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Purification and Some Properties of Chitinase from Serratia marcescens JM (Serratia marcescens JM에 의한 Chitinase의 정제와 특성)

  • Lee, Sang Hwan;Yu, Euy-Kyung
    • Journal of the Korean Chemical Society
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    • v.40 no.1
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    • pp.72-80
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    • 1996
  • A chitinase-producing bacterium, Serratia marcescens JM, was isolated from a seashore muds. A chitinase was purified by ammonium sulfate precipitation, affinity adsorption, hydroxylapatite and sephadex G-200 column chromatography. The chitinase obtained from Serratia marcescens JM was purified 42.2 folds with the overall yield of 7.1%. The purified chitinase showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular weight of the enzyme was 59,000 and the apparent kinetic parameters $K_m\;and\;V_{max}$ for the purified chitinase were 5.17 mg/mL and 39.8 unit/mL, respectively. The optimum pH and temperature of the purified chitinase were 7.0 and 50$^{\circ}C$, respectively and the purified enzyme was stable on pH 7.0 up to 50$^{\circ}C$. The enzyme were activated by $Cu^{2+},\;Ca^{2+}\;and\;Mg^{2+}$ and inhibited by $Hg^{2+}$ respectively. In addition, Cysteine increased the chitinase activity and EDTA, MIA, PCMB and SDS inhibited enzyme activities. Major cations, $MG^{2+},\;Ca^{2+},\;K^+\;and\; Na^+$ present in seawater slightly stimulated the chitinase activity.

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Purification of Hemolysin from Vibrio anguillarum Isolated from Fish (어류분리 Vibrio anguillarum 용혈소의 정제)

  • 김영희
    • 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.

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Electrochemical synthesis of nanosized hydroxyapatite/graphene composite powder

  • Vesna, Miskovic-Stankovic;Sanja, Erakovic;Ana, Jankovic;Maja, Vukasinovic-Sekulic;Miodrag, Mitric;Jung, Young Chan;Park, Soo Jin;Rhee, Kyong Yop
    • Carbon letters
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    • v.16 no.4
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    • pp.233-240
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    • 2015
  • Electrochemical synthesis was employed to prepare a novel hydroxyapatite/graphene (HAP/Gr) composite powder suitable for medical applications as a hard tissue implant (scaffold). The synthesis was performed in a homogeneous dispersion containing Na2H2EDTA·2H2O, NaH2PO4 and CaCl2 with a Ca/EDTA/PO43− concentration ratio of 0.25/0.25/0.15M, along with 0.01 wt% added graphene nanosheets, at a current density of 137 mA cm−2 and pH value of 9.0. The field emission scanning electron microscopy and transmission electron microscopy observations of the composite HAP/Gr powder indicated that nanosized hydroxyapatite particles were uniformly placed in the graphene overlay. Raman spectroscopy, Fourier transform infrared spectroscopy and X-ray diffraction confirmed graphene incorporation in the HAP/Gr powder. The electrochemically prepared HAP/Gr composite powder exhibited slight antibacterial effect against the growth of the bacterial strain Staphylococcus aureus.

[$Zn^{2+}$ Modulates the Responses of Rat Dorsal Horn Neuron to C-Fiber Stimulation and Excitatory Amino Acids

  • Ahn, Chang-Hoon;Shin, Hong-Kee;Kim, Jin-Hyuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.6
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    • pp.455-461
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    • 2000
  • Zinc contained in the neurons of central nervous system is activity-dependently released and then attenuates NMDA (N-methyl-D-aspartate)-induced neurotoxicity while augmenting non-NMDA-induced neurodegeneration. Zinc also has been reported to produce antinociceptive action on the inflammation- and nerve injury-induced hyperalgesia in the behavioral test. In this study, we investigated the effects of zinc on the responses of dorsal horn cells to NMDA, kainate and graded electrical stimulation of C-fibers. In the majority of WDR cells (70.6%), zinc current-dependently inhibited WDR cell responses to NMDA and in the remaining cells, produced biphasic responses; excitation followed by inhibition. Zinc augmented the responses of WDR cells to iontophoretical application of kainate. The dominant effect of $Zn^{2+}$ on the responses of WDR cells to C-fiber stimulation was excitatory, but inhibition, excitation-inhibition and no change of the responses to C-fiber stimulation were induced. $Ca^{2+}-EDTA$ antagonized the excitatory or inhibitory effects of $Zn^{2+}$ on the WDR cell responses. These experimental findings suggest that $Zn^{2+}$ modulates the transmission of sensory information in the rat spinal cord.

