있음을 확인하였다.

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Identification of Histone Deacetylase 1 Protein Complexes in Liver Cancer Cells

  • Farooq, Muhammad;Hozzein, Wael N.;Elsayed, Elsayed A.;Taha, Nael A.;Wadaan, Mohammad A.M.
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권2호
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    • pp.915-921
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    • 2013
  • Background: Hepatocellular carcinoma is one of the leading causes of mortalities worldwide. The search for new therapeutic targets is of utmost importance for improved treatment. Altered expression of HDAC1 in hepatocellular carcinoma (HCC) and its requirement for liver formation in zebrafish, suggest that it may regulate key events in liver carcinogenesis and organogenesis. However, molecular mechanisms of HDAC1 action in liver carcinogenesis are largely unknown. The present study was conducted to identify HDAC1 interacting proteins in HepG2 cells using modified SH-double-affinity purification coupled with liquid mass spectrophotemetery. Materials and Methods: HepG2 cells were transfected with a construct containing HDAC1 with a C-terminal strepIII-HA tag as bait. Bait proteins were confirmed to be expressed in HepG2 cells by western blotting and purified by double affinity columns and protein complexes for analysis on a Thermo LTQ Orbitrap XL using a C18 nano flow ESI liquid chromatography system. Results: There were 27 proteins which showed novel interactions with HDAC1 identified only in this study, while 14 were among the established interactors. Various subunits of T complex proteins (TCP1) and prefoldin proteins (PFDN) were identified as interacting partners that showed high affinity with HDAC1 in HepG2 cells. Conclusions: The double affinity purification method adopted in this study was very successful in terms of specificity and reproducibility. The novel HDAC1 complex identified in this study could be better therapeutic target for treatment of hepatocellular carcinoma.

Improved Purification Process for Cholera Toxin and its Application to the Quantification of Residual Toxin in Cholera Vaccines

  • Jang, Hyun;Kim, Hyo-Seung;Kim, Jeong-Ah;Seo, Jin-Ho;Carbis, Rodney
    • Journal of Microbiology and Biotechnology
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    • 제19권1호
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    • pp.108-112
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    • 2009
  • A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

시설원예 배출 배액의 수질환경 평가 및 정화 부하량 산정 (The Water Quality and Purification Load Assessment of Drain Water of Facility Horticulture Areas)

  • 손진관;최덕규;공민재;윤성욱;박민정;강동현
    • 한국환경과학회지
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    • 제28권12호
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    • pp.1199-1208
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    • 2019
  • Korea's protected horticulture is rapidly increasing in scale due to various advantages such as year-round harvesting, labor savings through automation and shortened culture period, and greater income generation. This study was conducted to investigate the impact of protected horticulture on water quality. The results of this study are expected to provide basic data contributing to improvements towards sustainable agriculture and eco-friendly design of protected horticulture complex. The average T-N and T-P loads from vinyl greenhouses were 286.55± 143.98 mg/L and 59.14±13.77 mg/L, respectively and those from glass greenhouses 380.68 ± 150.41 mg/L and 61.85±20.72 mg/L. The annual discharge of wastewater derived from the monthly discharge from the horticulture greenhouses were estimated at 2597 ton/ha, with the annual phosphorus load amounting to 155.3 kg/ha. The average T-N and T-P loads in the tested greenhouse effluents were in excess of 8.3- and 13.5-fold the standards for the Korean wastewater plant effluent. The waste nutrient solution discharged from a protected horticulture complex can cause water contamination. Therefore, there is a need to conduct follow-up research using a water purification system or a trench method to develop a eco-friendly protected horticulture complex for sustainable agriculture.

