• Journal of the Optical Society of Korea
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• v.20 no.3
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• pp.431-434
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• 2016

Tris-HCl buffer solution is extensively used in biochemistry and molecular biology to maintain a stable pH for biomolecules such as nucleic acids and proteins. Here we report on the high-precision THz dielectric spectroscopy of a 10 mM Tris-HCl buffer. Using a double Debye model, including conductivity of ionic species, we measured the complex dielectric functions of Tris-HCl buffer. The fast relaxation time of water molecules in Tris-HCl buffer is ~20% longer than that in pure water while the slow relaxation time changes little. This means that the reorientation dynamics of Tris-HCl buffer with such a low Tris concentration is quite different from that of pure water.

• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.613-621
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• 2018

In this study, interface-assembled carbonyl reductase (IACR) was prepared and used in the synthesis of S-licarbazepine in a toluene/Tris-HCl biphasic system. The carbonyl reductase (CR) was conjugated with polystyrene to form a surfactant-like structure at the interface of the toluene/Tris-HCl biphasic system. The interface-assembled efficiency of IACR reached 83% when the CR (180 U/mg) and polystyrene concentration were $8{\times}10^2g/ml$ and $3.75{\times}10^3g/ml$, respectively. The conversion reached 95.6% and the enantiometric excess of S-licarbazepine was 98.6% when $3.97{\times}10^6nmol/l$ oxcarbazepine was converted by IACR using 6% ethanol as a co-substrate in toluene/Tris-HCl (12.5:10) at $30^{\circ}C$ and $43{\times}g$ for 6 h. IACR could be reused efficiently five times.

• Korean Journal of Veterinary Research
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• v.33 no.4
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• pp.623-632
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• 1993

This study was conducted to develop a sensitized latex-antigen for serodiagnosis of toxoplasmosis in animals. Tachyzoites of T gondii(RH-strain) harvested from mouse peritoneal cavity were purified through the filtraton of polycarbonate membrane(pore size, $3.0{{\mu}m}$, Costar Co.) and disrupted by ultrasonicator. The tachyzoite suspension was ultracentrifuged for 30 min at $60,000{\times}g(4{^{\circ}C})$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $0.8{{\mu}m}$ in diameter(Sigma) were used for the preparation of sensitized latex-antigen suspension. The several parameters including the preparation conditions, incubation buffer. serum dilution buffer and stability of agglutination reactions were evaluated and the results obtained were summarized as follows : 1. The antigen consisting of a water-lysate of T gondii tachyzoites was adsorbed onto polystyrene latex particles of $0.8{{\mu}m}$ in diameter by adding a latex suspension to an equal volume of diluted antigen solution and by incubating the mixture at $37{^{\circ}C}$ under different conditions. 2. The optimum incubation buffer used for the antigen sensitization was 0.1M Tris-HCl buffer(pH 8.0). 3. The optimum serum dilution buffer used for the latex agglutination test was 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300 mM NaCl. But 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300-600 mM NaCl, 0.5% BSA and 0.01% Tween-20 improved the agglutination pattems and cleared the background of microplate well without the effects on L.A titer. 4. The time required for antigen sensitization was 40 and 60 min in incubation buffer(pH 8.0) at $37{^{\circ}C}$. But the optimun time for antigen sensitization was min at $37{^{\circ}C}$. 5. The optimun quantity of antigen absorbed on latex particles for proper agglutination was the range of 20 to $32{\mu}g$ of latex particles. 6. The optimun concentration of the latex-antigen suspension for the proper agglutination reaction was determined as 0.2%(w/v). 7. The specificity, rapidity and simplicity of the latex-particle agglutination test suggested that it might be adaptable to large scale serum screening.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.357-365
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• 1989

