• 한국생물공학회:학술대회논문집
• /
• 2000.04a
• /
• pp.501-504
• /
• 2000

The purification of a thermostable esterase expressed in Escherichia coli was investigated using thermoprecipitation of unclarified cell homogenates followed by after applying the heat-treated lysate to phenyl-sepharose column, and elution with detergent. Heat treatment at $70^{cdot}C$ was capable of removing to E. coli proteins. Specially, the thermoprecipitation with 15% polyethylene glycol 8000 can remove host proteins and nucleic acids efficiently. Various detergents were used to recover the esterase, which was strongly bound to phenyl-sepharose resin. Triton X-100, non-ionic detergent, was found to be the most efficient of all tested detergents.

• Journal of Microbiology
• /
• v.40 no.1
• /
• pp.26-32
• /
• 2002

From the culture supernatant of the psychrotrophic strain of Bacillus amyloliquefaciens an extracellular serine protease was purified to apparent homogeneity by successive purification steps using QAE-Sephadex, SP-Sephadex and Sephacryl S-100 column chromatography. The pretense is monomeric, with a relative molecular mass of 23,000. It is inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride, but not by EDTA. The enzyme is most active at pH 9-10 and at $45^{\circ}C$, although it is unstable at $60^{\circ}C$.

• The Korean Journal of Food And Nutrition
• /
• v.13 no.2
• /
• pp.146-151
• /
• 2000

To evaluate anticaries and antiinflammation of Aloe vera peel, antimicrobial substances were extracted from Aloe vera peel and identified. The antimicrobial active substances of water extract were successfully purified with solvent fractionation, silica gel column chromatography, preparative thin layer chromatography and UV spectrophotometer. Two purified active substances were identified as aloe-emodin and barbaloin by Mass Spectrometer, 1H-NMR and FT-IR.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.4
• /
• pp.719-725
• /
• 2000

This study focused on the purification and characterization of ACE inhibitor from Porphyra yezoensis. The dried Porphyra yezoensis was ground and hydrolyzed with 2.5 N HCl, followed by neutralization and centrifugation. Then, the subsequential purification of ACE inhibitor was carried out by Amberlite XAD 8, DEAE-Toyopearl 650C, Sephadex LH-20 column chromatography and reverse phase HPLC with C18 column. The purified ACE inhibitor was peptide which consisted of glycine (24.5%), arginine (56.8%) and proline (18.8%). Also, it showed the competitive inhibition pattern to ACE. The apparent molecular mass of purified peptide was 580 dalton, and an IC50 value of ACE inhibitor was 10.6 $\mu\textrm{g}$.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.1
• /
• pp.108-112
• /
• 2009

A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

• Journal of Life Science
• /
• v.19 no.8
• /
• pp.1125-1131
• /
• 2009

The antimicrobial activities of Helicobacter pylori as a functional food source with water and 60% ethanol extracts from Rododendron mucronulatum Turcz. flowers were examined. The total phenol content of 60% ethanol extracts (30.6${\pm}$0.14 mg/g) from Rododendron mucronulatum Turcz. flowers was higher than that of water extracts (23.2${\pm}$0.21 mg/g). The inhibitory activities of Rododendron mucronulatum Turcz. extracts on H. pylori was determined to clear zone of 15 mm in 80% ethanol extracts. Purification of inhibitory compounds was carried out in Sephadex LH-20 and MCI-gel CHP-20 column chromatography using a gradient procedure, with increasing ethanol(0${\rightarrow}$100%) in $H_2O$. The chemical structure of the purified inhibitory compounds of H. pylori was identified to be quercitrin (quercetin-3-O- rhamnopyranoside), myricitrin (myricetin-3-O-rhamnopyranoside), quercetin by FAB-MS, NMR and IR spectra.

• Korean Journal of Food Science and Technology
• /
• v.36 no.2
• /
• pp.361-364
• /
• 2004

EPA and DHA extracted from methyl esterified squid oil were purified by silver exchanged resin, silver nitrate-impregnated silica gel, silver exchanged zeolite, and silica gel column chromatography, among which column chromatography using mixture of silver exchanged resin and silica gel (10% by weight) showed the best result. By this simple purification method, EPA and DHA were concentrated from 12.5 to 27.9% (yield, 86,0%) and from 21.7 to 49.5% (yield, 87.3%), respectively. Silver exchanged resin had additional advantages of outstanding reusability and simple recovery of silver.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.4
• /
• pp.562-566
• /
• 2003

A $\beta$-mannanase of Bacillus sp. was purified by DEAE Sephacel ion exchange column chromatography. The specific activity of the purified enzyme was 17.41 units/mg protein, representing an 84.74-folds purification of the original crude extract. For the separation of two types of hydrolysates by the action of purified $\beta$-mannanase, carbon column chromatography, sephadex G-25 column chromatography and thin layer chromatography were accomplished. Main hydrolysates were D.P value 5 and 7 containing of low D.P values. By the method of FACE (Fluorophore Assisted Carbohydrate Electrophoresis), two types of hydrolysates were identified to homo type.

• Korean Journal of Food Science and Technology
• /
• v.19 no.6
• /
• pp.506-514
• /
• 1987

Peroxidase was purified to a homogeneous state from Castanea Semen by ammonium sulfate precipitation, DEAE-cellulose column chromatography, gel filtration on sephadex G-100 and HPLC, and the purification fold was 65.3. The molecular weight of the enzyme was estimated to be about 35,000 by HPLC. In properties of the enzyme which was purified up to sephadex G-100 column chromatography, the optimum pH and temperature were 5.0 and $50^{\circ}C$, respectively. By heating the enzyme at $80^{\circ}C$ for 1.73 min., the enzyme activity was decreased to 10%. The enzyme was active toward aromatic amines such as o-phenylenediamine and p-phenylendiamine. Kinetic studies indicated a Km of 2.6mM for o-phenylenediamine at an optimal hydrogen-peroxide concentration and a Km of 10mM for hydrogenperoxide at an optimal o-phenylenediamine concentration. Among the reagents tested, L-ascorbic acid and sodium L-ascorbate inhibited significantly the enzyme, while $Ca^{++}$ and $Ba^{++}$ activated the enzyme at the concentration of 1mM and 5mM.

• Journal of the Korean Institute of Gas
• /
• v.10 no.2 s.31
• /
• pp.22-26
• /
• 2006

In this study, modeling and simulation works using Aspen Plus were carried out for DME separation and purification process of pilot plant for the daily production of 50 kg of DME. For modeling of the entire DME separation unit, NRTL liquid activity coefficient model was used for the prediction of liquid phase non-idealities, Henry's law option was also used for the estimation of solubilities of light gases in solvents and SRK equation of state model was utilized for the description of vapor phase non-idealities. DME having over 98 wt% purity was obtained as a side distillate product in a DME purification column.