• Title/Summary/Keyword: Gelatin (Gel)

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Purification and Characterization of the S-type Pyocin of Pseudomonas aeruginwa 90-2-2205 Isolated from Korean Patients (한국환자유래의 녹농균 90-2-2205로부터 S형 Pyocin의 정제 및 특성)

  • 김란숙;이정미;박영덕;진익렬;김병오
    • Microbiology and Biotechnology Letters
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    • v.21 no.2
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    • pp.132-138
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    • 1993
  • The s-type pyocin was purified from the lysate of the mitomycin C-induced Pseudomonas aeruginosa 90-2-2205 cells by the other of ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Sephadex G-200 gel filtration. The purity was confirmed by the polyacry-lamide gel electrophoresis. The molecular weight of the purified pyocin was estimated 180, 000 by gel filtration. The pyocin was analyzed to be a complex of some polypeptides by the SDS-PAGE. The pyocin was stable by heat treatment and at pH 6-7.5 by adding 10-3% gelatin and 0.2M NaCl to the 10mM Tris-HCl buffer (pH7.5). Its killing action against the sensitive cells was assumably a single hit process.

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Changes in Ice Dendrite Size during Freezing Process in Gelatin Matrix as a Model Food System (모델 식품으로 젤라틴 매트릭스에서 동결과정에 따른 얼음 결정체 변화)

  • Min, Sang-Gi;Hong, Geun-Pyo;Choi, Mi-Jung
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.312-318
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    • 2008
  • The objective of this study was to investigate the changes in ice dendrite size during freezing process in gelatin matrix as a model food system in order to provide mathematical relation between freezing condition and ice dendrite size. Gelatin gel as a model matrix was frozen in unidirectional Neumann's type of heat transfer. The thermograms' analysis allowed to determine the freezing temperature of the sample, the position of the freezing front versus time, and thus, freezing front rate. The morphology of ice dendrites was observed by scanning electron microscopy after freeze-drying. We observed that the means size of ice dendrite increased with the distance to the cooling plate; however, it decreased with the cooling rate and the cooling temperature. In addition, the shorter durations of the freeze-drying process was shorter decreeing the decreased the freezing front rate, resulted in their resulting in a larger pore size of the ice dendrite pores for the sublimation channel of that operate as water vapor sublimation channels. From these results, we could derive a linear regression as an empirical mathematical model equation between the ice dendrite size and the inverse of freezing front rate.

Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography (젤라틴 친화성 크로마토그래피를 이용한 스파르가눔 성분단백질의 순수분리)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.1-8
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    • 1991
  • Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum) , 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

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Screening for raw material of modified gelatin in marine animal skins caught in coastal offshore water in Korea (수식 어류껍질 젤라틴의 원료로서 연근해산 수산물껍질의 검색)

  • Cho, Soon-Yeong;Kim, Jin-Soo
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.134-139
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    • 1996
  • In order to effectively utilize marine animal skin wastes in marine processing manufacture, conger eel skin, file fish skin and arrow squid skin as raw material of edible gelatin were screened. Conger eel skin was the highest in the collagen content, followed by Ole fish skin and arrow squid skin, in the order named. In the fish skins, the soluble and insoluble collagens occupied $67.4%{\sim}72.3%\;and\;27.7{\sim}32.6%$, respectively, and in the arrow squid skin, 30.4ft and 69.6ft, respectively. No difference in the amino acid composition between soluble and insoluble collagens was detected. Collagen from the marine animal skin catched in coasted and offshore water in Korea consisted ${\alpha}$ chain and ${\beta}$ chain, and ${\alpha}$ chain were hetero type. The sum of proline and hydroxyproline contents in conger eel skin collagen was higher than that in the other skin collagens, while was lower than that pork skin collagen. Conger eel skin collagen exhibited a higher denaturation temperature in solution and a higher degree of proline hydroxylation, compared with skin collagen of the respective species. The physical properties such as gel strength, melting point and gelling point of conger eel skin gelatin were superior to those of file fish skin and arrow squid skin gelatins.

