• Title/Summary/Keyword: Isoelectric point

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Purification and characterization of a xylanase from alkalophilic cephalosporium sp. RYM-202

  • Kyu, Kang-Myoung;Kwon, Tae-Ik;Rhee, Yuung-Ha;Rhee, Young-Ha
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.109-114
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    • 1995
  • Alkalophilic Cephalosporium sp. RYM-202 produced multiple xylanases extracellularly. One of these xylanases was purified to electrophoretical homogeneity by chromatography with DEAE-Sephadex A-50, Sephacryl S-200 HR and Superose 12 HR. The purified xylanase differed from most other microbial xylanases in that it had low-molecular weight and acidic isoelectric point. The molecular weight of the xylanase in that it had low-molecular weight and acidic isoelectric point. The molecular weight of the xylanase was 23 kDa by SDS-polyacrylamide electrophoresis and 24 kDa by gel permeation chromatography, and the isoelectric point was 4.3. The xylanase had the highest activity permentation chromatography, and the isoelectric point was 4.3. The xylanase had the highest activity permeation chromatography, and the isoelectric point was 4.3. The xylanase had the highest activity at pH 8.0 and 50 .deg.C. It was stable over a wide range of pH and retained more than 80% of its original activity after 24 h of incubation even at pH 12. The Km values of this enzyme on birchwood xylan and oat spelts xylan were 2.33 and 3.45 mg/ml, respectively. The complete inhibition of the enzyme of n-bromosuccinimide suggests the involvement of tryptophan in the active site. The sylanase lacked activity towards crystalline cellulose and carboxymethyl cellulose.

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Interfacial Characterization of $\beta$-Sialon Powder Prepared from Hadong Kaolin (하동 카올린으로부터 제조한 $\beta$-Sialon 분체의 계면특성)

  • 임헌진;이홍림
    • Journal of the Korean Ceramic Society
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    • v.29 no.7
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    • pp.551-557
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    • 1992
  • X-ray diffraction patterns, IR spectra and zeta-potentials of silicon nitride and $\beta$-Sialon powders were investigated before and after surface manipulations. $\beta$-Sialon powder was produced from Hadong Kaolin by the carbothermic reduction and simulataneous nitridation. Isoelectric points of as-prepared Si3N4 and $\beta$-Sialon powders were 8.4 and 7.4, respectively. After both silicon nitride and $\beta$-Sialon powders were oxidized at 80$0^{\circ}C$ for 24 h in air, the isoelectric points of these powders corresponded to that of silica (pH=3). I case of the addition of Darvan C as deflocculant, its isoelectric point was 3 and zeta-potential was nearly constant in the range of pH 5~12. When SN 7347 was used as deflocculant, its isoelectric point was 8.3 and zeta-potential over -156 mV was measured above pH 11.

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Characteristics of the Turbidity Change of Clay Particulate Matter according to Its Surface Electrokinetic Behavior (점토성 광물입자의 표면 전기적 거동에 따른 탁도 변화 특성)

  • O, Sejin;Kim, Dong-Su
    • Journal of Korean Society on Water Environment
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    • v.26 no.2
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    • pp.326-331
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    • 2010
  • Montmorillonite is one of representative inorganic clay particles. As the characteristics of clay particulate matter in aqueous environment determine the efficiencies of wastewater treatment and some industrial operations, it is essential to understand its aquatic behavior in relation with turbidity. The change of electrokinetic potential of montmorillonite suspension shows that it tends to negatively increase as the pH of suspension increases. In addition, it is observed that its potential is around 0mV when the solution pH is ca. 5. The turbidity of suspension is shown to be very low when pH is lower than its isoelectric point. However, the turbidity gradually enhances according to beyond isoelectric point. These results reveal that the correlation between electrokinetic potential and turbidity for clay mineral suspension is peculiar which should be fundamentally considered for systematic treatment of wastewater.

Retrospective analyses of the bottleneck in purification of eukaryotic proteins from Escherichia coli as affected by molecular weight, cysteine content and isoelectric point

  • Jeon, Won-Bae
    • BMB Reports
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    • v.43 no.5
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    • pp.319-324
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    • 2010
  • Experimental bioinformatics data obtained from an E. coli cell-based eukaryotic protein purification experiment were analyzed in order to identify any bottleneck as well as the factors affecting the target purification. All targets were expressed as His-tagged maltose-binding protein (MBP) fusion constructs and were initially purified by immobilized metal affinity chromatography (IMAC). The targets were subsequently separated from the His-tagged MBP through TEV protease cleavage followed by a second IMAC isolation. Of the 743 total purification trials, 342 yielded more than 3 mg of target proteins for structural studies. The major reason for failure of target purification was poor TEV proteolysis. The overall success rate for target purification decreased linearly as cysteine content or isoelectric point (pI) of the target increased. This pattern of pI versus overall success rate strongly suggests that pI should be incorporated into target scoring criteria with a threshold value.

