• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.230.3-231
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• 2003

The therapeutic use of protein pharmaceuticals produced by recombinant DNA technology is increasing in recent decades. In order to investigate the quality of recombinant proteins, it is important to identify and assign the impurities produced in the process of recombination or in storage conditions. Capillary Electrophoresis is emerging technology exhibiting high sensitivity, selectivity and speed and may be most powerful tools for this application. In this study, human growth hormone (hGH) has been analyzed by various mode of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary gel electrophoresis (CGE), and capillary isoelectric focusing (cIEF) to indicate the chemically or biologically oriented variants and the degraded fragments. (omitted)

• 한국버섯학회지
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• 제19권4호
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• pp.316-321
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• 2021

리그닌 분해효소의 생산은 나선형 리본이 있는 새로운 형태의 회전식 통풍관 생물 반응기(RTB)를 사용하여 느타리(Pleurotus ostreatus) No.42에 의해 실시하였다. 락게이즈(laccase)의 최대 생산량은 배양 3일 후 약 8,200 U/bioreactor 수준에 도달한 후 감소하였다. 반면에, 망간퍼옥시데이즈(MnP)의 최대 생산은 6일 배양 후 약 8,400 U/bioreactor의 수준에 도달하였다. 그러나 이 발효조에서 리그닌퍼옥시데이즈(LiP)는 검출되지 않았다. 그 결과 회전식 통풍관 생물 반응기(RTB)가 리그닌 분해효소를 대규모 생산을 위해 성공적으로 생산할 수 있음을 보여주었다. 이 발효기에서 망간퍼옥시데이즈의 정제 과정은 Sepharose CL-6B, Superdex 75 prep grade 및 Mono-Q에 대한 크로마토그래피를 포함하여 정제하였다. 이 주요 효소는 sodium dodecyl sulfate-polyacrylamide겔 전기영동(SDS-PAGE)에서 분자량 36,400, pI 3.95의 등전점(IEF)으로 각각 확인되었다. 이 발효기의 주요 효소 N-말단 서열은 정치 및 진탕배양과 같은 다른 배양조건에서 보고된 MnP3 효소와 동일하였다.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.25-31
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• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.607-611
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• 2004

Glycinin, one of the predominant storage proteins in soybeans, was examined as to whether it could be used as a calcium-binding mediator after chemical phosphorylation and enzymatic hydrolysis. Glycinin is composed of six subunits. One of the proglycinin subunits $(A_{la}B_{lb})$ was overexpressed in E. coli to obtain nonphosphorylated proteins with homogeneity. To investigate the enhanced calcium-binding properties of the phosphopeptides, the proglycinin was purified, phosphorylated, and hydrolyzed with trypsin. The proglycinin expressed in E. coli was purified by ammonium sulfate precipitation, ion-exchange chromatography, and cryoprecipitation. Chemical phosphorylation by sodium trimetaphosphate was performed to obtain phosphorylated proglycinin. After the phosphorylation, one-dimensional isoelectric focusing gel electroanalysis confirmed the phosphorylation of the proglycinin. The phosphorylated peptides were then hydrolyzed with trypsin, followed by a binding reaction with calcium chloride. The calcium-bound phosphopeptides were finally separated using immobilized metal $(Ca^{2+})$ chromatography. Consequently, a limited tryptic hydrolysate of the isolated phosphopeptides exhibited an enhanced calcium-binding ability, suggesting the potential of glycinin phosphopeptides as a calcium-binding mediator with greater availability.

• The Korean Journal of Ecology
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• 제23권2호
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• pp.187-192
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• 2000

After Cassia mimosoides var. nomame was treated with Phytolacca americana extracts during the germination process, its effect on the activities of AsPOX and GuPOX were examined. As the concentration of P. americana extract increased. the activity of AsPOX decreased while that of GuPOX increased. During the germination process, the activity of AsPOX was lower than the control, while the activity of GuPOX was over 2 times higher than the control. Activity of total peroxidase by IEF was extremely high at pI 6.4 and pI 6.6 when treatment was made with a 30% concentration of P. americana extract. The treatment with phenolic compounds, caffeic acid and benzoic acid did not show much difference from the control although a slight increase was observed at pI 6.6. Activity of GuPOX in C. mimosoides was over 4 times higher in roots than in shoots. Namely, because GuPOX activity of C. mimosoides was increased by extracts of P. americana, defense enzyme, GuPOX. was generated against external stress, and we could certified the activity increase at pI 6.4, especially in root.