키위열매 Protease 의 추출 정제 및 그 특성에 대하여 (Purification and Characterization of Kiwifruit Protease)

  • 김복자
    • 한국식품과학회지
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    • 제21권4호
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    • pp.569-574
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    • 1989
  • Kiwifruit에서 pretense를 추출 정제하여 그의 특성을 검토하였다. 조효소는 유안분획, sephadex G-100 filtration 및 DEAE-sephadex A-50 column chromatography를 거쳐 정제되었으며 정제효소의 비활성은 30.10으로 10.95배 증가하였고 활성수율은 7.48%에 달하였다. 정제효소는 casein및 hemoglobin을 잘 분해하였고 작용 최적 pH는 7.0이었으며 pH$7.0{\sim}8.0$에서 안정하였고 작용 최적온도는 $45^{\circ}C$이고 $50^{\circ}C$이하에서 안정하였다. 0.5 mM $HgCl_2$$MnSO_4$에 의해 강한 저해를 받았으며 2 mM cysteine과 0.5 mM EDTA에 의해 활성이 촉진되었으며 Km값은 50.5 mg/ml 이었고 분자량은 SDS 전기영동법에 의하여 측정하였을 때 23,500이었다.

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은 이온 크로마토그래피에 의한 오징어유로부터 eicosapentaenoic acid(EPA) 및 docosahexaenoic acid(DHA)의 분리농축 (Purification of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) Esters from Squid Oil by Silver Ion Chromatography)

  • 경영수;우영련;윤정로
    • 한국식품과학회지
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    • 제36권2호
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    • pp.361-364
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    • 2004
  • 은 이온이 불포화 지방산과의 착화합물을 만든다는 이론에 근거하여 은 이온 교환수지(SER)를 제조하였다. 제조한 SER을 비롯하여 silica gel, 질산은 함침 silica gel, 은 이온 제올라이트를 column 충전물로 사용하여 EPA와 DHA를 분리 농축하고 그 결과를 비교 분석하였다. SER과 silica gel의 9 : 1(w/w)혼합물을 충전물로 사용하였을 때 결과가 가장 좋았으며 이 경우 EPA와 DHA는 각각 27.9%와 49.1%로 농축되었고 수율은 각각 86.0%, 87.3%로 나타났다. SER만을 사용한 경우 EPA와 DHA는 각각 23.5%와 42.1%로 농축되었으며 이는 SER과 silica gel의 혼합 충전물 사용 시 보다 다소 낮은 결과이었다. 질산은 함침 silica gel의 경우 다른 충전물과 비교하여 농축율과 수율이 그다지 좋지 않았으나 EPA와 DHA의 분리 측면에서는 가장 우수한 결과를 나타내었다. SER은 재사용이 가능하고, 사용한 은 이온수지 자체도 쉽게 재생할 수 있을 뿐 아니라 사용한 은 이온도 AgCl 침전이나 $AgNO_3$로 회수가 용이하다는 점에서 다른 농축과정과 비교하여 훨씬 경제적이라 할 수 있다.

Investigation on purification of α-Fe2O3 from zinc smelting iron slag by superconducting HGMS technology

  • Zhang, Peng;Li, Su-qin;Guo, Zi-jie;Zhang, Chang-quan;Yang, Chang-qiao;Han, Shuai-shuai
    • 한국초전도ㆍ저온공학회논문지
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    • 제20권2호
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    • pp.16-19
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    • 2018
  • Comprehensive utilization of zinc smelting iron slag not only solves environmental problems but also creates huge economic benefits. This study was conducted on the enrichment and recovery of ${\alpha}-Fe_2O_3$ from zinc smelting iron slag by superconducting HGMS technology. Several variables such as slurry flow velocity, slag concentration, magnetic field intensity and the amount of dispersing agent were tested in magnetic separation. In the experiments, obtained optimal magnetic separation parameters were 1.60 T of magnetic flux intensity, 600 mL/min of slurry flow velocity of, 15 g/L of slag concentration of, 0.10 g/L of dispersing agent. Under this condition, the content of ${\alpha}-Fe_2O_3$ was increased from 86.22% to 94.39% that can approach the Chinese national standard requirements (A level) of iron oxide red. It was concluded that using superconducting HGMS technology was an effective method for the purification of ${\alpha}-Fe_2O_3$ from zinc smelting iron slag.

