• Korean Journal of Microbiology
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• v.49 no.1
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• pp.95-98
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• 2013

Following construction of expression vectors in Escherichia coli, a new procedure involving two-step column purifications with a Fast Performance Liquid Chromatography System was developed for purification of the oxygenase component of alkylphenol hydroxylase of Pseudomonas alkylphenolia. From 50 g wet cake of recombinant E. coli BL21(DE3)(pJJPMO2) cells, 110 mg of pure protein in a heterodimeric form containing a stoichiometric amount of iron were obtained and it exhibited a specific activity of 147 nmole/min/mg.

• Proceedings of the Membrane Society of Korea Conference
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• 1996.10a
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• pp.77-79
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• 1996

근래들어 생물기술이 급속히 발전함에 따라 생물반응을 통해 다양한 생물제품들의 생산이 이루어지고 있다. 생물반응에 의한 생물제품들의 생산량은 극단적으로는 기질 1g당 10$^{-8}$g 정도로서 극히 낮아 이를 상업제품이 요구하는 순도(95 %이상)로 까지 정제하기 위해서는 여러 단계를 거치는 복잡하고 지루한 분리.정제 과정이 필요하며, 이 분리.정제의 후류공정(downstream process)비용이 생물제품 생산비의 상당부분을 점유케 되어 생물공정의 경제성을 낮게 한다. 따라서 생물공정을 이용한 생물제품 생산이 산업적으로 경제성을 갖기 위해서는 생물제품을 보다 효율적.경제적으로 분리.정제할 수 있는 후류공정의 확립이 요구된다. 본 연구에서는 산업적 규모로 생산되는 생물제품들을 효율적으로 분리.정제하는데 응용 가능한 방법의 하나로서 관심의 대상이 되고 있는 책체 크로마트그래피 기법을 연구의 대상으로 하였다.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.135-135
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• 1993

신호전달과정에 중요한 역할을 하고 있는 다기능 serinei/threonine 단백질인산화효소인 protein kinase C(PKC)의 연구를 위해 이 효소의 정제를 뇌에서 착수하였다 PKC의 활성측정을 myelin basic protein을 기질로 하여 20 mM Tris 완충용액 PH 7.5, 0.15 mM [${\gamma}$-$^{32}$P]ATP(3 $\times$ $10^{5}$ cpm), 0.1 mM $Ca^{2＋}$, 10$\mu\textrm{g}$ phosphatidylserine과 2$\mu\textrm{g}$ diolein을 넣어 반응시켰다. 반응은 TCA로 정지시킨 후 방사성 단백질을 Millipore filter paper로 걸러 섬광 계수기로 읽었다. Cytosol PKC의 정제과정은 첫 단계에서 DEAE-cellulose를 사용하였으며, phenyl sepharose CL-4B와 protamine agarose를 연속적으로 이용하여 800배의 정제에 성공했다. SDS-PACE 상에서 80 kD로 나타났으며 순도는 95 % 이상이였다. 이를 이용 PKC의 각종 기질 연구에 착수하기 시작했으며, 이중 MBP의 인산화연구를 통한 myelin의 안정성과 MBP와의 구조 관계가 일부 수행되고 있다 연차적으로 PKC와 이와 관련된 단백질의 특성을 살피기 위해 뇌의 PKC 기질 중 cold stress를 통해 환경에 민감한 것을 찾고 있으며, 현재 autoradiography를 이용해 80 kD, 54 kD, 49 kD와 35 kD의 단백질이 연구대상이 되고있다. 그 중 49 kD는 B-50(또는 GAP43, neuromodulin이라고도 함)일 가능성이 높아 이 단백질 조절과 PKC 활성화 사이의 관계 정립이 흥미로운 과제로 대두되고 있다.다.

• Membrane Journal
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• v.34 no.2
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• pp.124-131
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• 2024

Viruses have various applications in the biopharmaceutical industry. They are used in pesticide production, production of vaccines, gene transfers, cancer therapeutics, and more. The downstream processing of viruses is an essential step for their biological and pharmaceutical applications. Among the various processes, the purification of viruses is critical. Membrane chromatography plays a vital role in this process. While ion exchange membrane chromatography is a primarily used method, it has various limitations regarding size exclusion and insufficient purification. Also, it cannot be applied to the rapidly changing strains of viruses such as influenza. This review examines various improved methods of membrane chromatography or alternatives. It focuses on purification, viral recovery rates, and scalability of the methods.