• 대한의생명과학회지
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• 제12권3호
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• pp.177-183
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• 2006

To demonstrate the effect of formulation conditions and evaluations of structural integrity from ovalbumin containing poly lactide glycolide copolymer (PLGA) microspheres for Vaccine delivery, OVA microspheres were prepared by a W/O/W multiple emulsion solvent extraction technique. Dichloromethan (DCM) and Ethyl acetate (EA) were applied as an organic phase and poly vinyl alcohol (PVA) as a secondary emulsion stabilizer. Microspheres were characterized for particle size, morphology (optical microscopy and Scanning Electron Microscope (SEM)). Protein denaturation was evaluated by size exclusion chromatography (SEC), SDS-PAGE and isoelectric focusing (IEF). Residual organic solvent was estimated by gas chromatography (GC) and differential scanning calorimetry (DSC). Optical photomicrograph and SEM revealed that micro spheres were typically spherical but various morphologies were observed. Mean particle size $(d_{vs})$ of microspheres were in the range of $3{\sim}50{\mu}m$. Also, The protein stability was not affected by the fonnulation process and residual organic solvent was beyond the detection below 0.1ppm. These results demonstrated that micro spheres might be a good candidate for the parenteral vaccine delivery system.

• BMB Reports
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• 제36권2호
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• pp.214-222
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• 2003

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

• 혜화의학회지
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• 제13권1호
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• pp.97-105
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• 2004

In the Oriental medicine, it was recognized that mind and body have a inseparable relationship. The goal of this study was to examine the body(肉體) and self-be1ief(自己信念) and methods of self-therapy(自己治癒) in transcendental meditation(冥想療法). We will furnish the information of the basic concepts and methods in transcendental meditation. The findings are as follows: 1. The thing that you have in mind does not stay in abstract idea but materializes and operates the body. 2. When you have disease implies that you have negative belief, thought and mind. 3. When you can observe the things happening in your body and mind- just like meditation- you can have positive changes. 4. When you change the negative beliefs into positive ones on body, you feel gratitude and the disease will be cured.

• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.259-266
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• 1996

형태학적 제3군 가시아메바의 taxonomic validity는 아직 확실하지 않다. 이번 연구에서 제3군에 속하는 6종의 가시 아메바 즉 A. culbertsoni A. healyi A. palestinensis. A. pustulosa, A. royreba 및 A. lenticulata의 type strain들의 동위효소. 미토콘트리아 DNA 및 small subunit(ssu) rDNA의 Restriction Fragment Length Polymorphism(RFLP) 양상을 비교하여 이들의 taxonomic validity를 검토하였다. 미토콘드리아 DNA의 RFLP 양상은 분리주간에 서로 심한 차이를 보였다. A. palestinensis와 A. pustulosc는 거의 동일한 rDNA RFLP(추정 염기 치환율 2.6%) 및 동위효소의 양상을 보여 A. palestinensis와 A. pustulosc는 같은 종으로 판단되었다. 그외의 종들은 서로 아주 다양한 rDNA RFLP 및 동위효소의 양상을 나타내어 독립종으로 인정되었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제16권2호
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• pp.37-48
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• 2008

The diapause and non diapause associated proteins of multivoltine silkworm eggs were analysed by two dimensional (2D) gel electrophoresis. The study was made at 0 hr, 24 hrs and 48 hrs after oviposition. A total of four protein spots in diapause eggs at 24 hrs of oviposition and two protein spots in non diapause eggs at 0 hrs of oviposition were observed. All the six protein spots were considered to have association with diapause and non diapause characters. The molecular weight (MW) and isoelectric point (PI) of these 6 protein spots were calculated. The protein spots 1 and 2 observed in 0 hr of non diapause eggs were found to have the MW of 67 and 75 KDa and PI of 8.6 and 8.4 respectively. Similarly the four protein spots observed in diapause egg at 24 hrs of oviposition exhibited MW viz., 15, 17,20 and 25 KDa and PI of 5.3, 5.8, 6.5 and 6.0 respectively. All these 6 identified protein spots were subjected to in-gel digestion and resulted tryptic peptides were analyzed by Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF-MS). Databases searched based on experimentally determined molecular weights of peptides for the determination of the identities of proteins. The identified proteins indicated homology of 34% to 95%. The results indicate that the proteins may playa role in development of diapause and non diapause eggs.