Induction conditions for autolysis and autoplast formation of thermophilic Clostridium thermohydrosulfuricum were studied. The cells in the initial exponential growth phase were well autolysed in Tris-HCl buffer or inorganic buffers containing univalents, such as $K^{+}$ and $Na^{+}$ , and chemicals such as cysteine-HCl, sorbitol and glycerol. Meanwhile, autolysis induction was slightly inhibited by divalents, such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$, and strongly by divalents, such as $Fe^{2+}, Cu^{2+}$ and citric acid. The autolysis was stimulated when the cells were grown in the medium containing ampicillin that inhibites cell wall synthesis, meanwhile, it was slightly inhibited by nucleic acids and protein synthesis inhibitors. The optimal pH and temperature for the induction of autolysis were 7.5 and $60^{\circ}C$, respectively. On the other hand, the cells were autoplasted without lysozyme treatment during autolysis due to the stabilization of protoplasmic membrane in the presence of divalents such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$. Autoplast formation was mostly induced at $37^{\circ}C$ in 50mM Tris-HCl buffer (pH 7.5) containing 20 mM $MgCl^{2}$ and 0.3 M glycerol, and in the late exponential growth phase growing cell.

• Membrane Journal
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• v.18 no.4
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• pp.306-316
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• 2008

The adsorption characteristics of L-tryptophan of PES-BSA affinity membranes prepared by using electro-spinning were investigated. It was found that the fiber diameters could be controlled by the comparisons of fiber diameters prepared by various spinning conditions through FESEM and Image Analyzer. Darcy's permeability constants, mechanical properties and hydrophobic properties were enhanced since micro-fibers increased by increasing the composition ratio of HFB and BSA. The elution capacity of L-tryptophan in borate-DMSO buffer solution was higher than that in tris-HCl buffer solution, while the elution capacity of A 7906 type BSA was higher than A 8022 type BSA. This is due to the characteristics of BSA type, i. e. higher purity and uniform molecular weight and better pH stability of A 7906 type BSA than those of A 8022 type BSA.

• Journal of the Korean Chemical Society
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• v.36 no.3
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• pp.393-404
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• 1992

The response properties of continuous automated system using an enzyme reactor for determination of urea were investigated. The enzyme reactor was constructed to packed-bed form which filled with nylon-6 beads (42∼48 mesh), which immobilized urease with glutaraldehyde, in teflon tube (2 mm I.D., 20 cm length). The system was composed of the enzyme reactor, gas dialyzer, and tublar PVC-nonactin membrane ammonium ion-selective electrode as an indicator electrode in serial order. The response characteristics of this system were as follows. That is, the concentration range of linear response, slope of linear response, detection limit, and conversion percentage were $5.5{\times}10^{-6}$∼$2.4{\times}10^{-3}M$, 57.8 mV/decade, $1.5{\times}10^{-6}$, and 80.8%, respectively. The optimum buffer and life time of urease reactor were 0.01M Tris-HCl buffer solution (pH 7.0∼7.8) and 0.01M phosphate buffer solution (pH 6.9∼7.5) and about 150 days, respectively. And the urease reactor had no interferences of the other physiological materials.

• KSBB Journal
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• v.15 no.6
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• pp.605-608
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• 2000

Oligodeoxynucleotides (ODNs) were separated by high-performance membrane chromatography (HPMC), a combined system of chromatography and membrane. The separation mechanism involved anion-exchange, and the stationary phase was cation CIM (Convective Interaction Media) DEAE disk (16${\times}$3 mm). Two types of mobile phase were used, buffer A (20mM Tris-HCl, pH 7.4) and buffer B (buffer A + 1M NaCl). As the amount of NaCl dissolved in buffer linearly increased, the retention time shortened, which enabled a gradient elution mode. Based on the number of theoretical plates and resolution observed, the optimum mobile phase and operating condition (Buffer A/Buffer B=50/50 - 20/80 vol%, gradient time 2 min) were experimentally determined. In this experimental condition, ODNs were separated within 2 min at a mobile phase flow rate of 6 ml/min.

