• Membrane Journal
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• v.19 no.2
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• pp.113-121
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• 2009

Porous affinity chitosan and chitin membranes with good mechanical strength and high protein binding capacity were prepared by using silica particles as porogen. The maximum binding capacity of affinity chitosan membrane for BSA protein is 21.8mg/mL, and that of affinity chitin membrane for lysozyme enzyme is 26.1mg/mL. Chromatographic separations of BSA and lysozyme proteins using the porous affinity chitosan and chitin membranes were performed with change of the flow rate, loading amount and concentration of protein loading solutions. Protein eluted amount and binding yield were calculated from the filtration chromatograms consisted of loading/washing/elution sequences. Protein binding amount and yield were increased with decreasing of flow rate, increasing of loading amount and concentration of protein loading solutions. Those results suggest that the porous chitosan and chitin membranes prepared by using silica particles as porogen are suitable in affinity filtration chromatography for large scale separation of proteins.

• KSBB Journal
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• v.18 no.6
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• pp.500-505
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• 2003

‘LK (lipoprotein kringle) 68’is a polypeptide of a modified ansiostatin consisting of three kringle structures that might be clinically useful as a potential cancer therapeutics. It can be produced by overexpressing it as inclusion body in recombinant E. coli. In this study, solid-phase refolding processes using packed bed adsorption (PBA) and expanded bed adsorption (EBA) column were carried out to compare their refolding yields with that of the conventional, solution-phase refolding process, For the solution-phase and the PBA-mediated processes employing Q-Sepharose, washed inclusion body was used as the starting material, whereas both washed inclusion body and E. coli homogenate were used for the EBA-mediated process employing streamline DEAE. On the final recovery LK68 per unit mass of wet cell basis, the EBA- and PBA-mediated processes showed about 2.7- and 1.5-fold higher yields, respectively, than the solution-phase refolding method. The solid-phase refolded LK68 demonstrated the same Iysine binding bioactivity and the retention time in the RP-and SEC-HPLC as those of the native protein.

• Journal of the Korean Chemical Society
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• v.38 no.2
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• pp.151-159
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• 1994

The portable gas chromatography system was developed which was consisted of ambient vapor sampler(AVS), short capillary column(3 m long, 0.32 mm i.d. GC(SCCGC), photoionization detector (PID) and vacuum pump which was operated at subambient pressure. The seletion of capillary column was based on the theoretical calculation from Golay equation. The pressure ratio of column inlet and outlet appropriated between 1.03 and 1.2 in the system. The available column flow were 0.87∼4.63 ml/min at the pressure ratios. The AVS consisted of three concentric tubes and enables rapid, repetitive introduction of vapor samples directly into capillary column and showed good reproducibility. The subambient column outlet pressure with PID resulted in a significant increase in the optimum column flow, permitting rapid analysis. The baseline separation of m-xylene and o-xylene was able to within 40 second with the system. Parameters affecting the column resolving power were sampling duration, column length and diameter, and the pressure ratio. Effects of these parameters were investigated using bezene derivative compounds.

• Journal of Plant Biology
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• v.39 no.1
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• pp.41-48
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• 1996

Two forms of homo serine dehydrogenase have been isolated from 8-day-old cotyledons of Canavalin lineata by a heat denaturation, ammonium sulfate fractionation, DEAE-8ephacel ion exchange and Sephacryl 8-300 gel filtration chromatographies, and Pro cion red dye, Cibacron blue dye and Resource Q column chromatographies. The molecular weights of T -form (threonine-sensitive) and K-form(threonine- insensitive) were estimated to 230 kD and 135 kD, respectively. In the presence of 10 mM threonine, the activity of T-form was inhibited with almost 70%, but that of K-form was not at all. The Km values tor homo serine of T- and Kform were 1.6 mM and 0.3 mM, respectively. The Km values for NAD of T- and K-form were 2.34 mM and 0.03 mM, respectively. And Km values for NADP of two isozymes were the same as 0.01 mM. The activities of T- and K-form were markedly stimulated up to 4.9and 2.8-fold, respectively, by 400 mM KCI. The partial purified(gel filtration) enzymes(Tform and K-form) can be reversibly converted.verted.

