The peptidyl prolyl sis-trans isomerase (PPIase, EC 5.2.1.8) from bacillus stearothermophilus was extracted from the cells treated with by lysozyme. PPIase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPIase was estimated as 18kDa by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0∼8.0. The enzyme was considerably stable after heat treatment at 60$\^{C}$ for 30minutes, and the enzyme was quite stable up to 65$\^{C}$. The presence of the PPIase in the refolding solution accelerated the isomerization rate of the assay peptide. PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-l primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPI-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus.
The optimum conditions for measuring cadmium content of less than 0.2ppm by flame atomic absorption spectrophotometry were investigated. The cadmium in urine was extracted by APDC-MIBK for the analysis by atomic absorption spectrophotometry after ashing them by a wet method. 1. Optimum conditions by APDC-MIBK and DDTC-MIBK extractions. The acidic aqueous solution was prepared with appropriate amount of 0.IN nitric acid, 5ml of 25% (W/V) sodium potasstum tartarate, 10ml of saturated ammonium sulfate, and 2ml of 2% APDC(or 1 ml of 5% DDTC) chelating agent. The total volume of solution was adjusted to 55 ml and pH to $2{\sim}10$ (or$7{\sim}10$). The aqueous solution was extracted with 10ml MIBK. Concentration of Triton X-100 did not effect the absorbance for APDC-MIBK extraction of cadmium, but absorbance decreased as the concentration increased for DDTC-MIBK extraction. The sensitivity and detection limits for the cadmium determination from APDC-MIBK extraction were 0.0038ppm and 0.0102, 0.0022ppm and 0.0116 for DDTC-MIBK, and 0.0132ppm and 0.0034 for 0.1N nitric acid. APDC-MIBK and DDTC-MIBK extractions were 3 times higher than 0.1N nitric acid for the sensitivity. 2. Excretion of cadmium in 24-hour urine by APDC-MIBK extraction. Determination of cadmium in urine by atomic absorption spectrophotometry of A.A. (Cd=2 mA) mode and B.C. (Cd=4 mA) mode and B.C. (Cd=4mA, $D_2=20mA$) mode showed some difference (p<0.05). The difference of cadmium determination and recovery according to method of standard additions and standard calibration curve method in urine was not significant (p>0.05, $93.48{\pm}11.78%,\;94.83{\pm}22.00%$). Excretion of cadmium in 24-hour urine collection from normal person and variance analysis within measurement variation was not significant (p>0.05), but between interindividual was significant (0.05). Determination of cadmium content by two different methods of flame atomic absorption spectrophotometry and dithizone colorimetry showed that the results from the two methods can be described by a regression line with a good correlation (y=1.0153x-0.2927, x=Cd by D.C., y=Cd by A.A.S., $r=0.8651^*$, p<0.01).
Swain Basudev;Jeong Jinki;Kim Min Seuk;Lee Jae-chun;Sohn Jeong-Soo
Resources Recycling
/
v.14
no.6
s.68
/
pp.28-36
/
2005
A hydrometallurgical process to leach cobalt from the waste cathodic active material, $LiCoO_{2}$, and subsequently to separate it by solvent extraction was developed. The optimum leaching conditions for high recovery of colbalt and lithium were obtained: 2.0 M sulfuric acid, 5 $vol.\%$ hydrogen peroxide, $75^{\circ}C$ leaching temperature, 30 minutes leaching time and an initial pulp density of 100 g/L. The respective leaching efficiencies for Co and Li were $93\%$ and $94.5\%$. About $85\%$ Co was extracted from the sulfuric acid leach liquor containing 44.72 g/L Co and 5.43 g/L Li, using 1.5 M Cyanex272 as an extractant at the initial pH 5.0 and in organic to aqueous phase ratio of 1.6:1 under the single stage extraction conditions. The Co in the raraffinate was completely extracted by 0.5 M Na-Cyanex272 at the inital pH 5.0, and an organic to aqueous phase ratio of 1;1. The cobalt sulfate solution of higher than $99.99\%$ purity could be recovered from waste $LiCoO_{2}$, using a series of hydrometallurgical processes: sulfuric acid leaching of waste $LiCoO_{2}$- solvent extraction of Co by Na-Cyanex 271 - scrubbing of Li by sodium carbonate solution - stripping of Co by sulfuric acid solution.
