• The Korean Journal of Zoology
• /
• v.33 no.3
• /
• pp.330-336
• /
• 1990

Changes in esterase activity and zymogram pattern during development in Pieirs rapae L. were investigated and three esterases (E2, E6 and E11) from the late 5th instar larvae were purified. Esterase activity in whole body increased rapidly during 5th instar larval stages and reached a peak at the late 5th instar larval stage. The number and intensity of esterase band from whole body and midgut also showed a peak at the late 5th instar larval stage. Purification of esterase was performed using gel filtration on Sephadex G-lOO, ion-exchange chromatography on DEAE-trisacryl and preparative electrophoresis. The final purities of these enzymes were about 30 to 60-fold.

• Journal of Pharmacopuncture
• /
• v.25 no.2
• /
• pp.114-120
• /
• 2022

Objectives: Antivenom serums have been used extensively for over a century and are the only effective treatment option for snake bites and other dangerous animal envenomations. In therapeutic serum centers, a wide range of antivenoms is made from animal serum, mainly equine and sheep, that are immunized with single or multiple venoms. This work aimed to use caprylic acid (CA) to purify therapeutic snake antivenom. Methods: Plasma was obtained from equine immunized with a mixture of venoms. Immunized plasma was obtained by precipitation of different concentrations (2-5%) of CA. This methodology was compared to that based on ammonium sulfate (AS) precipitation. Sediment plasma proteins were purified by ion-exchange chromatography. Protein assay, SDSPAGE, and agar gel diffusion were performed. Results: The total protein precipitation with AS was higher than precipitation with CA, but the best results were obtained when CA was added to the plasma until a final CA concentration of 5% was reached. Chromatography and electrophoresis indicated a stronger band for the 5% CA, and the gel diffusion assay showed antigen-antibody interaction in the purified serum. Conclusion: The use of CA compared to the routine method for purifying hyperimmune serums is a practical and cost-effective method for preparing and producing therapeutic serums. It constitutes a potentially valuable technology for alleviating the critical shortage of antivenom in Iran.

• Journal of Soil and Groundwater Environment
• /
• v.12 no.2
• /
• pp.55-64
• /
• 2007

An analytical method using ion-exchange chromatography was developed for simultaneous quantification of low-molecularweight organic acids ($C_1-C_6$ aliphatic carboxylic acids) and inorganic anions, and then applied to the assessment of ground water contaminated by leachates from a municipal solid waste landfill. Peak interferences of halide ions to organic acids were removed by pretreatment of water samples with Ag-containing cartridges. This method allowed accurate detection of low-molecular weight organic acids (i.e., formate, acetate, propionate, pyruvate, succinate, and oxalateas) low as 0.5 mg/L with a linear dynamic range up to 20 mg/L within 11 min run time along with typical inorganic anions. High level of pyruvate and low level of formate and acetate were detected in groundwater and landfill leachates using the analytical method. Pyruvate concentration in groundwater showed a significant correlation with concentrations of $Cl^-$ and $HCO_3^-$, and pyruvate levels decreased along the downgradient from the landfill, indicating the sources of pyruvate are landfill leachate.

• Applied Biological Chemistry
• /
• v.12
• /
• pp.43-51
• /
• 1969

The purpose of this study was to determine protein amino acid contents of some Korean foods by gas-liquid chromatography, and to evaluate this technique as a procedure for the quantitative determination of amino acids in foods. The crude protein content of foods was also estimated from the nitrogen content. 1. Nitrogen content of each food sample was determined previously to adjust the amount of sample for GLC analysis 2. In the analysis of 17 known amino acids, a linear relationship was found between the weight of 13 amino acids of 17 amino acids, the internal standard as well as the injection volume of a mixture and the detector responses for the derivatives of the amino acids. No response for arginine, cystein, histidine, and tyrosine was observed. 3. The relative molar response (RMR) values for the 13 amino acids of standard solution relative to glutamic acid as '1.00' were obtained under normal operating conditions with a hydrogen flame ionization detector. 4. The recovery of amino acids from their mixtures with natural food materials was carried out. The recoveries were essentially quantitative except threonine and serine. An overall mean recovery of 11 amino acids was $101.4{\pm}8.4$ per cent before hydrolysis and $98.1{\pm}8.7$ per cent after hydrolysis of samples. 5. The comparative analysis of the acid hydrolysates of two food samples by gas-liquid and ion-exchange chromatographic analysis were carried out. In white-bait pemmican, only threonine and asparagine amounts by GLC analysis had similar values to those obtained by ion-exchange chromatography. The other seven amino acids gave higher values as measured by GLC than by ion-exchange. With the food sample, soybean, alanine, valine, asparagine, and glutamic acid were in good agreement in two analysis, while leucine, proline, threonine, phenylalanine, and lysine were found in slightly higher concentrations in the GLC analysis. 6. Grant variations of amino acid content were found among samples analyzed. The amino acid contents of each sample were compared with the values found in the literature.