다공성 스테인리스 강 지지체의 표면개질에 따른 팔라듐-은 합금 수소 분리막의 수소 투과 선택도의 변화 (Effect of Surface Modification of the Porous Stainless Steel Support on Hydrogen Perm-selectivity of the Pd-Ag Alloy Hydrogen Separation Membranes)

  • 김낙천;김세홍;이진범;김현희;양지혜;김동원
    • 한국표면공학회지
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    • 제49권3호
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    • pp.286-300
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    • 2016
  • Pd-Ag alloy membranes have attracted a great deal of attention for their use in hydrogen purification and separation due to their high theoretical permeability, infinite selectivity and chemical compatibility with hydro-carbon containing gas streams. For commercial application, Pd-based membranes for hydrogen purification and separation need not only a high perm-selectivity but also a stable long-term durability. However, it has been difficult to fabricate thin, dense Pd-Ag alloy membranes on a porous stainless steel metal support with surface pores free and a stable diffusion barrier for preventing metallic diffusion from the porous stainless steel support. In this study, thin Pd-Ag alloy membranes were prepared by advanced Pd/Ag/Pd/Ag/Pd multi-layer sputter deposition on the modified porous stainless steel support using rough polishing/$ZrO_2$ powder filling and micro-polishing surface treatment, and following Ag up-filling heat treatment. Because the modified Pd-Ag alloy membranes using rough polishing/$ZrO_2$ powder filling method demonstrate high hydrogen permeability as well as diffusion barrier efficiency, it leads to the performance improvement in hydrogen perm-selectivity. Our membranes, therefore, are expected to be applicable to industrial fields for hydrogen purification and separation owing to enhanced functionality, durability and metal support/Pd alloy film integration.

Improved Purification of Thermophilic FoF1-ATP Synthase c-Subunit Rings and Solid-State NMR Characterization of Them in Different Lipid Membranes

  • Bak, Suyeon;Kang, Su-Jin;Suzuki, Toshiharu;Yoshida, Masasuke;Fujiwara, Toshimichi;Akutsu, Hideo
    • 한국자기공명학회논문지
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    • 제17권2호
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    • pp.67-75
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    • 2013
  • ATP synthase produces ATP, a major energy source for metabolic processes in organisms, from ADP and inorganic phosphate in cellular membranes. ATP synthase is known as a rotary motor, in which the c-subunit ring functions as a rotor. In this work, we have tried to develop a more general preparation procedure of thermophilic $F_oc$-ring ($TF_oc$-ring) for NMR measurements. The expression of $TF_oF_1$ is easily affected by various experimental conditions such as temperature, shape and size of a flask, a volume of medium, and shaking rate of an incubator. Accordingly, we have tried to optimize the expression conditions of $TF_oF_1$. $TF_oc$-rings were purified from $TF_oF_1$ according to a reported method. We modified purification procedures to improve purity and yield of $TF_oc$. On top of them, we found a new combination of detergents for the purification at anion-exchange column chromatography. To examine the effect of lipid environments on the structure, the $TF_oc$-rings were reconstituted into two kinds of lipid bilayers, namely, saturated and unsaturated lipid ones. Then, we have compared characteristics of the $TF_oc$-ring structures in these membranes with solid-state NMR.

메탄올 용매추출을 이용한 콜타르 흡수유 중의 인돌 결정화 (Indole Crystallization in Coal Tar Absorption Oil using Methanol Solvent Extraction)

  • 류희용;이상헌;신성순
    • 공업화학
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    • 제33권2호
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    • pp.166-172
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    • 2022
  • 콜타르 흡수유에 함유되어 있는 성분 중 고부가가치의 인돌을 효율적으로 분리, 회수 하기 위하여 추출-증류-결정화의 과정을 거치는 방법으로 천연 인돌 확보 기술을 연구하였다. 본 연구에 사용된 콜타르 흡수유의 주요 성분은 1.2% 나프탈렌, 0.1% 퀴놀린, 0.4% 이소퀴놀린, 6.4% 인돌, 21.0% 1-메틸나프탈렌, 48.8% 2-메틸나프탈렌 그리고 11.7% 비페닐을 포함하고 있다. 인돌의 분리, 정제를 위해 먼저 메탄올을 용매로 사용하여 콜타르 흡수유 중 인돌 성분을 추출상으로 분리한 후, 증류법으로 메탄올을 회수하였다. 계속해서 메탄올이 제거된 추출용액에 노말헥산을 혼합한 후 결정화하여 순도 99.3%의 인돌을 회수하였다. 또한 본 연구 결과를 바탕으로 개략적인 콜타르 흡수유 중 인돌 회수의 공정을 제안하였다.