It has been demonstrated that intracoronal bleaching of pulpless teeth may result in cervical root resorption. Several authors postulated that bleaching agents such as hydrogen peroxide penetrated through the dentinal tubules to damage the surrounding tissues that cause cervical root resorption. The purpose of this study was to suggest on in vitro model for direct determination of hydrogen peroxide penetration through CEJ during nonvital bleaching. In addition, this model permit the quantification of the amount of hydrogen peroxide penetrated during the procedure. Freshly extracted intact premolars, removed for orthodontic reasons were used. Root canal treatment was performed in each tooth. And then the outer surface and crown portion of the teeth was sealed with wax leaving the CEJ. The prepared teeth mounted on the wax laminates were placed in plastic assay tubes containing 1.5ml bidistilled water with their entire root, including the CEJ, submerged in the solution. The teeth were dividied into four groups. Thermo group : thermocatalytic bleaching with superoxol Walk group: walking bleaching with sodium perborate & superoxol Combi group : combination of thermocatalytic & walking bleaching Dw group : walking bleaching with sodium perborate & water The bleaching procedure was performed three times. The bleaching intervals were at 3 days. The hydrogen peroxide present in the assay system was added to ferrous ammonium sulfate resulting in ferric ion release. Upon the addition of potassium thiocyanate a ferrithiocyanate complex results, which absorbs light at the wavelength of 467nm. The radicular penetration of hydrogen peroxide in the four groups was assessed directly using spectrophotometer. The amount of hydrogen peroxide in the samples tested is determined by comparing them with a standard curve generated by known amounts of hydrogen peroxide. The results were obtained as follows : 1. In all experimental groups except the Dw group showed lower penetration amount in day 4 than day 1, there was statistical importance in the difference (P<0.05). 2. After 3rd treatment, Thermo group showed slightly increased value and narrow distribution. Walk group showed much more penetration amount and widely dispersed value. Value of Combi group showed wide distribution without regard to treatment time, but value of Dw group evenly distributed. 3. Thermo group, Walk group and Dw group showed a tendency of increasing penetration amount with increasing treatment times(P<0.01), but Combi group revealed no statistically important differences. 4. Combi group showed the highest degree of penetration. Walk group showed lower penetration than Combi group. Thermo group & Dw group showed lower than Walk group. 5. Cervical root permeability to hydrogen peroxide varied from 0 to 35 %.
A biological active substance was isolated from the cultured medium of Streptomyces sp. and its biochemical characteristics were investigated. Isolation process of the substance was as follows; the pH of filterate of the cultured medium was adjusted to 3.0 with N-hydrochloric acid and saturated with sodium chloride, then chloroform was added to this filterate in one fifth portions and stirred vigorously. After extracting the active substance with chloroform in 3 stages, the chloroform layer combined and evaporatea after dehydrating with sodium sulfate. The substance was found to be to be toxic to various fresh water fishes; the lethal dose for an average size Pseudorasbora parva T. et. S. was 50ug per ml. In the acidic condition, the toxicity of the substance remained fora long time, while in the alkaline state, the toxicity was decreased very fast. This substance was found to be stable to organic solvents, but labile to heat treatment. The maximal revival time of Pseudorasbora parva T. et. S. was about 20 minutes in 25 ug/ml of the substance solution.
The effect of physical and chemical treatments to reduce staphylococcal phages was investigated. To determine impact of physical treatment on viability of phages, two staphylococcal phages (SAP84 and SAP89) were treated with multiple heat ($55^{\circ}C$ and $60^{\circ}C$) and pH (pH4, 7, 10) conditions. Viability of SAP 84 was dramatically reduced at 60C and SAP 89 was completely inactivated at 60C within 25 min. Overall, the two phages were stable under all the pH conditions tested except for the SAP 89 at pH 10. Treatments, a 10% FAS (Ferrous Ammonium Sulfate) solution and various density of ethanol and sodium hypochlorite were used to reduce the two phages. SAP 84 was unstable in 50% and 70% ethanol. However, SAP 84 and SAP 89 showed high tolerance after exposure to 100 ppm of sodium hypochlorite which is known as an effective sterilizer. As soon as the two phages were treated with 10% FAS, which is used as a virucidal agent, they were inactivated and did not form any plaque. The result of this study provides additional evidence that staphylococcal phages can be controlled by various physicochemical treatments.