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Statistical Optimization of Medium Components for the Production of Biosurfactant by Bacillus licheniformis K51

  • Joshi Joshi;Sanket Sanket;Yadav Sanjay;Nerurkar Anuradha;Desai Anjana J.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.313-319
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    • 2007
  • The nutritional medium requirement for biosurfactant production by Bacillus licheniformis K51 was optimized. The important medium components, identified by the initial screening method of Plackett-Burman, were $H_3PO_4,\;CaCl_2,H_3BO_3$, and Na-EDTA. Box-Behnken response surface methodology was applied to further optimize biosurfactant production. The optimal concentrations for higher production of biosurfactants were (g/l): glucose, $1.1;NaNO_3,\;4.4;MgSO_4{\cdot}7H_2O,\;0.8;KCl,\;0.4;CaCl_2,\;0.27;H_3PO_4,\;1.0ml/l;\;and\;trace elements\;(mg/l):H_3BO_3,\;0.25;CuSO_4,\;0.6;MnSO_4,\;2.2;Na_{2}MoO_4,\;0.5;ZnSO_4,\;6.0;FeSO_4,\;8.0;CoCL_2,\;1.0;$ and Na-EDTA, 30.0. Using this statistical optimization method, the relative biosurfactant yield as critical micelle dilution (CMD) was increased from $10{\times}\;to\;105{\times}$, which is ten times higher than the non-optimized rich medium.

Studies on the Myofibrilar Protein from Korean Duck Muscle (오리고기의 근원섬유 단백질에 관한 연구)

  • Chang, In-Yae;Nam, Hyun-Keun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.9 no.1
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    • pp.45-50
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    • 1980
  • Myofibrilar Protein from Korean Duck Muscle was extracted and ATPase activities were studied. The results were as follows: 1. Mg-activated ATPase activity of Myofibril from Korean Duck, muscle exhibited a biphasic response, ATPase activity was high at a low ionic strength and low activity was showed at high ionic strength. 2. Effect of EDTA on the Myofibrillar protein ATPase activity was studied, it was investigated that the EDTA inhibition was showed at the concentration of $6.9{\mu}g$ and it above. 3. It showed that the effect of Ca++ on ATPase activity was decreased at the lower than $3{\mu}g$. Inhibition showed at the concentration of $6.9{\mu}g$ and it above. 4. It showed that the effect of Mg++ on ATPase activity was decreased at the lower than $3.6{\mu}g$. Inhibition showed at the concentration of $3.9{\mu}g$ and it above.

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a Study on the Optimal Location Evaluation of Airport Terminal Facilities combinded the Accessibility Theory and Spatial Analysis Model of GIS in Seoul Metropolitan Area (접근성이론과 GIS공간분석기법의 접목을 통한 도시시설의 최적입지 평가방법 연구 - 수도권 도심공항터미널 입지후보지 평가를 중심으로 -)

  • Kim, Hwang-Bae;Kim, Si-Gon
    • 한국공간정보시스템학회:학술대회논문집
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    • 2000.06a
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    • pp.189-201
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    • 2000
  • 접근성이란 특정 목적을 가진 통행자가 한 지점에서 다른 지점으로 얼마나 쉽게 갈 수 있는가 를 나타내는 척도를 말한다. 따라서 접근성은 주로 공간적인 거리나 시간이 가장 중요한 요인으로 작용하며 최근에는 공간적 거리보다는 접근시간이 더 중요한 접근성 척도로 사용되고 있다. 본 연구는 GIS공간분석기법과 접근성이론을 접목하여 도시시설의 적정후보지 평가방법을 정립한 후 현재 검토되고 있는 수도권 도심공항터미널 입지 후보지 중 어떤 후보지가 이용들의 총통행시간을 최소화하는 후보 지인가에 대해 평가해보았다. 평가 결과 다음과 같은 사실을 밝힐 수 있었다. 첫째, 수도권 전체로 볼 때 평균접근시간이 가장 양호한 후보지는 쌍문터미널(현 간이 도심공항터미널)이고, 2위는 현 삼성동 도심공항터미널과 강남고속터미널, 3위는 동서울터미널과 남서울터미널 순으로 나타났다. 둘째, 수도권전체 이용자들의 총통행시간을 최소로 하는 도심공항터미널 후보지는 접근성이 가장 양호한 쌍문동터미널이 아니라 현 서울역이 1위이고, 2위는 강남고속 터미널, 3위는 용산 역으로 나타났다. 셋째, 터미널 후보지간의 가중평균접근 시간은 도심인 서울시내에 입지한 후보지와 수도권 외곽에 입지 한 후보지간에 큰 차이가 없으나 총 이용자 접근시간은 서울시에 입지한 후보지보다 외곽의 후보지가 훨씬 높게 나타나 뚜렷한 차별성을 보이고 있다. 넷째, 최적후보지 1,2,3순위 모두 서울도심과 강남도심에 입지한 지역들로 나타나 교통의 접근성 보다는 아직 인구밀집도가 주요 도시시설의 입지결정에 주요한 결정요인 되는 것으로 나타났다.해석 시스템을 구축할 예정이다. 추후에는 하수도 관망해석 컴포넌트와 하수도 업무 컴포넌트와의 통합부분에 대한 연구가 진행되어야 할 것이다.7.0로 하고 표준(標準) EDTA 용액(溶液)을 소량(少量)넣고 8N-KOH로 pH $12{\sim}13$으로 하고 N-N 희석분말(稀釋粉末)을 지시약(指示藥) 으로써 표준(標準) EDTA 용액(溶液)으로 적정(滴定)하여 Ca 치(値)를 얻었다. Ca와 Mg의 합계결정치(合計決定値)와 Ca 적정치(滴定値) 차(差)로 Mg 치(値)를 얻었다. 음(陰) ion 구분(區分)으로부터 상법(常法)에 의하여 $MgNH_4PO_4$의 침전(沈澱)을 만들어서 HCl에 녹키고 일정량(一定量)의 표준(標準) EDTA 용액(溶液)을 넣어 pH 7.0로 한다음 완충액(緩衝液)으로 pH 10으로 하고 BT 지시약(指示藥)을 써서 표준(標準) Mg $SO_4$용액(溶液)으로 적정(滴定)하여 P 치(値)를 얻었다. 본법(本法)으로 Na-phytate를 분석(分析)한 결과(結果) Na-phytate의 분자식(分子式)을 $C_6H_6O_{24}P_6Mg_4CaNa_2{\cdot}5H_2O$라고 하였을 때의 이론치(理論値)에 비(比)하여 P가 98.9% Cark 97.1%, Mg가 99.1%이고 통계처리(統計處理)한 결과분석치(結果分析値)와 이론치(理論値)는 잘 일치(一致)된다. 그러나 종래법(從來法)에 의(依)한 분석치(分析値)는 이론치(理論値)에 비(比)하여 P가 92.40%, Cark 86.80