Surface Potential Behavior of Nano $CeO_2$ Particles in Aqueous Media (수계분산매체에서 나노 $CeO_2$ 입자의 계면전위 거동)

  • 이태원;백운규;최성철;이상훈;임형섭;김철진
    • Journal of the Korean Ceramic Society
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    • v.37 no.7
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    • pp.721-725
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    • 2000
  • In this study, the dispersion stability of nano-sized CeO2 particles, synthesized by hydrothermal method in aqueous was evaluated from observing the surface potential behavior of CeO2 particle synthesized by solid state reaction. The isoelectric point(IEP) of nano-sized CeO2 synthesized by hydrothermal synthesis was found to be pH 9 contrary to the isoelectric point of micro-sized CeO2 synthesized by solid state reaction at pH 6.7. IEP was shifted to pH 2.0 as the addition of D-3019 from 0.1 to 1.0 wt%. The surface potential of CeO2 particles synthesized by hydrothermal synthesis was reduced as the addition of B-1001 used as a binder without change of IEP because the absorption of B-1001 polymer on the CeO2 particles shifted the shear plane of CeO2 particles outward away from the surface. This surface potential behavior was well correlated with the dispersion stability of slurry.

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Measurement of Isoelectric Point of Amine Oxide Zwitterionic Surfactant by QCM (Quartz Crystal Microbalance) (QCM (Quartz Crystal Microbalance)을 활용한 Amine Oxide 양쪽성 계면활성제의 등전점 측정에 관한 연구)

  • Kim, JiSung;Park, JunSeok;Lim, JongChoo
    • Applied Chemistry for Engineering
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    • v.20 no.1
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    • pp.9-14
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    • 2009
  • A zwitterionic surfactant shows not only detergency but also softening effect since it shows characteristics of a nonionic or an anionic surfactant above an isoelectric point, while showing characteristics of a cationic surfactant below an isoelectric point. Therefore, a zwitterionic surfactant can serve as a dual function surfactant by a single molecule through the interconversion of cleaning and softening effects depending on pH of the aqueous solution. In this study, the dual function characteristics of an amine oxide zwitterionic surfactant were investigated by measuring the zeta potential and the isoelectric point using quartz crystal microbalance (QCM). In addition, the physical properties of an amine oxide surfactant such as critical micelle concentration, surface tension, interfacial tension, contact angle and viscosity were measured and phase behavior study was also performed. The isoelectric point of an amine oxide surfactant determined by zeta potential measurement was near 7.35 and that obtained by QCM experiment was about 7.4, where both results were found to be close to the value reported in the literature.

Measurement of Isoelectric Point of Betaine Zwitterionic Surfactant by QCM(Quartz Crystal Microbalance) (QCM(Quartz Crystal Microbalance)을 이용한 Betaine 양쪽성 계면활성제의 등전점 측정)

  • Kim, Ji Sung;Park, Jun Seok;Lim, Jong Choo
    • Korean Chemical Engineering Research
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    • v.47 no.1
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    • pp.31-37
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    • 2009
  • A zwitterionic surfactant shows not only detergency but also mildness effect since it shows characteristics of a nonionic or an anionic surfactant above an isoelectric point, while showing characteristics of a cationic surfactant below an isoelectric point. Therefore, a zwitterionic surfactant can serve as a dual function surfactant by a single molecule through the interconversion of cleaning and softening effects depending on pH variations of the aqueous solution. In this study, physical properties of betaine surfactant such as CMC, surface tension, interfacial tension, contact angle and viscosity were measured and phase behavior study was performed. Also dual function characteristics of betaine zwitterionic surfactant were investigated by measuring an isoelectric point using QCM(quartz crystal microbalance) and zeta potential measurement. The CMC of betaine surfactant was near $10^{-4}mol/L$ and the surface tension at CMC was about 32 mN/m. The interfacial tension between 1 wt% aqueous solution and n-decane measured by spinning drop tensiometer at pH 2~10 resulted in an increase in interfacial tension until pH 5 and a decrease with pH after 5 and equilibration time showed the similar trend with an increase in pH. The isoelectric point of betaine surfactant measured by QCM experiment was found to exist between 3.0 and 3.3, which is the same as the result determined by zeta potential measurement.