Soil dispersion and heavy metal leaching with two heavy metal-contaminated soils were studied to derive the optimal dispersion condition in the course of developing the remedial technology using magnetic separation. The dispersion solutions of pyrophosphate, hexametaphosphate, orthophosphate and sodium dodecylsulfate (SDS) at 1 - 200 mM and the pH of solutions was adjusted to be 9 - 12 with NaOH. The clay content of suspension as an indicator of dispersion rate and the heavy metal concentration of the solution were tested at the different pHs and concentrations of the dispersion solution during the experiment. The dispersion rate increased with increasing the pH and dispersion agent concentration of the solution. The dispersion efficiency of the agents showed as follows: pyrophosphate > hexametaphosphate > SDS > orthophosphate. Arsenic leaching was sharply increased at 50 mM of phosphates and 100 mM of SDS. The adsorption of $OH^-$, phosphates and dodecysulfate on the surface of Fe- and Mn-oxides and soil organic matter and the broken edge of clay mineral might decrease the surface charge and might increase the repulsion force among soil particles. The competition between arsenic and $OH^-$, phosphates and dodecylsulfate for the adsorption site of soil particles might induce the arsenic leaching. The dispersion and heavy metal leaching data indicate that pH 11 and 10 mM pyrophosphate is the optimum dispersion solution for maximizing dispersion and minimizing heavy metal leaching.
Lee, Seung Bum;Jeon, Gil Song;Jung, Rae Yoon;Hong, In Kwon
Applied Chemistry for Engineering
/
v.27
no.1
/
pp.21-25
/
2016
When copper alloy is used in etching process for the production of lead frame, the high concentration of heavy metals, such as iron, nickel and zinc may be included in the etching waste. Those etching waste is classified as a specified one. Therefore a customized design was designed for the purification process of the lead frame etching waste liquid containing high concentrations of heavy metals for the production of an electroplating copper(II) oxide. Since the lead frame etching waste solution contains highly concentrated heavy metal species, an ion exchange method is difficult to remove all heavy metals. In this study, a copper(I) chloride was manufactured by using water solubility difference related to the reduction-oxidation method followed by the reunion of copper(II) chloride using sodium sulfate as an oxidant. The hydrazine was chosen as a reducing agent. The optimum added amount was 1.4 mol per 1.0 mol of copper. In the case of removal of heavy metals by using the combination of reduction-oxidation and ion exchange resin methods, 4.3 ppm of $Fe^{3+}$, 2.4 ppm of $Ni^{2+}$ and 0.78 ppm of $Zn^{2+}$ can be reused as raw materials for electroplating copper(II) oxide when repeated three times.
Cellulolytic mocrooganisms were isolated from the cellulose-cultural properties. Among them, Aspergillus clavatus with the highest cellulase activity was identified by the morphological characteristics and it's enzyme activities were compared on the various cultural conditions. It was found that the induction of carboxymethylcellulase(CMCase), avicelase and salicinase in CMC liquid media showed the highest enzyme acitivity on five day's cultivation at $30^{\circ}C$ and thereafter their activities were gradually decreased with time. After crude extracellular enzymes precipitated with the 70% saturated ammonium sulfate solution were dialyzed with 20 mM acetate buffer pH 6.0, each crude enzyme was examined. The optimal activities of CMCase and avicelase were both found to be at $50^{\circ}C$ and pH 6.0. Their thermal stability was appeared at the $50^{\circ}C$. CMCase and avicelase had the maxima activities with 1.5% and 2.2% substrate concentration, respectively. The concentration of 5 mM $Mg^{++}$ or $Ca^{++}$ was found to have a maximum cellulase activity and its activity was inhibited with more than 5 mM $Cu^{++}$ and $Zn^{++}$ concentration. Cellulase activity was also inhibited sensitively by the inhibitors such as 2-mercaptoethanol, iodine and sodium azide.
De Marqui, Alessandra Bernadete Trovo;Vidotto, Alessandra;Polachini, Giovana Mussi;De Mattos Bellato, Claudia;Cabral, Hamilton;Leopoldino, Andreia Machado;De Gois Filho, Jose Francisco;Fukuyama, Erica Erina;Settanni, Flavio Aurelio Parente;Cury, Patricia Maluf;Bonilla-Rodriguez, Gustavo Orlando;Palma, Mario Sergio;Tajara, Eloiza Helena
BMB Reports
/
v.39
no.2
/
pp.216-222
/
2006
In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.
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