• 제목/요약/키워드: Purification method

검색결과 844건 처리시간 0.033초

Intein을 이용한 대장균에서의 Trichoderma reesei 유래의 Cellobiohydrolase I 섬유소 결합 도메인의 발현 (Intein-mediated expression of Trichoderma reesei Cellobiohydrolase I Cellulose Binding Domain in E. coli)

  • 최신건
    • 산업기술연구
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    • 제36권
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    • pp.33-37
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    • 2016
  • Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli. Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.

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Preliminary Experiment of Gravel Contact Oxidation Process in the Tropics

  • Abdullah Keizrul bin;Omachi Toshikatsu
    • 한국수자원학회:학술대회논문집
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    • 한국수자원학회 2006년도 학술발표회 논문집
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    • pp.12-17
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    • 2006
  • Natural rivers have water purification functions called Gravel Contact Oxidation Process, which decontaminate river water by biological absorption, oxidation and degradation on riverbed gravels. This function has been developed and applied to many small/medium-sized urban rivers in Japan as one of the direct river water purification methods. However the method hasn't been verified in the tropics, which have different climate conditions and river characteristics. A preliminary experiment carried out at a polluted urban tributary in the outskirts of Kuala Lumpur, Malaysia where an increasing attention has been paid to river environment, showed a good applicability to the tropical conditions as a technically practical water purification measure with some maintenance cares for sludge management.

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Concentration/Purification Technologies: Multi-Functionalities of Nanostructures in Biosensing Fields

  • Son, Sang Jun;Min, Junhong
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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    • pp.87-87
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    • 2013
  • Sample concentration and purification processes are essential in the bio-analytical and pharmaceutical fields because most bio samples or media are extremely sophisticated. To concentrate and purify specific substances, passive membrane type filters have been utilized, which is driven by size or charge differences between target and others. The traditional and representative method to identify nucleic acid sequences in the complex biosample is gel electrophoresis, which has been worked by size and net charge of molecules. The adsorption phenomena have been also utilized to concentrate and purify biomolecules. This adsorption of biomolecule can be controlled under specific salts and surfaces as well as surface area. To utilize the differences of physical properties of molecules or bio-targets such as virus, bacteria, and cells, the nanotechnologies can be introduced in target concentration, purification, and isolation processes. In here, I'd like to briefly survey typical examples of nanobiotechnologies which are introduced in sample treatment. Also I specifically demonstrate two different simple techniques to concentrate and detect bacteria from the samples using multifunctional silica nanotube (SNT).

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Purification and Characterization of Bacteriocin J105 Produced by Lactococcus latis subsp. lactis J105 Isolated from Kimchi

  • Kwak, Gyu-Suk;Kim, Sung-Koo;Jun, Hong-Ki
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.275-280
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    • 2001
  • Bacteriocin J105 is a proteinaceous inhibitory substance produced by Latococcus latis subsp. lactis j105 isolated from Kimchi. Bacteriocin J105 was purified to homogeneity by the pH-dependent adsorption-desorption method and reverse-phase HPLC from the culture broth of Lactococcus lactis subsp. lactis J105. Purification of bacteriocin J105 resulted in a 1.47-fold increase in the specific activity and the recovery was 1.5%. Its molecular mass measured by the electrophoretic pattern in the sodium, dodecyl sulfate polyacrylamide gel was about 3.4 kDa. It was stable at $121^{\circ}C$ for 15 min at pH between 2 and 4. However, at pH above 5, bacteriocin was rapidly inactivated. Twenty-one residues from the N-terminal portion of bacteriocin J105 were sequenced using sequence analysis of lantibiotics. Bacteriocin J105 showed significant homology with known nisin A from lactic acid bacteria.

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미나리꽝을 이용한 영양물질제거에 관한 연구 (A Studies on Removal of Nutrient Material by Using Dropwort Field)

  • 이영신;김창회
    • 한국환경보건학회지
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    • 제29권3호
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    • pp.16-20
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    • 2003
  • Nitrogen(N) and phosphorus(P) in surface streams mainly lead to euthrophication. It aggravates water quality and consequently increases the purification costs. As a resolution of water contamination caused by household drainage through irrigation route by 70% of the 1,300 community residents in Eum-Am Myun, Seo-San city, was implemented biological self-purification method by growing Oenanthe Javanica along the polluted water tunnel. The contaminated water was efficiently purified after passing the dropwort field; DO conc. of effluent water was increased 8.3∼61.9% after through the drop wort field. HRT of experiment system was changed 0.05∼1.50/day. 50% of BOD was eliminated at the range above 12 mg/l of Influent BOD conc. Also, 50% of COD was eliminated at the range above 30 mg/l of Influent COD conc. Finnally, the influent T-N loading at range below 1.5 g/m$^3$/d reduced 50% of Influent T-N conc., and so did influent T-P loading at the range below 0.03 g/m$^3$/dwas reduced 50% of Influent T-P conc.

