• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.328-335
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• 1998

Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.109-109
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• 2018

Bacillus subtilis B22, a chemotrophic and aerobic bacterial strain was isolated from homemade kimchi, identified by 16S rRNA gene sequencing. B22 was primarily screened by biochemical, carbon source utilization tests. B22 was used to produce pectinase and ${\beta}$-glucosidase by submerged fermentation under different light sources. B22 was incubated in pectin media and basal media (pH 7.0) under blue, green, red and white light-emitting diodes (LEDs), fluorescent white light, and in darkness at $37^{\circ}C$, orbital shaker 150 rpm for 24 hours. Fermentation under blue LEDs maximized pectinase production ($71.59{\pm}1.6U/mL$ at 24 h) and ${\beta}$-glucosidase production ($56.31{\pm}1.6U/mL$ at 24 h). Further, the production of enzyme increased to pectinase ($156{\pm}1.28U/mL$) and ${\beta}$-glucosidase ($172{\pm}1.28U/mL$) with 3% glucose as a carbon source. Activity and stability of the partially purified enzymes were higher at pH 6.0 to 8.0 and $25-55^{\circ}C$. The effect on the metal ions $Na^+$ and $K^+$ and (moderateactivity) $Mn^{2+}$ and $Ni^{2+}$ increased activity, while $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, and $Fe^{2+}$ inhibited activity. EDTA, phenylmethylsulfonyl fluoride and 5,5-dithiobis (2-nitrobenzoicacid) reduced activity, while tetrafluoroethylene and 1,10-phenanthroline inhibited activity. The amylase was highly tolerant of the surfactants TritonX-100, Tween-20, Tween-80 and compatible with organic solvents methanol, ethanol, isoamylalcohol, isopropanol, t-butylalcohol and the oxidizing agents hydrogen peroxide, sodium perborate and sodium hypochlorite, although potassium iodide and ammonium persulfate reduced activity. These properties suggest utility of pectinase and ${\beta}$-glucosidase produced by B. subtilis B22 under blue LED-mediated fermentation for industrial applications.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.78-83
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• 1994

Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneous state fron an acetic acid producing bacteria, Acetobacter sp. HA. The enzyme was purified about 153-fold with an overall yield of 35% from the crude cell extract by solubilization and extraction of the enzyme with Triton X-100 and subsequent fractions by column chromatography. Upon sodium dodecyl sulphate-PAGE, the enzyme showed the presence of three subunits with a molecular mass of 79,000 daltons, 49,000, and 45,000 daltons, respectively. Absorption oxidized aliphatic alcohols with a straight carbon chain except for methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidizable substrates. The apparent $K_m$ for ethanol was 1.38mM. The optimun pH and temperature were 5.0~6.0 and 32${\circ}C$, respectively. $V_2O_5$ and heavy metals such as $ZnCl_2\;and\; NiCl_2$ were inhibitory to the enzyme activity.

• Transactions of the Korean hydrogen and new energy society
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• v.19 no.6
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• pp.529-536
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• 2008

Cyanobacteria Synechocystis PCC 6803 or the extracted thylakoid membrane from this strain was appled to photosynthetic bio-electrochemical fuel cell(PBEFC) for the production of hydrogen under the illumination of 48Klux using halogen lamp. PBEFC was composed of anode, cathode and membrane between them. Electrode material was carbon paper while electron mediator and receptor were added phenazine methosulfate(PMS) and potassium ferricyanide respectively. When water and 50 mM tricine buffer and $300{\mu}M$ PMS were added to the anode under the light condition, PBEFC produced the current density $4.4{\times}10^{-5}\;mA/cm^2$, $1.4{\times}10^{-4}\;mA/cm^2$ and $2.4{\times}10^{-4}\;mA/cm^2$, respectively. And the addition of the thylakoid membrane to the system increased current density to $1.3{\times}10^{-3}\;mA/cm^2$. Two times increase of the thylakoid membrane into the anode doubled the current density to $2.6{\times}10^{-3}\;mA/cm^2$. But the current density was not increased proportionally to the amount of thylakoid membrane increased. The system was unstable to measure the electricity output due to the foam production in the anode. Addition of triton X-100 and tween 80 stabilized the system to measure the electricity output but the current density was not increased higher than $8.4{\times}10^{-4}\;mA/cm^2$ and $2.3{\times}10^{-3}\;mA/cm^2$. When the thylakoid membrane was substituted to Synechocystis PCC 6803 cells of four-day culture which has chlorophyll contents $20.5{\mu}g/m{\ell}$, maximum current density was $1.3{\times}10^{-3}\;mA/cm^2$ with $1\;k{\Omega}$ resistance.

