• Journal of Microbiology and Biotechnology
• /
• v.19 no.11
• /
• pp.1482-1489
• /
• 2009

In this study, we investigated the anti-proliferative effect of polysaccharides from Salicornia herbacea on HT-29 human colon cancer cells. Crude polysaccharides from S. herbacea (CS) were prepared by extraction with hot steam water, and fine polysaccharides from S. herbacea (PS) were obtained through further size exclusion chromatography. The anti-proliferative effect of CS and PS were measured using the MTS assay, apoptosis analysis, cell cycle analysis, and RT-PCR. HT-29 cells were treated with CS or PS at different dosages (0.5, 1, 2, 4 mg $ml^{-1}$) for 24 or 48 h. CS and PS inhibited proliferation and stimulated apoptosis of cells in a dose-dependent manner. Flow cytometric analysis after Annexin V-FITC and PI staining revealed that treatment with CS or PS increased total apoptotic death of cells to 24.99% or 91.59%, respectively, in comparison with the control (13.51 %). PS increased early apoptotic death substantially - up to 12 times more than the control. Treatment with CS or PS resulted in a concentration-dependent increase of the G2/M cell population of the cell cycle as determined by flow cytometry. G2/M arrest was induced significantly with the highest concentration (4 mg $ml^{-1}$) of PS. RT-PCR was performed to study the correlation between G2/M arrest and transcription of cell cycle control genes. The anti-proliferative activity of CS and PS was accompanied by inhibition of cyclin B1, and Cdc 2 mRNA. Moreover, both CS and PS induced expression of the p53 tumor suppressor gene and the Cdk inhibitor p21. These results suggest that polysaccharides from S. herbacea have anti-cancer activity in human colon cancer cells.

• Food Science and Biotechnology
• /
• v.16 no.4
• /
• pp.600-604
• /
• 2007

In this study, we first prepared 3 kinds of powders with different molecular masses from the crude protein-bound polysaccharide extract of Agraricus blazei Murill through ultrafiltration, followed by spray-drying. Then, the antitumor activities of the powders were analyzed. Size exclusion chromatography coupled with a multi-angle laser-light-scattering system showed the 3 powders had the following molecular ranges: below 10 kDa (SD-1), 10 to 150 kDa (SD-2), and above 150 kDa (SD-3), representing peak molecular weights of $8.26{\times}10^3,\;9.65{\times}10^4$, and $5.94{\times}10^6\;g/mol$, respectively. All the powders stimulated macrophage RAW264.7 cells to produce nitric oxide, of which SD-2 and SD-3 were superior to the crude extract powder (CP-SD), while SD-1 showed the lowest activity. Similar results were found for their cytotoxicities against human cancer cell lines (A549, MCF-7, and AGS), where the highest activity was obtained with the SD-2 treatment for 72 hr at $1,000\;{\mu}g/mL$. The MCF-7 cell line was less sensitive to the powders than the other cells. From this research we found that ultrafiltration, in combination with spray-drying, is applicable for preparing protein-bound polysaccharide powders with higher antitumor activities.

• Polymer(Korea)
• /
• v.39 no.2
• /
• pp.225-234
• /
• 2015

In this study, we proposed to synthesize thermally stable, soluble and conjugated Schiff base polymer (SbP). For this reason, a specific molecule namely 4,4'-thiodiphenol which has sulfur and oxygen bridge in its structure was used to synthesize bi-functional monomers. Bi-functional amino and carbonyl monomers namely 4,4'-[thio-bis(4,1-phenyleneoxy)] dianiline (DIA) and 4,4'-[thiobis(4,1-phenyleneoxy)]dibenzaldehyde (DIB) were prepared from the elimination reaction of 4,4'-thiodiphenol with 4-iodonitrobenzene and 4-iodobenzaldehyde, respectively. The structures of products were confirmed by elemental analysis, FTIR, $^1H$ NMR and $^{13}C$ NMR techniques. The molecular weight distribution parameters of SbP were determined by size exclusion chromatography (SEC). The synthesized SbP was characterized by solubility tests, TG-DTA and DSC. Also, conductivity values of SbP and SbP-iodine complex were determined from their solid conductivity measurements. The conductivity measurements of doped and undoped SbP were carried out by Keithley 2400 electrometer at room temperature and atmospheric pressure, which were calculated via four-point probe technique. When iodine was used as a doping agent, the conductivity of SbP was observed to be increased. Optical band gap ($E_g$) of SbP was also calculated by using UV-Vis spectroscopy. It should be stressed that SbP was a semiconductor which had a potential in electronic and optoelectronic applications, with fairly low band gap. SbP was found to be thermally stable up to $300^{\circ}C$. The char of SbP was observed 29.86% at $1000^{\circ}C$.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.26 no.1
• /
• pp.47-53
• /
• 2018

Recovery of lactic acid from fermentation broth using chemical precipitation was investigated with various chemicals. Effects of chemical types, mixing speeds, settling duration, and solvent addition were evaluated to improve the recovery rates of lactic acid. Overall, recovery efficiencies increased as the dosage of chemicals increased. Recovery rate of lactic acid by CaO was higher than those of $Ca(OH)_2$ and $CaCO_3$. Recovery of lactic acid increased by 48% under the optimized reaction conditions which included a mixing speed at 180 rpm, a settling duration of 24 h, and addition of ethanol at 25%(v/v). Practical application needs to consider types and concentrations of other organic acids as well as lactic acid. Based upon the results of fluorescence excitation emission matrix (FEEM), size exclusion chromatography (SEC), characteristics of recovered lactic acid were same as that in the fermentation broth.

