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Isolation and Properties of a Cytoplasmic Metalloendoprotease in Escherichia coli (大腸菌 細胞質內 Metalloendoprotease의 抽出과 그 性質에 관하여)

  • Chung, Chin-Ha;Ha, Doo-Bong
    • The Korean Journal of Zoology
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    • v.27 no.4
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    • pp.199-212
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    • 1984
  • A cytoplasmic endoprotease, named protease Ci, has been partially purified by classical chromatographic procedures. This enzyme degrades insulin, glucagon and bovine growth hormone to trichloroacetic acid-soluble materials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. It has a molecular weight of about 120,000 as determined by gel filtration on Sephadex G200, and it appears to be consisted of two identical subunits having molecular weight of 54,000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Protease Ci has an optimum pH of 7.5, and has an isoelectric point of 5.5. This enzyme is a metalloprotease, since it is inhibited by o-phenanthroline and can be activated by the addition of divalent metal cations, such as $Mn^++$ and $Co^++$. Protease ci is inhibited by p-hydroxymercuribenzoic acid, but not by either of leupeptin or Ep475 which are specific inhibitors of sulfhydryl protease. It is distinct from protease Pi, a perplasmic insulin degrading enzyme, since protease Ci is localized to the cytoplasm. The physiological function of protease Ci is presently unknown.

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Electrophoretic analysis of the major proteins of race horse erythrocyte membrane: Their relation to fast erythrocyte sedimentation rate (경주 적혈구막 단백의 전기영동법에 의한 분석 -빠른 적혈구 침강 속도와의 관계-)

  • Bahk, Yeong-woo;Lee, Si-yeong
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.259-264
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    • 1991
  • The proteins of the race horse erythrocyte membrane were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate(SDS-PAGE), and their relations to the fast erythrocyte sedimentation rate(ESR) of the race horse were investigated. The erythrocyte sedimentation rate of race horse were very fast compared with the human one(33 times <$90^{\circ}-plastic-ESR/30m$> and 25 times <$90^{\circ}-micro-ESR/30m$> as fast as the human one) are reported previously. Although the general protein profiles of the race horse erythrocyte membranes were almost similar to that of human, band 3 content was showing higher in race horse (34.7%) than in human (25.3%). The glycoprotein profiles of the race horse erythrocyte membranes revealed by periodic acid Schiff's(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycoprotein) present in human erythrocyte memo brane were almost absent from the Holstein and race horse erythrocyte membranes, but PAS-2 was more in only race horse from that of human. Instead, the bovine erythrocyte membranes showed a strong PAS-B near the origin of the electrophorograms and the race horse erythrocyte membranes showed a strong PAS-negative band near the end of the electrophorograms, which is named as PAS-E in this study. These results suggest that the fast sedimentation rate of race horse erythrocyte is due in part to the presence of more band 3 protein fraction and PAS-E glycoproteins in the race horse erythrocytes.

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Protaetia brevitarsis extract ameliorates the severity of DSS-Induced Ulcerative Colitis in vivo (굼벵이 추출물의 DSS유도 궤양성 대장염 조절효능 탐색)

  • Park, Bog-Im;Jeon, Yong-Deok;Sim, Hyeon-Jae;Lee, Hoon-Yeon;Kim, Seong-Oh;kim, Dong-Keun;Jo, Mi-Na;Cho, Yong-Jin;Kim, Chong-Tai;Kim, Tae-Eun;Jin, Jong-Sik
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.122-122
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    • 2019
  • Protaetia brevitarsis (PB) extracts has been traditionally used as medicinal stuff to treat blood stasis, occlusion of menstruation, tetanus and liver cancer in Asian countries (Korea, Japan, China, Taiwan, India and Myanmar). Especially, Donguibogam, which is traditional korean medicinal book, described the PB extracts as traditional medicine to treat hepatic diseases and vascular disorders. The PB extracts has been considered as highly nutritional food. The major constituents of PB extracts are rich in protein, healthy fats, iron, calcium. Recent studies announced that PB extracts has hepatoprotective effect and anti-microbacterial effect. However, the effect of PB on ulcerative colitis has not been uncovered yet. The aim of this study was to examine the anti-inflammatory effect of PB extracts in dextran sulfate sodium (DSS)-induced colitis mice model. Cytotoxicity of PB was determined by MTT assay and the antiinflammatory effect of PB extract was investigated by measuring nitric oxide (NO) production. PB extracts did not show any cytotoxicity. AIso, PB extracts supressed NO production in LPS-stimulated mice peritoneal macrophages. To determine whether PB could be an effective treatment on ulcerative colitis, DSS was administered in BALB/c mice for 10 days. PB extract significantly improved the clinical signs of DSS-induced UC, including body weight loss, colon length shortening, and disease activity index increase, with histological markers of colon injury. These findings indicated the possibility of PB as a therapeutic agent on ulcerative colitis.