Measurement of an Isoelectric Point and Softness of a Zwitterionic Surfactant (양쪽성 계면활성제의 등전점 및 유연력 측정에 관한 연구)

  • Lim, Jongchoo;Kim, Jisung;Mo, Dahee;Lee, Jinsun
    • Applied Chemistry for Engineering
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    • v.23 no.1
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    • pp.112-118
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    • 2012
  • In this study, physical properties of synthesized DE7-OSA82-AO and DEP52-OSA82-AOQ82 zwitterionic surfactants were measured such as critical micelle concentration, surface tension, interfacial tension, contact angle and viscosity. Phase behavior study was also performed. The dual function characteristics of a zwitterionic surfactant were investigated by determining an isoelectric point, which was attained using zeta potential measurements and QCM (quartz crystal microbalance) experiments. The isoelectric point of DE7-OSA82-AO surfactant determined by the zeta potential measurement and QCM experiment was about 7.2 and 7.4, respectively. On the other hand, the isoelectric point of DEP52-OSA82-AOQ82 surfactant determined by the zeta potential measurement and QCM experiment was about 10.9 and 11.0, respectively. The frictional property measured using an automated mildness tester showed that DE7-OSA82-AO surfactant can provide a good softening effect at an acidic or neutral condition. On the other hand, DEP52-OSA82-AOQ82 was found to provide a good softening effect to a fabric surface at a pH below its isoelectric point of 11.

Purification of Bovine Pregnancy-Associated Proteins by Two-Dimensional Gel Electrophoresis

  • Hwang, Sun-Il;Lim, Jin-Kyu
    • BMB Reports
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    • v.32 no.5
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    • pp.445-450
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    • 1999
  • We purified and characterized a bovine pregnancy-associated protein in pregnant cow urine using two-dimensional gel electrophoresis. Urine from cows was collected according to their status of pregnancy and non-pregnancy. Proteins in the cow urine were fractionated with 50% ammonium sulfate prior to two-dimensional gel electrophoresis. Proteins separated on the gels were compared in terms of expression level and new expression by molecular mass and isoelectric point. We localized two pregnancy-associated protein spots on the gels at molecular masses of 24 kDa and 20 kDa and isoelectric points of 5.5 and 5.7, respectively. Likewise, two non-pregnancy specific proteins were localized at 27 kDa and 28 kDa with isoelectric points of 5.7 and 5.9, respectively. To rule out the possibility that environmental or genetic factors might influence the expression of the proteins, we demonstrated the pregnancy-associated expression of the proteins in two-dimensional gels with pregnant urine taken from cows raised in a different institute. The pregnancy-associated protein with molecular mass of 20 kDa and isoelectric point of 5.7, namely spot 2, was microsequenced and found to be highly homologous to the bovine collagen alpha 1 chain.

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Electrochemical modification of the porosity and zeta potential of montmorillonitic soft rock

  • Wang, Dong;Kang, Tianhe;Han, Wenmei;Liu, Zhiping;Chai, Zhaoyun
    • Geomechanics and Engineering
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    • v.2 no.3
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    • pp.191-202
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    • 2010
  • The porosity (including the specific surface area and pore volume-diameter distribution) of montmorillonitic soft rock (MSR) was studied experimentally with an electrochemical treatment, based on which the change in porosity was further analyzed from the perspective of its electrokinetic potential (${\zeta}$ potential) and the isoelectric point of the electric double layer on the surface of the soft rock particles. The variation between the ${\zeta}$ potential and porosity was summarized, and used to demonstrate that the properties of softening, degradation in water, swelling, and disintegration of MSR can be modified by electrochemical treatment. The following conclusions were drawn. The specific surface area and total pore volume decreased, whereas the average pore diameter increased after electrochemical modification. The reduction in the specific surface area indicates a reduction in the dispersibility and swelling-shrinking of the clay minerals. After modification, the ${\zeta}$ potential of the soft rock was positive in the anodic zone, there was no isoelectric point, and the rock had lost its properties of softening, degradation in water, swelling, and disintegration. The ${\zeta}$ potential increased in the intermediate and cathodic zones, the isoelectric point was reduced or unchanged, and the rock properties are reduced. When the ${\zeta}$ potential is increased, the specific surface area and the total pore volume were reduced according to the negative exponent law, and the average pore diameter increased according to the exponent law.