• Korean Journal of Microbiology
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• v.23 no.3
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• pp.177-183
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• 1985

Some properties of an extracellular cytosine deaminase produced from Arthrobacter sp.JH-13 were examined after 20-80% of ammonium sulfate fractionation. Among some substrates, this enzyme utilized cytosine and 5-fluorocytosine as a substrate. The optimum pH and temperature for the activity of this enzyme were found to be near 8.0 and $40^{\circ}C$, respectively. The ensyme was more stable in 0.2M of Tris-HCl buffer than 0.2M of potassium phosphate buffer. The enzyme was generally stable below $50^{\circ}C$, but inactivated completely at $70^{\circ}C$. 1mM of $Fe^{3+},\;K^+\;and\;Na^+$ increased the enzyme activity, but 0.01mM of $Co^{2+},\;Cu^{2+},\;Ni^{2+},\;Hg^{2+},\;Ag^{2+},\;Zn^{2+},\;Ba^{2+},\;and\;Mg^{2+}$ markedly inactivated the enzyme activity. 0.1mM of p-chloromercuribenzoate, trichloroacetic acid, and N-ethylmaleimide compleyely inhibited the enzyme activity, but 0.1mM of 2-mercaptoethanol slightly increased the enzyme activity.

• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.495-500
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• 1986

Enzymatic micro-assay methods were studied those were capable of determining ammonia down to 10$^{-5}$M(0.01 $\mu$mole/ml) in the presence of other nitrogenous compounds such as protein and amino acid. Microquantities of ammonia (0.01-0.1 $\mu$mole) could be determined indirectly by measuring phosphorous, one of the products of the enzymatic reaction catalyzed by glutamine synthetase. In this reaction, L-glutamate, ATP and ammonium chloride were used as substrates, and phosphorous was formed in propotion to the concentration of ammonium chloride In the reaction mixture. Another procedure was examined in which glutamine synthetase reaction coupled with pyruvate kinase and lactate dehydrogenase reactions was used. One mililiter of the assay mixture contained; phosphoenol pyruvate, 3 mM, L-glutamate, 10 mM; ATP, 1mM: MgSO$_4$, 20 mM: KCl, 75mM: NADH, 0.2mM: Tris-HCl buffer(pH 7.0), 100mM; pyruvate kinase, 10 U: lactate dehydrogenase, 12 U and glutamine synthetase, 4 U. After preincubation for 20 min at 3$0^{\circ}C$, NH$_4$Cl was added and the rates of NADH oxidation were followed at 340nm. The effective range of this method was proved to be from 0.01 to 0.05 $\mu$mole/$m{\ell}$. Glutamine synthetase reaction coupled with glutamate synthase reaction could also be effectively used for determining microquantities of ammonia. The one mililiter assay mixture contained; ATP, 5mM: L-glutamate, 5mM; L-ketoglutarate, 5mM; MgCl$_2$, 15mM; NADPH, 0.15mM; Tris-HCl buffer(pH 7.0); 100mM; glutamine synthetase, 1U and glutamate synthase, 0.5U. After preincubation for 20min at 3$0^{\circ}C$ NH$_4$Cl was added and the rates of NADPH oxidation were followed at 340nm. The effective range of this procedure was appeared to be from 0.01 to 0.05$\mu$mole/$m{\ell}$.

• Journal of Plant Biology
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• v.29 no.2
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• pp.85-94
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• 1986

Canavanine content of the cotyledons of Canavalia lineata decreased gradually during germination and growth of seedlings but continued to increase in roots and leaves. After abscission of cotyledons, canavanine content of leaves depleted competely. The activity of canavanase could be detected in leaves and roots, but not in cotyledons. High arginase activity was observed in the cotyledons of seeds at the earlyimbibition period. During the growth of seedlings, cotyledonary canavanine appeared to be transported to the growing of seedlings where it could be utilized through nitrogen metabolic pathways. In crude cell-free extracts of leaves, maximum activities of canavanase or arginase appeared in 30mM Tris-HCl buffer (pH 9.0) or 30mM NaHCO3 buffer (pH 10.0), respectively. The activities of these two enzymes differed from each other when treated with Co2+ or Mn2+. These results support the idea that canavanase and arginase might be different enzymes.