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Cosmeceutical Properties of Fructan (Levan) Produced by Zymomonas mobilis

  • Kim, K. H.;C. S. Han;K. I. Ko;E. K. Yang;Kim, C. H.;Park, S. N.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.700-718
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    • 2003
  • Fructan, a polysaccharide existing in plants or produced by microorganisms, is a sugar polymer of fructose with $\beta$-2,6 linkages. In this study, we investigated some cosmeceutical properties of Fructan such as moisturizing effect, cell proliferation effect, anti-inflammation effect and cell cytotoxicity. Zymomonas mobilis, a microorganism producing Fructan, was cultured in a medium containing 10% sucrose and 2% yeast extract as main components for 24 hours at 37$^{\circ}C$ and pH 7. Fructan was obtained by precipitation from the cultured medium by adding alcohol (alcohol ratio of 1:3) after removing the enzyme by centrifuging. Fructan exhibited almost same moisturizing effect as hyaluronic acid and cell proliferation effect on human fibroblast and keratinocyte as well. Moreover, on cell proliferation test on bio-artificial skin constructed by 3-dimensional(3-D) culture after inducing primary skin inflammation with 0.5% sodium lauryl sulfate (SLS), the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan exhibited higher cell proliferation than the 3-D artificial skin treated with SLS only. On anti-inflammation test on 3-D artificial skin evaluated by measuring secreted quantity of interleukin-1$\alpha$ (IL-1$\alpha$) which is a pre-inflammatory mediator induced by SLS, the quantity of IL-1$\alpha$on the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan was less than the one on the 3-D artificial skin treated with SLS only. As a result of these studies, Fructan has anti-inflammation effect against inflammatory reaction by a skin irritant as well as cell proliferation effect in bio-artificial skin. Fructan was also evaluated as a safe material without any toxicity in safety tests using fibroblasts and animals.

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A study of analytical method for Benzo[a]pyrene in edible oils (식용유지 중 벤조피렌 분석법 비교 연구)

  • Min-Jeong Kim;jun-Young Park;Min-Ju Kim;Eun-Young Jo;Mi-Young Park;Nan-Sook Han;Sook-Nam Hwang
    • Analytical Science and Technology
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    • v.36 no.6
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    • pp.291-299
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    • 2023
  • The benzo[a]pyrene in edible oils is extracted using methods such as Liquid-liquid, soxhlet and ultrasound-assisted extraction. However these extraction methods have significant drawbacks, such as long extraction time and large amount of solvent usage. To overcome these drawbacks, this study attempted to improve the current complex benzo[a]pyrene analysis method by applying the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method that can be analyzed in a simple and short time. The QuEChERS method applied in this study includes extraction of benzo[a]pyrene into n-hexane saturated acetonitrile and n-hexane. After extraction and distribution using magnesium sulfate and sodium chloride, benzo[a]pyrene is analyzed by liquid chromatography with fluorescence detector (LC/FLR). As a result of method validation of the new method, the limit of detection (LOD) and quantification (LOQ) were 0.02 ㎍/kg and 0.05 ㎍/kg, respectively. The calibration curves were constructed using five levels (0.1~10 ㎍/kg) and coefficient (R2) was above 0.99. Mean recovery ratio was ranged from 74.5 to 79.3 % with a relative standard deviation (RSD) between 0.52 to 1.58 %. The accuracy and precision were 72.6~79.4 % and 0.14~7.20 %, respectively. All results satisfied the criteria ranges requested in the Food Safety Evaluation Department guidelines (2016) and AOAC official method of analysis (2023). Therefore, the analysis method presented in this study was a relatively simple pretreatment method compared to the existing analysis method, which reduced the analysis time and solvent use to 92 % and 96 %, respectively.

Geochemistuy of the Borehole Groundwater from Volcanic Rocks in the Northeastern Part of Yeosu Area (여수 북동부 화산암 지역 시추공 지하수의 지화학 특성)

  • 고용권;김경수;배대석;김천수;한경원
    • Economic and Environmental Geology
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    • v.34 no.3
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    • pp.255-269
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    • 2001
  • The geochemical studies on groundwater in the borehole, which is straddled by multi-packer (MP) system, were carried out from a volcanic terrain in the Yeosu area. The pH of groundwater collected from selected sections in the MP-installed borehole is much higher (up to 9.6) than that of the borehole groundwater (7.0-7.9) collected using conventional pumping technique. Hydrochemistry shows that the groundwater has a typical chemical change with increasing sampling depth, suggesting that the groundwater is evolved through water-rock interaction along the fracture-controlled flow paths. The groundwater from the deeper part (138-175 m below the surface) in borehole KI is characterized by the Ca-C11 type with high Ca (up to 160 mg/L) and Cl (up to 293 mg/L) contents, probably reflecting seawater intrusion. The groundwater also has high sodium and sulfate contents compared to the waters from other boreholes. These observed groundwater chemistry is explained by the cation exchange, sulfide oxidation, and mixing process with seawater along the flow path.