• Journal of Environmental Science International
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• v.19 no.2
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• pp.247-254
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• 2010

Ultrasound and Surfactant aided soil washing process has been shown to be an effective method to remove diesel from soils. The use of surfactants can improve the mobility of diesel in soil-water systems by increasing solubility of adsorbed diesel into surfactant micelles. However, a large amount of surfactant is required for treatment. In addition, synthetic surfactants, specially anionic, are more toxic and the surfactant wastewater is hard to treat by conventional wastewater treatments even by AOPs. Ultrasound improves desorption of the diesel adsorbed on to soil. The mechanisms are based on physical breakage of bonds by hot spot, directly impact onto soil particle surface, the fragmentation of long-chain hydrocarbons by micro-jet and microstreaming in the soil pores. The use of ultrasound as an enhancement method in both anionic and nonionic surfactant aided soil-washing processes were studied. And all experiments were examined proceeded under CMC surfactant concentration, frequency 35 khz, power 400 W, Soil-water ratio 1:3(wt%), particle size 0.24 ~ 2mm and initial diesel concentration. 20,000 mg/kg. Combination with ultrasound showed significant enhancements on all the processes. Especially, nonionic surfactant Triton-X100 with ultrasound showed remarkable enhancements and diesel removal rate enhanced by ultrasound helps desorpting of surfactant adsorbed onto soils which prevented decreasing surfactant activity.

• Journal of the Korean Chemical Society
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• v.35 no.2
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• pp.151-157
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• 1991

A sensitive stripping voltammetric study of the complex of indium with 8-hydroxyquinoline at a hanging mercury drop electrode was investigated in 0.1M acetate buffer solution. The effects of various analytical conditions on the reduction peak current of the adsorbed complex were discussed. Optimal analytical conditions were found to be the ligand concentration of $2 {\times}10^{-5}$M, solution pH 4.75, scan rate of 10 mV/s, deposition potential of -0.450V, a deposition time of 90 second. Interferences by other trace metals and Triton X-100 were also discussed. Detection limit was 0.2 ppb of indium after 90 sec. Deposition time, and the relative standard deviation(n = 10) at 4 ppb was 3.2%.

• Journal of Environmental Health Sciences
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• v.24 no.2
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• pp.74-79
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• 1998

This study was conducted to evaluate the surfactant solutions which influence solvent extraction from light hydrocarbon contaminated soils. Nine characteristic compounds were studied: benzene, toluene, ethylbenzene, o-xylene, mxylene, p-xylene, n-propylbenzene, 1,2,4-trimethylbenzene, and n-butylbenzene which were found in gasoline. Adsee-799 and Witbreak DRA-22 showed some extractive capacity for light hydrocarbons from soil. There was no added advantage obtained by using other surfactants in this study. No removal of contaminants from soil was observed when the surfactant concentration was 0.5 percent or below. When the surfactant concentration was 4 percent, the average recovery for some hydrocarbons was 10.8 percent, which was the best obtained at these levels. There was 10 percent surfactant contribution for methanol extraction from soil with the Witbreak DPG-482 and Witbreak DRA-22. This study provided a useful screening technique for procedures that can be used to remediate soils contaminated with light hydrocarbons.

• KSBB Journal
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• v.17 no.2
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• pp.176-181
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• 2002

Astaxanthin (3,3＇-dihydroxy-${\beta}$, ${\beta}$-carotene-4-4＇-dione), a natural pigment of pink to red color, is widely distributed in nature particularly in the skin layer of salmonoids and the crust of shrimp, lobster, etc. Recently, it was produced from the yeast culture of Phaffia rhodozyma. Because of its high thermal stability and antioxidant functionality, its applications can be extended into food, cosmetics, and pharmaceutical ingredient beyond the traditional feed additive. Because of its very high lipophilicity, astaxanthin has been extracted traditionally by strong organic solvents such as chloroform, petroleum ether, acetone, etc. In this study, we developed a surfactant-based solubillization system for astaxanthin, and used it to extract astaxanthin from disrupted yeast cells. Among Tween 20, Triton X-100 and SDS, Tween 20 was identified as the most suitable surfactant in terms of extraction capacity and safety. The ethylene oxide group of Tween 20 was identified as the most significant factor to increase the HLB value that determined the extraction capacity. The effects of micelle formation condition, such as the molar ratio of astaxanthin and Tween 20, pH, and ionic strength were also investigated. pH and ionic strength showed no significant effects. The optimal molar ratio between astaxanthin and Tween 20 was 1 : 12. Antioxidant activity of astaxanthin was higher than ${\beta}$-carotene and ${\alpha}$-tocopherol. Astaxanthin in the crude extract from the yeast cell was more resistant to air and/or light degradation than pure astaxanthin, probably because of the presence of other carotenoids and lipids.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.604-610
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• 2007

A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 bp coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at $30^{\circ}C$, and was unstable at temperatures higher than $30^{\circ}C$, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24h incubation at $4^{\circ}C$. The addition of $Ca^{2+}\;and\;Mg^{2+}$ enhanced the enzyme activity of LipA1, whereas the $Cd^{2+},\;Zn^{2+},\;CO^{2+},\;Fe^{3+},\;Hg^{2+},\;Fe^{2+},\;Rb^{2+}$, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate $(C_{14}\;acyl\; groups)$.

• The Journal of Internal Korean Medicine
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• v.34 no.3
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• pp.278-288
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• 2013

Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.