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Effect of microwave heating and salts addition on pH and acidity of Kakdugi during fermentation (깍뚜기 발효중 순간 가열과 염첨가가 pH변화에 미치는 영향)

  • Yun, Jung-Won;Kim, Jong-Koon;Lee, Jung-Kun;Kim, Woo-Jung
    • Applied Biological Chemistry
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    • v.34 no.3
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    • pp.213-218
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    • 1991
  • Addition of salts or their mixtures and microwave heating were studies for their effects on Kakdugi fermentation. The Kakdugi, a Korean fermented Chinese radish, were prepared by salting in 15% NaCl solution and fermented at $25^{\circ}C$. From the results, a first order reaction relationship was found between salt permeated into the radish and time during brining in $5{\sim}10%$ NaCl solution. Addition of 0.05M KCl into 15% NaCl brining solution or microwave heating of salted radish for 3 minutes showed a little decreasing effect on Kakdugi fermentation rate while beating for 1 or 2 minuted resulted a rather increase. When three different salt mixtures in the concentration range of $0.001{\sim}0.01\;M$ were added into half fermented Kakdugi(appr. pH 4.4), the fermentation was greatly controlled based on pH change. Among the salt mixtures, mixture III$(Na_2HPO_4,\;Na_2PO_4,\;NaNO_2,\;Ca\;EDTA,\;Sod.\;citrate)$ showed a most significant effect where the time required to reach pH 4.0 after addition was extended by more than 6 folds when it was compared to the control method.

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Yarrowia lipolytica TH65가 생산하는 Alkaline Proteinase의 정제 및 특성

  • Yu, Choon-Bal;Kim, Chang-Hwa;Jin, Young-Ho;Jin, Ing-Nyol
    • Microbiology and Biotechnology Letters
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    • v.24 no.3
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    • pp.316-320
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    • 1996
  • An alkaline proteinase produced by Yarrowia lipolytica TH65 was purified by 40-65% ammonium sulfate fractionation, DEAE-cellulose chromatography, and gel filtration with Sephadex G-100 and Sephadex G-75. The purified enzyme was shown as a single band on SDS-PAGE, and its molecular weight 31,500. Optimum temperature and pH were 40$\circ$C and 8.5-9.0, respectively, and the enzyme was stable below 40$\circ$C and in the pH range of 6-8. The enzyme was strongly inhibited by divalent ions, completely by PMSF, and partially by EDTA, EGTA, and phenanthroline. But the inhibitory effect in the presence of EDTA, EGTA and phenanthroline could be reversed by addition of Ca$^{2+}$. Thus, these results indicated that the purified enzyme was an alkaline serine proteinase (E.C. 3.4.21.14).

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Metallo-collagenase production by Arthrobacter creatinolyticus KP015744

  • Savita A. Kate;Madhuri Sahasrabudhe;Archana Pethe
    • Advances in environmental research
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    • v.11 no.1
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    • pp.1-16
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    • 2022
  • Amongst 27 isolates from deteriorated leather samples, Arthrobacter creatinolyticus KP015744 zzx28 was found to be an efficient collagenase producer. Collagenase production of 13.33 µmoles/min was shown at an optimum temperature at 37℃ after 72h and at pH 7.5 by using 2 ml/dL inoculum in 10 mg/ml collagen peptide type I as a substrate. In presence of Hg2+, EDTA and 𝛽-mercaptoethanol the collagenase production by the isolate was strongly inhibited however Fe2+, Ca2+and DMSO enhanced production of the enzyme. Specific activity was found to be 19.46×103 U/mg and molecular weight 66 kD by SDS PAGE. Isolate also has potential to hydrolyze keratin which is another important protein found in leather. Experimental results propose that collagenase can be effectively used as a tool for collagen and keratin rich solid waste treatment.