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Partial Purification of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju (메주에서 분리된 Enterococcus faecium MJ-14가 생산하는 박테리오신의 부분정제)

  • Lee Jong-Gab;Lee Goon-Ja;Lim Sung-Mee
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.211-216
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    • 2005
  • The bacteriocin produced by E. faecium MJ-14 was precipitated with $50\%$ saturated ammonium sulfate in MRS broth and then the precipitated protein was dissolved in 20 mM sodium phosphate buffer (pH 6.0). The crude bacteriocin was purified by CM-sepharose CL 6B and Sephacry S-100 column chormatograhy. In this case, the purification fold of the bacteriocin was 114, therefore, its activity was 127,293 BU/mg of specific activity. Result from SDS-PAGE of the purified bacteriocin, it was obtained two protein bands of 4.3 kDa and 5.8 kDa having antilisterial activity.

Gingko biloba Extract Ameliorates Colonic Inflammation in DSS-induced Model of Colitis in Mice

  • Rhee, Ki-Jong;Gwon, Sun-Yeong;Hwang, Soonjae;Lee, Chang Gun;Jang, In-Ho;Wie, Myung-Bok;Jung, Bae Dong
    • Biomedical Science Letters
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    • v.20 no.4
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    • pp.227-236
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    • 2014
  • Ulcerative colitis (UC) is a serious gastrointestinal tract disease characterized by recurrent chronic inflammation and mucosal damage of the gastrointestinal tract. The conventional therapies of choice are anti-inflammatory agents, steroids and anti-TNF-${\alpha}$ therapy. However, inherent limitations in these therapies have steered many UC patients to supplement existing therapies with alternative medicinal products. In the current study, we tested the efficacy of Gingko bilola extract (EGb 761) in abating colonic inflammation in a DSS-induced murine model of colitis. C57BL/6 mice were administered 2% DSS in the drinking water for 7 days, then regular water for 7 days, and then 2% DSS for an additional 7 days. EGb 761 (1 mg/dose) was oral gavaged daily for the duration of the experiment. At the termination of the experiment, mice treated with EGb+DSS showed higher body weight, lower spleen weight and longer colon length compared to mice treated with DSS alone. HE-stained colon tissues also exhibited less histologic inflammation in mice treated with EGb+DSS mice compared to mice treated with DSS alone. The serum levels inflammatory cytokines, KC and TNF-${\alpha}$, were also decreased in mice treated with EGb+DSS compared to mice treated with DSS alone. Finally, addition of EGb 761 to TNF-${\alpha}$ treated colonic cell line (HT29/c1) decreased secretion of IL-8 in vitro. These results collectively suggest that EGb 761 abates induction of colitis in DSS-induced model of colitis in mice.

Cathodic Reduction of Dichromate Ion (중크롬산이온의 음극 환원반응)

  • Lee Ju-Seong
    • Journal of the Korean Chemical Society
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    • v.21 no.4
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    • pp.276-283
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    • 1977
  • Reduction of dichromate at a platinum cathode in acid solution was studied by cyclic voltammetry and controlled potential electrolysis. Cathodic polarization curve consisted of three waves in unbuffered solution of potassium dichromate having initial pH ranges 1.5∼4.0, with sodium sulfate as the supporting electrolyte. Relative heights of the first and the second waves were, respectively, a function of chromium (Ⅵ) concentration and activity of hydrogen ion, but that of the third wave was not proportional to both of them. The current of the first two peaks were proportional to the sweep rate of potential (${\nu}$), while that of the last peak vs. ${\nu}^{1/2}$ was linear at the sweep rate of less than 50mV/sec. By the controlled potential electrolysis, the reduction of chromium (Ⅵ) was almost completely suppressed at potentials more negative than the last peak and at initial pH's above ca. 2.3 of unbuffered solution. Therefore, these peaks represented, respectively, $Cr_2O_7^{2-}{\to}Cr^{3+},\;2H^+{\to}H_2$ and the formation of a cathodic film.

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Polymerization of Polystyrene Latex using Ultrasound Energy Effect (초음파 에너지를 이용한 Polystyrene Latex의 중합 특성)

  • Kim, Hyung Jin;Kim, Won II;Lee, Seung Bum;Hong, In Kwon
    • Applied Chemistry for Engineering
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    • v.8 no.6
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    • pp.886-892
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    • 1997
  • There are many methods to synthesize polystyrene latex. Emulsion polymerization technique is commonly used commercially, but it requires a new technology to replace a traditional polymerization method because of the disadvantage of chemical initiator for environmental pollution. Since free radicals can be produced by ultrasound energy effect, polystyrene latex was synthesized using ultrasound energy instead of chemical initiator. As the ultrasonic irradiation time was increased, average molecular weight was increased and polydispersity was decreased. The degree of polymerization was increased with the concentration of SDS and maximum degree of polymerization was shown at 2wt.% SDS concentration and the reaction temperature of $40^{\circ}C$. During the course of polymerization, molecular weight was repeatedly fluctuated because of occurrence of depolymerization. Narrow molecular weight distribution polystyrene latex having controlled molecular weight was synthesized by controlling ultrasonic irradiation time and the concentration of SDS.

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