• Korean Journal of Food Science and Technology
• /
• v.30 no.3
• /
• pp.494-497
• /
• 1998

Deodorization of soybean oil was carried out at a temperature range of $220{\sim}280^{\circ}C$ for 1 or 2hrs. HPSEC(high performance size exclusion chromatography) method was used for the analysis of dimeric acids in deodorized soybean oil. Dimeric acid moieties were produced through the deodorization at $240{\sim}280^{\circ}C$ for 2 hrs. The amount of dimeric acids produced was found to be in the range of 0.36 to 3.39%. Dimeric acids were not detected under the deodorizing condition of $240^{\circ}C,\;2\;hrs\;or\;250^{\circ}C,\;1\;hr$. The soybean oil deodorized in these conditions had good quality by the color and viscosity tests. The best condition of minimizing the formation of dimeric acids in soybean oils was the deodorizing conditions of $240^{\circ}C,\;2\;hrs\;or\;250^{\circ}C,\;1hr$.

• Journal of the Korean Chemical Society
• /
• v.36 no.3
• /
• pp.419-427
• /
• 1992

The performance of homemade thermal field-flow fractionation (ThFFF) and the optimization for separation are investigated in terms of retention parameters and selectivity using polystyrene standards. The ribbonlike channel was constructed by sandwiching two Mylar spacers between two highly polished copper bars which serve as channel walls. The hot bar was heated by two electrical cartridge heaters and the cold bar had holes through the entire length, allowing tap water as coolant to enter. It has been observed that the retention of polystyrenes increases as the temperature gradient increases. And the retention decreases as the cold wall temperature increases from 20 to $45^{\circ}C$. In comparison with size exclusion chromatography, ThFFF is more selective but has relatively low efficiency. It has been found that the plate height is proportional to the flow velocity and good estimates of polydispersities can be made directly from the ThFFF data.

• Korean Chemical Engineering Research
• /
• v.43 no.3
• /
• pp.360-365
• /
• 2005

SMB (simulated moving bed) is a continuous chromatographic process by shifting periodically port position. Binary of mixture, Blue dextran and Orange G, was separated by SMB. These components have unique color individually, that is, Blue dextran is blue and Orange G is orange. It is easy to understand SMB process by observing the shift of color changes in SMB. These components was not adsorbed to stationary phase and isolated by difference of size exclusion factor. Mass transfer coefficient was determined by single pulse test under several flow rate conditions. Operation condition was obtained by standing wave theory and optimized for high purities in extract and raffinate streams. Experiment was performed in open loop 4 zone (2-2-2-2) SMB. There are several advantages in open loop SMB, where extract is product for high purity. It is also easy to control flow rate and monitor experimental state during operation. Experimental, extract and raffinate history is well fitted with simulation results, however, column concentration profile is a little different from simulation results. Purities were 99.5％ for extract and 98.9％ for raffinate and extract and raffinate yields were obtained as 98.9％ and 99.4％ respectively.

• Journal of the Korean Magnetic Resonance Society
• /
• v.21 no.2
• /
• pp.72-77
• /
• 2017

${\beta}-catenin$ is a key signaling protein which regulates cell signaling and gene transcription. Abnormal activation of ${\beta}-catenin$ is linked to many cancers, particularly with colorectal cancers. Although many genetic and biological studies on $Wnt/{\beta}-catenin$ have been reported and structures of the complex between ${\beta}-catenin$ and its diverse binding partners have been published, many of them have focused on armadillo repeat domain of ${\beta}-catenin$. Both N- and C-terminal domains have been suggested to regulate interactions of ${\beta}-catenin$ with other molecules, but still little is known about the C-terminal unstructured domain. To investigate the structure of this domain, construct of C-terminus was designed and structural studies were performed using size exclusion chromatography (SEC), circular dichroism (CD), fluorescence and nuclear magnetic resonance (NMR) spectroscopy. We observed that not only the purified full-length construct but the purified C-terminal construct also dimerizes in solution by SEC, suggesting that this domain involves in dimerization of ${\beta}-catenin$. CD and fluorescence data indicate its flexibility and structural formation in the presence of membrane environments.

• Microbiology and Biotechnology Letters
• /
• v.49 no.2
• /
• pp.181-191
• /
• 2021

The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The BoxBehnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.12
• /
• pp.1905-1911
• /
• 2020

Homoserine dehydrogenase (HSD) catalyzes the reversible conversion of ʟ-aspartate-4-semialdehyde to ʟ-homoserine in the aspartate pathway for the biosynthesis of lysine, methionine, threonine, and isoleucine. HSD has attracted great attention for medical and industrial purposes due to its recognized application in the development of pesticides and is being utilized in the large scale production of ʟ-lysine. In this study, HSD from Bacillus subtilis (BsHSD) was overexpressed in Escherichia coli and purified to homogeneity for biochemical characterization. We examined the enzymatic activity of BsHSD for ʟ-homoserine oxidation and found that BsHSD exclusively prefers NADP+ to NAD+ and that its activity was maximal at pH 9.0 and in the presence of 0.4 M NaCl. By kinetic analysis, Km values for ʟ-homoserine and NADP+ were found to be 35.08 ± 2.91 mM and 0.39 ± 0.05 mM, respectively, and the Vmax values were 2.72 ± 0.06 μmol/min-1 mg-1 and 2.79 ± 0.11 μmol/min-1 mg-1, respectively. The apparent molecular mass determined with size-exclusion chromatography indicated that BsHSD forms a tetramer, in contrast to the previously reported dimeric HSDs from other organisms. This novel oligomeric assembly can be attributed to the additional C-terminal ACT domain of BsHSD. Thermal denaturation monitoring by circular dichroism spectroscopy was used to determine its melting temperature, which was 54.8℃. The molecular and biochemical features of BsHSD revealed in this study may lay the foundation for future studies on amino acid metabolism and its application for industrial and medical purposes.