A STUDY OF SIMULATION AND CONTROL OF PAC COSING PROCESS IN WATER PURIFICATION SYSTEM

  • Nahm, Euisuck;Lee, Subum;Woo, Kwangbang;Han, Taehan
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 1995년도 Proceedings of the Korea Automation Control Conference, 10th (KACC); Seoul, Korea; 23-25 Oct. 1995
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    • pp.75-78
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    • 1995
  • In this paper it is concerned to develop control method using jar-test results in order to predict the optimum dosage of coaglant, PAC(PoliAluminum Chloride). Considering the relations with the reactions with the reaction of coagulation and flocculation, the five independent variables ( e, g, turbidity of raw water, water turbidity in flocculators, temperature, pH, and alkalynity) are selected out of parameters and they are put into calculation to develop a neural network model for PAC dosing process in water purification system. This model is utilized to predict optimum dosage of PAC. That is, the optimum dosage of PAC is searched in neural network model for PAC dosing process to minimize the water turbidity in flocculators. This searching is implemented by means of expert heuristics. The efficacy of the proposed contorl schemem and feasibility of acquired neural network model for PAC dosing contorl in water purification system is evaluated by means of computer simulation.

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Characterization of Partially Purified Extracellular Protease of Local Bacteria BAC-4

  • Setiasih, Siswati
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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    • pp.161-161
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    • 1998
  • To achieve the aim of this investigation, the extracellular protease was isolated from bacteria BAC-4, a strain was cultivated in the medium for the production of penicillin acilase in a period of 32 hours. The enzyme was first purified by aceton precipitation method, followed by ion exchange chromatography on DEAE-sephacel column. The highest specific activity of the aceton fraction was found to be 2.19 unit per mg, with degree of purification of 13 times. Further purification of the enzyme on DEAE -sephacel had a specific activity of 58.6 unit per mg and degree of purification of 344 times compared to its crude extract. The optimum pH of the enzyme was 8.4, and the potimum temparature was 37$^{\circ}C$. The K$\_$M/ and $V_{max}$ calculated at experiment conditions were found to be 0.66%(W/V) and 3.61 unit per mL respectively.

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Application of Membrane Technology in Japan

  • Minami, Katsuyoshi
    • 한국막학회:학술대회논문집
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    • 한국막학회 1995년도 추계 총회 및 학술발표회
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    • pp.58-73
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    • 1995
  • Membrane filtration is a promising technology for efficient solid/liquid separation in water purification. In FY 1991, the Ministry of Health and Welfare, Japanese Government launched a comprehensive research project "MAC 21" for development of membrane technology and its application to public water supply. The project was conducted by the Water Purification Process Association (WPPA), under the supervision of the Institute of Public Health. By the research project from FY 1991 to FY 1993, we confirmed that microfiltration (MF)/ultrafiltration (UF) technology was applicable to water purification and MF/UF was a effective method for the removal of such contaminants as particulate matter and coliforms. The Guideline Committee organized under the Technical Committee prepared a the guidelines on application of membrane system to small-scale public water supplies, based on the results as written above. The guidelines has been published in Dec., 1994 by WPPA.4 by WPPA.

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A Novel Simple Method to Purify Recombinant Soluble Human Complement Receptor Type 1 (sCR 1) from CHO Cell Culture

  • Wang, Pi-Chao;Hisamune Kato;Takehiro Inoue;Masatoshi Matsumura;Noriyuki Ishii;Yoshinobu Murakami;Tsukasa Seya
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권2호
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    • pp.67-75
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    • 2002
  • The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.

Purification of Deoxycytidine Kinase from Various Human Leukemic Cells by End-product Analog Affinity Chromatography

  • Kim, Min-Young
    • BMB Reports
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    • 제28권4호
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    • pp.281-289
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    • 1995
  • Homogeneous human deoxycytidine kinase was purified in one step from a variety of spontaneous human leukemic cells (T-ALL, B-ALL, B-CLL, AML, CML), and from cultured T-lymphoblast cells (MOLT-4) using the newly developed affinity medium, $dCp_4$-Sepharose. Starting with an ammonium sulfate fraction, purification was achieved in one step with the kinase being eluted from a column by the end product inhibitor, dCTP. The purified deoxycytidine kinase from T-ALL cells phosphorylated deoxyadenosine and deoxyguanosine, as well as deoxycytidine. The enzyme purified from T-ALL and B-CLL cells yielded one major band with a molecular weight of 52 kDa determined by SDS-polyacrylamide gel electrophoresis. AML and CML cells yielded one 52 kDa band and an extra band of 30 kDa molecular weight. On the other hand, B-ALL and MOLT-4 cells showed a low molecular weight band of 30 kDa only. However, the electrophoretic mobilities of enzymatic activity in 12% non-denaturing gels were identical for the dCyd kinase from all different kinds of leukemic cell lines, except that the B-ALL, B-CLL, and MOLT-4 cell preparations had an extra minor peak, all at the same position. dAdo and dCyd phosphorylating activities comigrated indicating that these activities are all associated with the same protein. Two new methods, a disk implantation method and a nitrocellulose powder method were used with a small amount of enzyme protein to raise polyclonal antibodies against dCyd kinase purified from T-ALL cells.

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