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Estimation of the Characteristics for the Dose Distribution in the Polymer Gel by Means of Monte Carlo Simulation (몬테카를로 시뮬레이션을 이용한 양성자 조사에 따른 Polymer Gel 내부의 선량 분포 특성 평가)

  • Park, Min-Seok;Kim, Gi-Sub;Jung, Hai-Jo;Park, Se-Young;Choi, In-Seok;Kim, Hyun-Ji;Yoon, Yong-Su;Kim, Jung-Min
    • Journal of radiological science and technology
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    • v.36 no.2
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    • pp.165-173
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    • 2013
  • This study was the estimation of the dose distribution for proton, prompt gamma rays and proton induced neutron particles, in case of exposing the proton beam to polymer gel dosimeter and water phantom. The polymer gel dosimeter was compositeness material of Gelatin, Methacrylic acid, Hydroquinone, Tetrakis and Distilled water. The density of gel dosimeter was $1.04g/cm^3$ which was similar to water. The 72, 116 and 140 MeV proton beams were used in the simulation. Proton beam interacted with the nuclei of the phantom and the nuclei in excited states emitted prompt gamma rays and proton induced neutron particles during the process of de-excitation. The proton particles, prompt gamma rays, proton induced neutron particles were detected by polymer gel dosimeter and water phantom, respectively. The gap of the axis for gel was 2 mm. The Bragg-peak for proton particles in gel dosimeter was similar to water phantom. The dose distribution for proton and prompt gamma rays in gel dosimeter and water phantom was approximately identical in case of 72, 116 and 140 MeV for proton beam. However, in case of proton induced neutron particles for 72, 116 and 140 MeV proton beam, particles were not detected in gel dosimeter, while the Water phantom absorbed neutron particles. Considering the resulting data, gel dosimeter which was developed in the normoxic state attentively detected the dose distribution for proton beam exposure except proton induced neutron particles.

Nano-scale Proteomics Approach Using Two-dimensional Fibrin Zymography Combined with Fluorescent SYPRO Ruby Dye

  • Choi, Nack-Shick;Yoo, Ki-Hyun;Yoon, Kab-Seog;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • v.37 no.3
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    • pp.298-303
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    • 2004
  • In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

Raman Spectroscopy of Irradiated Normoxic Polymethacrylic Acid Gel Dosimeter

  • Bong, Ji-Hye;Choi, Kyu-Seok;Yu, Soo-Chang;Kwon, Soo-Il;Cho, Yu-Ra;Park, Chae-Hee;Park, Hyung-Wook
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.625-629
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    • 2011
  • A quantitative analysis of the decreasing rate of the monomer and increasing rate of the polymerization was made by monitoring radiation level increments using Raman spectroscopy within the therapeutic radiation range for a normoxic polymethacrylic acid gel dosimeter. The gel dosimeter was synthesized by stirring materials such as gelatin, distilled water, methacrylic acid, hydroquinone and tetrakis phosphonium chloride at $50^{\circ}C$, and the synthesized gel was contained in a 10- mm diameter and 32-mm high vial to conduct measurement. 24 hours after gel synthesis, it was irradiated from 0 Gy to 20 Gy by 2 Gy using a Co-60 radiotherapy unit. With use of the Cryo FE-SEM, structural changes in the 0 Gy and 10 Gy gel dosimeters were investigated. The Raman spectra were acquired using 532-nm laser as the excitation source. In accordance with fitting the changes in C-COOH stretching (801 $cm^{-1}$), C=C stretching (1639 $cm^{-1}$) and vinyl $CH_2$ stretching (3114 $cm^{-1}$) vibrational modes for monomer and $CH_2$ bending vibrational mode (1451 $cm^{-1}$) for polymer, sensitive parameter S for each mode was calculated. The values of S for monomer bands and polymer band were ranged in $6.0{\pm}2.6$ Gy and $7.2{\pm}2.3$ Gy, respectively, which shows a relatively good conformity of the decreasing rate of monomer and the increasing rate of polymerization within the range of error.

