• Title/Summary/Keyword: Triton X

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Enhanced pectinase and β-glucosidase enzyme production by a Bacillus subtilis strain under blue light-emitting diodes

  • Elumalai, Punniyakotti;Lim, Jeong-Muk;Oh, Byung-Teak
    • 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.

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Purification and Properties of a Membrane-bound Alcohol Dehydrogenase from Acetobacter sp. HA (Acetobacter sp. HA로부터 Membrane-bound Alcohol Dehydrogenase의 정제 및 특성)

  • Yoo, Jin-Cheol;Sim, Jung-Bo;Kim, Heung-Keun;Chun, Hong-Sung;Kim, Sung-Jin
    • 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.

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Electricity Generation Using Cyanobacteria Synechocystis PCC 6803 in Photosynthetic Bio-Electrochemical Fuel Cell (남조류 Synechocystis PCC 6803을 이용한 생물전기화학적 물분해 전기 생산)

  • Kim, Min-Jin;Oh, You-Kwan;Kim, Mi-Sun
    • 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.

The Effect of Ultrasound Application to Anionic/Non-ionic Surfactant Aided Soil-washing Process for Enhancing Diesel Contaminated Soils Remediation (디젤오염토양 복원 효율 증진을 위한 음이온/비이온 계면활성제 토양세척공정에 초음파 적용 영향)

  • Cho, Sang-Hyun;Son, Young-Gyu;Nam, Sang-Geon;Cui, Ming-Can;Khim, Jee-Hyeong
    • 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.

Magnetic Orientations of Bull Sperm Separated into Head and Flagellum Treated by DTT or Heparin

  • Suga, D.;Shinjo, A.;Kurnianto, E.;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.2
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    • pp.167-175
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    • 2000
  • This paper describes the magnetic orientation of bull sperm separated into the head and the flagellum treated by DTT or heparin in a 5,400G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4^{\circ}C$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the head area at 1, 3 and 6 days. After measuring the area, each sample was exposed to a 5,400G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the sperms were separated into the head and the flagellum through the DTT treatment. Almost of the separated heads showed that their long axis oriented perpendicularly to the magnetic lines of force, and most of the long axis perpendicularly oriented heads showed that their flat plane oriented perpendicularly in a 5,400G magnetic field. Also, the demembranation of the head tended to increase those perpendicular orientations, while those perpendicular orientations of the head declined with the decondensation of the sperm nuclei. These findings suggest that strong magnetic anisotropy for the perpendicular orientation of the long axis and the flat plane of the head occurs in the sperm nuclei in a 5,400G magnetic field. The separated flagellum showed lower parallel orientation, and the separated and demembranated flagellum showed parallel orientation to the magnetic lines of force in this magnetic field. These findings suggest that weak magnetic anisotropy of the parallel orientation of the flagellum occurs in the inside components in a 5,400G field.

Adsorptive Stripping Voltammetry of the Indium-8-Hydroxyquinoline Complex (Indium-8-Hydroxyquinoline 착물에 관한 흡착벗김전압전류법적 연구)

  • Se Chul Sohn;Tae Yoon Eom;Moo Yul Suh;Ki-Suk Jung
    • 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%.

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Effect of Surfactant on Solvent Extraction for Light Hydrocarbon from Soils (토양중 Light Hydrocarbon의 용매추출에 미치는 계면활성제의 영향)

  • Hwang, Keon-Joong;Atalay, Asmare
    • 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.

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Solubillzation and Extraction Of Antioxidant Astaxanthin by Micelle Formation from Phaffia rhodozyma Cell Homogenate (Phaffia rhodozyma 세포파쇄액으로부터 항산화제 Astaxanthin의 미셀 형성을 통한 가용화 및 추출)

  • Kim, Young-Beom;Ryu, Kang;Lim, Gio-Bin;Lee, Eun-Kyu
    • 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.

Isolation of an Acinetobacter junii SY-01 Strain Producing an Extracellular Lipase Enantioselectively Hydrolyzing Itraconazole Precursor, and Some Properties of the Lipase

  • Yoon, Moon-Young;Shin, Pyong-Kyun;Han, Ye-Sun;Lee, So-Ha;Park, Jung-Keug;Cheong, Chan-Seong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.97-104
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    • 2004
  • Water-sludge bacteria were screened to find a lipase enantioselectively hydrolyzing itraconazole precursor, which is well known as the starting material of antifungal drug agents. A bacterial strain was isolated and identified as Acinetobacter junii SY-01. After the strain was cultivated, the enzyme was purified 39.4-fold using ultrafiltration and gel filtration through a Sephadex G-100 chromatographic column and the activity yield was 34.9%. The molecular weight of the enzyme was about 40 kDa, as measured by SDS-PAGE, and the optimum pH was 7.0- 9.0 and stable at pH 6.0- 9.0. The optimum temperature was 45- $5^{\circ}C$, and 73% of the enzymes activity remained after incubation at 70% for 1 h. Enzyme activity was enhanced by gall powder, sodium deoxycholate, a cationic detergent Tween 80, and a non-ionic detergent Triton X-100, but was markedly inhibited by metal ions such as $Hg^{2+},Cu^{2+},Ni^{2+}/,Ca^{2+}$, and an anionic-surfactant sodium dodecylsulfate. The $K_{m}$ values for (R)- and (S)-enantiomers of the itraconazole precursor were 0.385 and 21.83 mM, respectively, and the $V_{max} values ($\mu$Mㆍmin^{-1}.)$ were 6.73 and 6.49, respectively. The acetyl group among the different acyl moieties of itraconazole precursor showed the highest enantioselectivity for the hydrolysis by the Acinetobacter junii SY-01 lipase, and the lipase from Acinetobacter junii SY-01 displayed better enantioselectivity than that of commercially available lipases and esterases.

Cloning, Expression, and Characterization of a Cold-Adapted Lipase Gene from an Antarctic Deep-Sea Psychrotrophic Bacterium, Psychrobacter sp. 7195

  • Zhang, Jinwei;Lin, Shu;Zeng, Runying
    • 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 $(C_{14}\;acyl\; groups)$.