• Journal of KIISE:Software and Applications
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• v.29 no.3
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• pp.133-144
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• 2002

This paper presents a methodology to determine safe persistence of objects from C code during reengineering process. The methodology consists of five steps: the static information methodology, reflection, instantiation, and the refinement. The steps assist to a reengineer to decide appropriate construction and destruction points of an object during its life cycle. Further the steps guarantee safe and consistent interactions among objects.

• Food Science and Preservation
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• v.11 no.4
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• pp.485-490
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• 2004

Crude corn oil (CCO) was obtained through the expression-extraction process from corn germ. The CCOs of final process at $90^{\circ}C,\;70^{\circ}C\;and\;50^{\circ}C$ were stored in outdoor storage tanks. From the samples, refined CCO (RCCO) were prepared with $0\%,\;10\%\;and\;20\%$ excessive of phosphoric acid, caustic solution and acidic clay were used in degumming-alkali refining-bleaching process. RCCOs were stored at room temperature in dark places. The color change was not effected by the amount of phosphoric acid, caustic solution and acidic clay, but temperature of process affects the color change. Finally, the prevention for color reversion of RCCO could be obtained by lowering the temperature of final process and optimal temperature of RCCO in summer was found about less than $50^{\circ}C$.

• Journal of Life Science
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• v.8 no.5
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• pp.598-603
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• 1998

A marine microbe, Vibrio anguillarum was isolated from fish and studied for its concerning pathogenic substance of hemolysin. Purification of hemolysin was achieved by the procedure of ammonium sulfate precipitation from cul-ture filtrate, DEAE-cellulose chromatography, and G-200 gel filtration with 36 fold of purification and 2.3% yield. The molecular weight of the purified hemolysin was 38,000 dalton by SDS-PAGE. The purified hemolysin was stable at pH 6-9, below 45$^{\circ}C$, and up to 1% of NaCl, respectively. $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ inhibited the hemolytic activity whereas EDTA and $Mg^{2+}$ did not.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.223-229
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• 1997

This study was undertaken in order to elucidate the elimination of fenitrothion residues during the dietary fiber and bioflavonoid preparations from mandarin orange peels. Dietary fibers were prepared from contaminated mandarin orange peels through homogenization, enzyme treatment, ethanol precipitation, acetone washing and air drying, at the yields of 17.4% total dietary fiber, 13.1% insoluble dietary fiber and 1.7% soluble dietary fiber. The removal rate of fenitrothion residues at 13 and 0.5 ppm levels in orange peels was 98.4% and 91.9% in total dietary fiber, 99.7% and 97.1% in insoluble dietary fiber, 100% and 99.6% in soluble dietary fiber, respectively. When bioflavonoid was prepared from contaminated mandarin orange peels through homogenization, soaking, ethanol precipitation, hexane and butanol extractions, the removal rate of fenitrothion residues was 92.7% in intermediate extract and 100% in final extract.

• The Korean Journal of Mycology
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• v.20 no.1
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• pp.72-76
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• 1992

The extraction and separation methods of protein-bound polysaccharides from the mycelium and culture broth of Coriolus versicolor (Fr) Quel were investigated. The use of 2% solution of surface active agent Triton X-100, was effective for extraction of the protein-bound polysaccharides from the mycelium. For the separation and partial purification of the protein-bound polysaccharides, the column chromatography using DEAE-Cellulose and DEAE-Sephadex proved to be effective.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.230-236
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• 1993

The isocitrate lyase extracted from Micrococcus luteus was purified 38.8 folds with the overall yield of 10.2%, by the ammonium sulfate fractionation, DEAE-cellulose, 1st Sephadex G-200 and 2nd Sephadex G-200 column chromatography. The purified enzyme showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified enzyme was estimated 60,000 by the SDS-polyacry]amide gel electrophoresis. The apparent Michaelis constant, Km value for isocitrate was 0.95 mM. The optimum pH and temperature of the purified enzyme were pH 7.5 and $40^{\circ}C$, respectively. The enzyme was activated by $Mg^{2+}$ and inhibited by $Mn^{2+}$, $Ca^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $CO^{2+}$. In addition, the activity of isocitrate lyase was increased by glutathione and 2-mercaptocthanol at 5 mM and cysteine at I mM.