Formulation of Water-soluble Topical Preparations of Epidermal Growth Factor (상피세포성장인자의 수용성 외용제제화)

  • Lee, Yoo-Cheol;Park, Eun-Seok;Chi, Sang-Cheol
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.177-184
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    • 1995
  • In order to formulate an aqueous topical preparation of epidermal growth factor(EGF) for the treatment of open wound and bum, the stability of EGF in aqueous vehicles containing various stabilizers was evaluated and the pharmacological activity of gel preparations formulated with poloxamer 407 was determined with wound model. Various additives, which are known as potent stabilizers for proteins and polypeptides so far, were used to increase the stability of EGF in aqueous vehicles. The contents of EGF in the vehicles containing stabilizers were determined with an HPLC method after the storage at $37^{\circ}C$. EGF was more stable in ultrapure water than RO water or saline. All the additives studied resulted in deleterious effects on EGF stability. Therefore, it was speculated that any additives or impurities in the vehicle made EGF unstable. However, nitrogen purge of solution increased the stability of EGF in aqueous vehicles. The aqueous topical preparations of EGF were formulated with poloxamer 407 as a gel base in saline. Gelatin or amastatin was employed as a protease inhibitor. The pharmacological effect of EGF gel was studied with open wound model in mice. EGF preparations, made of oleaginous base or poloxamer gel base, showed significant healing effect compared to the control group(p<0.05). The addition of protease inhibitor in poloxamer 407 gel resulted in significant healing effect compared to the gel without it(p<0.05). Body weights of mice treated with EGF preparation were increased at the first day after the formation of open wound, while those of the control group were decreased. The EGF gel made of poloxamer 407 containing a pretense inhibitor would be a promising aqueous topical preparation for EGF.

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Characterization of extracellular proteases from alkalophilic vibrio sp. strain RH 530

  • Kwon, Yong-Tae;Moon, Sun-Young;Kim, Jin-Oh;Kho, Yung-Hee;Rho, Hyune-Mo
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.501-506
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    • 1992
  • An alkalophilic Vibrio sp. RH530 showing high proteolytic activity was isolated form soil samples by enrichment culture. The activity staining using gelatin SDS- polyacrylamide gel electrophoresis (PAGE ) revealed that the strain produced an alkaline major protease (Apr B) with a size of 27 kDa, and at least six minor proteases. The apparent sizes of four of the minor proteases were approximately 45, 28, 22 and 19 kDa. Apr B and five of the minor proteases were inhibited by serine protease inhibitors including PMSF and DFP, suggesting that they are serine proteases. One of the minor proteases was inhibited by metalloprotease inhibitors, not by serine protease inhibitors, indicating it to be a metalloprotease. Furthermore, the activities of Apr B and Prt 3 were not inhibited by SDS in the reaction mixture. The production of Apr B and some of the minor proteases was specifically affected by culture temperature (30 to 37.deg.C) and pH (7 to 10). The production of Apr B. Prt 2, Prt 5 and Prt 6 was mainly affected by culture temperature, while Prt 4 by culture pH. Prt 1 and Prt 3 were not affected by neither of these factors.

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Characterization of Plasmid DNA in Streptococcus faecalis var. liquefaciens (Streptococcus facalis var. liquefaciens에 존재하는 Plasmid DNA의 특성)

  • 강국희;이명기;박연희
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.417-422
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    • 1985
  • Streptococcus faecalis var. liquefaciens was examined for the presence of plasmid deoxyribonucleic acid. An analysis by agarose gel electrophoresis revealed the presence of at least four plasmids of approxymately 6.8, 5.2, 2.6, and 2.1 Mdal. Two plasmid cured strains were obtained by novobiocin treatment. SKR2, which lost 5.2 mdal plasmid (pSK2) and 2.1 Mdal plasmid(pSK4) was sensitive to lincomycin and erythromycin. However, all cured strains showed identical response as parental strain in sugar fermentation, temperature sensitivity, proteolytic activity, and liquefaction of gelatin. The results imply that pSK2 or pSK4 is associated with antibiotic resistance of Str. faecalis var. liquefaciens.

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