• KSBB Journal
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• v.19 no.4
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• pp.263-268
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• 2004

The separation method using chiral stationary phase in preparation of chiral compound was wildly used, but in this work, supercritical fluid chromatography was suggested in the stability to resolve the chiral mixtures. To determine the optimum operating condition of the racemic ibuprofen, the retention factor and resolution with change in pressures, temperatures and the contents of IPA % (vol.) in CO$_2$ were investigated. The retention factor was decreased with increase in pressure and decrease in temperature. The factor was also influenced by the content of IPA in mobile phase, while the resolution was worse with a increase in IPA %. From the experimental results, the desirable separation condition was 130 bar, 311.15 K and 4% IPA in CO$_2$. Compared to the asymmetric peak shape by liquid chromatography, that of supercritical fluid chromatography was symmetric which was a favorable condition for preparative separation.

• Analytical Science and Technology
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• v.11 no.4
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• pp.292-296
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• 1998

Normal phase or reversed phase liquid chromatographic separation of isoquinoline of heterocyclic compounds and structural isomers of external substituents, $COOCH_3$, CN and $CH_3$ has been carried out by using several different columns and various mobile phases. From this results, the order of elution of heterocyclic compounds appears to depend on the solvent effect with kinds of mobile phases. Retention mechanism of normal phase system for 2-methylindoline, 2-methylindole, benzoxazole and benzothiazole was also studied depending on adsorption strength between solute and stationary phase of column. However, retention factors of reversed phase system were found on hydrophobic interaction with solvophobic effect.

• Analytical Science and Technology
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• v.5 no.2
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• pp.153-158
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• 1992

In this work, we developed supercritical fluid chromatographic methods for the samples which are difficult to analyze with conventional GC or HPLC. Long-chain Hydrocarbons, mink oils and soybean oils unsuitable for GC because of their low volatility or limited thermal stability were separated by SFC due to the high solvating properties of supercritical carbon dioxide fluids. In our research, a new method for the analysis of polar fatty acids and pesticides was developed. This method should be used to overcome problems with polar samples in SFC.

• Korean Chemical Engineering Research
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• v.44 no.6
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• pp.555-562
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• 2006

Recently, liquid chromatography (LC) has been used more frequently to separate drugs and natural substances. Especially, to selection of the solutes from the products, the operation condition of analytical chromatography should be necessarily determined. So accurate computer modeling and simulation of chromatographic performances has become a necessary part of the development and design of processes. High-Purity Separation Lab. Inha University developed the resulting HCI software for the purpose of the optimization of chromatographic performances. The HCI program was utilized to find the optimum operating condition more accurately and rapidly, reducing the number of many possible experiments. The elution profiles were calculated by the plate theory based on the three retention mechanism of capacity factor.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.8
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• pp.1323-1327
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• 2003

Persimmon has been considered to have therapeutic values for various diseases in Korea. Dried persimmon has been applied to wounded parts for anti-inflammatory and analgesic activities. Anti-coagulant fraction from Persimmon stem was purified through gel filtration, phenyl Sepharose, DEAE-Sephadex and additional gel filtration column chromatographies. Its molecular weight was estimated to be 130,000 ∼ 180,000. By element analysis, its main components were C, H, and O. The anti -coagulant was heat- stable and completely inhibited after periodate oxidation, indicating that it was a complex carbohydrate.

• Korean Journal of Microbiology
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• v.43 no.2
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• pp.83-90
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• 2007

Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODI, II, III, IV, V) which can be separated by DEAE-Sepharose chromatography, and SOD III was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD III was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD III was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD III, inhibitory effects of $NaN_{3},\;H_{2}O_{2},\;KCN$ were examined. 10 mM $NaN_{3}$ was able to inhibit 56% of the SOD III activity, which indicates that this SOD is Mn type. The optimum pH of the SOD III was 7.0 and the optimum temperature was $20^{\circ}C$. N-terminal amino acid sequence of purified SOD III was most similar to those of Psudomonase ovalis and Vibrio cholerae among bacteria.