• Food Science and Preservation
• /
• v.10 no.2
• /
• pp.246-251
• /
• 2003

Pretense produced by Bacillus subtilis PANH765 was purified from culture supernatant by using ammonium sulfate fractionation DEAE-cellulose ion exchange chromatography, and gel filtration with Sephacryl S 200 HR and Sepharose CL-6B. DEAE-cellulose ion exchange column chromatography, separated the pretense into one fraction. This fraction was further purified using Sephacryl S 200 HR and Sepharose CL-6B gel titration. The molecular mass of pretense was estimated to be 35.0 kDa by the SDS-PAGE and gel filtration using Sepharose CL-6B. The results indicated that the purified pretense are monomeric proteins. Specific activity and purification folds of pretense were 657 U/mg and 4.35, respectively. The optimum temperature, optimum pit stable at a temperature range and pH ranges for the purified protease were 65$^{\circ}C$, 7.05, 50 ∼ 75$^{\circ}C$ and 6.0 ∼ 7.5, respectively. The pretense activity was decreased by the presence of PMSF and DFP, which the protease activity was increased by the presence of Na$\^$+/, K$\^$+/, Mg$\^$2＋/ and NH$_4$$\^$+/ ions.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.10 no.3
• /
• pp.236-241
• /
• 2005

A decolorization process using ion exchange chromatography was developed to refine rhEGF as a cosmetic ingredient. A macroreticular resin (D314) was selected, with respect to its high decolorization rate and recovery yield of rhEGF, and the operational conditions of the decolorization process optimized. The optimum conditions were as follows: the rhEGF effluent was ion exchanged at a flow rate of 60.0mL/h, with an effluent pH 5.0, using a chromatographic column (i.d. 16mm) packed with D314, with a 7.5cm in bed height. The decolorization process was carried out under the optimum conditions, and halted when the effluent volume reached 350mL, giving a decolorization rate and recovery yield of rhEGF higher than 67 and $80\%$, respectively. When the decolorization rate exceeded $67\%$, the final product turned out to be white or light yellowish, which was to the satisfaction of the cosmetic standard.

• Korean Journal of Microbiology
• /
• v.27 no.2
• /
• pp.117-123
• /
• 1989

Radioactive N-$\alpha$-p-nitrobenzoxycarbonyl (NBZ)-L-[2,$3-^{3}$H] arginyl diazomethane was used as an affinity label for the vacuolar arginine transporter in Neurospora crassa. Vacuolar matrix proteins were removed by fracturing the membranes with freeze-thaw method in dry ice/ethanol bath. Vacuolar membrane proteins were then wasged with 500mM NaCl to remove ionically bound derivatives and peripheral membrane proteins from vacuolar membranes. After dissolved in 1% Titon X-100, dissolved vacuolar memvrane proteins were separated with molecular sieve column chromatography, anion and cation exchange chromatographies. The arginine transporter was purified giving the purification factor of 1136.

• Parasites, Hosts and Diseases
• /
• v.30 no.3
• /
• pp.191-200
• /
• 1992

A proteolytic enzyme was purified from the tissue extract of spargana (plerocercoids of Spirometra erinacei) by DEAE-Trisacryl M ion exchange chromatography and thiopropyl-sepharose affinity chromatography resulted in a 21-fold purification. The proteinase activity was assayed with a synthetic fluorescent substrate, carbobensoxy-phenylalanyl-7-amiso-4-trifluoromethyl-coumarin. SDS-polyacplamide gel electrophoresis of the purified materials revealed a single 28,000 dalton band. Inhibitor profiles of the band indicated that it belonged to cysteine endopeptidases. It exhibited identical pH curves with optimum at pH 5,5, and 50% activity from pH 4.7 to 8. It could completely degrade collagen chains to three identical products. It also showed some activity on hemoglobin. Furthermore, the band on immunoblots was reactive to the sera of sparganosis patients. These results suggest that the proteolytic enzyme belongs to cysteine proteinase which plays a role in the tissue penetration. Also it may be used as the antigen for diagnosis of active sparganosis.

• Journal of Dairy Science and Biotechnology
• /
• v.23 no.1
• /
• pp.1-8
• /
• 2005

The purpose of this study was to demonstrate biochemical properties and antibacterial activity of lactoferrin(Lf) obtained from the colostrum of Korean native cow. Lactoferrin was isolated from the colostrum of Korean native cow by purification steps using batch extraction, ion exchange chromatography, gel filtration chromatography, affinity chromatography. Other whey protein components that is similar molecular weight and affinity to lactoferrin were gradually removed from crude Korean Native cow's lactoferrin during the purification steps. The molecular weight of the purified Korean native cow's Lf(K-Lf) was 81 kDa, the isoelectric point was 9, and the content of iron was 0.56mg/g, which is indicated that iron saturation of the K-Lf was 40.6%. Amino acid composition and a-helix content were different K-Lf from bovine Lf(B-Lf). Antibacterial activity of E. coli O111 by K-Lf was lower than that of B-Lf. A minimal inhibitory concentration(MIC) of K-Lf and B-Lf was 2.75mg/ml and 1.5mg/ml respectively.

• Membrane Journal
• /
• v.31 no.2
• /
• pp.145-152
• /
• 2021

While there are increasing demands on biomolecules separation, resin chromatography lacks in terms of throughput and membrane chromatography is an alternative with high binding capacity and enhanced mass transfer properties. Unlike typical membrane processing, where the performance can only be empirically assessed, understanding how mechanisms work in membrane chromatography is decisive to design biospecific processing. This short review covers three separation mechanisms, including affinity interaction modes for selectively capturing bulk molecules using biospecific sites, ion exchange modes for binding biomolecules using net charges and hydrophobic interaction modes for binding targeted, hydrophobic species. The parameters in designing membrane chromatography that should be considered operation-wise or material-wise, are also further detailed in this paper.