• KSBB Journal
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• v.18 no.1
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• pp.55-61
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• 2003

To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.

• KSBB Journal
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• v.23 no.6
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• pp.513-518
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• 2008

This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

• Korean Journal of Environmental Agriculture
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• v.9 no.2
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• pp.133-141
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• 1990

For the purpose of evaluating nutrient loadings into rivers and lakes from agricultural land, especially from paddy fields and also nutrient degradation in drainage channels, the Total Kjeldahl Nitrogen(TKN) and the Total Phosphorus(TP) were investigated in 29.5 ha. paddy fields in Hwa-Sung, Kyong-Ki, Korea, during the period from May 8, 1989 to Sep. 27, 1989. The results of the study can be su㎜arized as follows : 1. Annual inputs into paddy fields were 180 N-kg/ha 46 P-kg/ha. by fertilization, and 15.0 TKN-kg/ha. 10.0 TP-kg/ha. by irrigation, 8.0 TKN-kg/ha. 0.34 TP-kg/ha. by rainfall respectively. The amount of nutrient involved in surface runoff from paddies was 39.0 TKN-kg/ha. 9.2 TP-kg/ha. and in seepage 7.5 TKN-kg/ha. 2.1 TP-kg/ha. respectively 2. In WS1 stream(reach length equals 950m), nutrients decreased 0.31 TKN-mg/L/km, 0.01 TP-mg/L/km and in WS2 stream (reach length equals 750m) which are more meandering and undulating than WS1, the nutrients decreased 0.84 TKN-mg/L/km, 0.11 TP-mg/L/km. From these results, it was concluded that low stream velocity due to meandering and undulation promotes more degradation of nutrient concentrations. 3. For the purpose of decreasing nutrient loads from paddy fields, the amount of fertilizer used needs to be controlled, irrigation weirs need to be constructed in the drainage channels to delay the transportation of nutrients by decelerating the stream velocity and plants such as plantain-lily need to be cultivated in the channel to consume nutrients and therefore enlarge chances of self-purification.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.57-65
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• 2012

Antifungal compounds from Bacillus polyfermenticus CJ6 were purified using SPE, preparative HPLC, and reverse phase-HPLC. Antifungal compounds from B. polyfermenticus CJ6 were separated into three fractions (8, B, C) using preparative HPLC. LC/MS analysis of antifungal peaks suggested that B. polyfermenticus CJ6 produces lipopeptides; two kinds of iturin A ($C_{14}$, $C_{15}$), three kinds of surfactins ($C_{13}$, $C_{14}$, $C_{15}$), four kinds of fengycin A ($C_{14}$, $C_{15}$, $C_{16}$, $C_{17}$) and two kinds fengycin B ($C_{16}$, $C_{17}$). The antifungal activity of fraction 8, which was presumed as inturin A, was found to be stable after the pH, heat or proteolytic enzyme treatment, but it was unstable at 50-$70^{\circ}C$ for 24 hr. The antifungal activity of fraction B, which presumed as surfactins and fengycin A, was found to be stable after the heat treatment, but it was unstable in the pH 3.0 and after the protease (type I) or ${\alpha}$-chymotrypsin treatment. The antifungal activity of fraction C, which was presumed as fengycin A and B, was found to be stable in the pH 3.0-9.0 range and the heat treatment, but it was unstable with the treatment of protease (type I). The amino acid composition of the purified peaks 8-1 and 8-2 were Asx, Tyr, Gln, Pro, and Ser in a molar ratio of 3:1:1:1:1, which showed the same amino acid composition as iturin. From these results, we confirmed that antifungal compounds from B. polyfermenticus CJ6 most likely belonged to iturin A as well as surfactins and fengycins. As lipopeptides are known to act in a synergistic manner, the antifungal compounds from B. polyfermenticus CJ6 might have potential uses in biotechnology and biopharmaceutical applications.

• Journal of the Korean Society for Marine Environment & Energy
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• v.13 no.1
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• pp.18-29
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• 2010

Offshore subsurface storage of $CO_2$ is regarded as one of the most promising options to response severe climate change. Marine geological storage of $CO_2$ is to capture $CO_2$ from major point sources, to transport to the storage sites and to store $CO_2$ into the offshore subsurface geological structure such as the depleted gas reservoir and deep sea saline aquifer. Since 2005, we have developed relevant technologies for marine geological storage of $CO_2$. Those technologies include possible storage site surveys and basic designs for $CO_2$ transport and storage processes. To design a reliable $CO_2$ marine geological storage system, we devised a hypothetical scenario and used a numerical simulation tool to study its detailed processes. The process of transport $CO_2$ from the onshore capture sites to the offshore storage sites can be simulated with a thermodynamic equation of state. Before going to main calculation of process design, we compared and analyzed the relevant equation of states. To evaluate the predictive accuracies of the examined equation of states, we compare the results of numerical calculations with experimental reference data. Up to now, process design for this $CO_2$ marine geological storage has been carried out mainly on pure $CO_2$. Unfortunately the captured $CO_2$ mixture contains many impurities such as $N_2$, $O_2$, Ar, $H_{2}O$, $SO_{\chi}$, $H_{2}S$. A small amount of impurities can change the thermodynamic properties and then significantly affect the compression, purification and transport processes. This paper analyzes the major design parameters that are useful for constructing onshore and offshore $CO_2$ transport systems. On the basis of a parametric study of the hypothetical scenario, we suggest relevant variation ranges for the design parameters, particularly the flow rate, diameter, temperature, and pressure.

• Journal of Soil and Groundwater Environment
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• v.9 no.4
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• pp.42-51
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• 2004

Peat humin(p-Humin), humic acid(p-HA) and fulvic acid(p-FA) were isolated from Canadian Sphagnum peat moss by dissolution in 0.1M NaOH followed by acid precipitation. After purification cycles, they are characterized for their elemental compositions and, acid/base properties. Functionalities and carbon structures of the humic fractions were also characterized using FT-IR and solid state $^{13}C$-NMR spectroscopy. Those results are compared with one another and with soil humic substances from literatures. Main purpose of this study was to present a chemical and spectroscopic characterization data of humic substance from peat moss needed to evaluate its environmental applicability. The relative proportions of the p-Humin, p-HA and p-FA in the peat moss was $76\%,\;18\%,\;and\;3\%$, respectively, based on the total organic matter content ($957{\pm}32\;g/kg$). Elemental composition of p-Humin were found to be $C_{1.00}H_{1.52}O_{0.79}N_{0.01}$ and had higher H/C and (N+O)/C ratio compared to those of p-HA($C_{1.00}H_{1.09}O_{0.51}N_{0.02}$) and p-FA($C_{1.00}H_{1.08}O_{0.65}N_{0.01}$). Based on the analysis of pH titration data, there are two different types of acidic functional groups in the peat moss and its humic fractions and their proton exchange capacities(PEC, meq/g) were in the order p-FA(4.91) >p-HA(4.09) >p-Humin(2.38). IR spectroscopic results showed that the functionalities of the peat moss humic molecules are similar to those of soil humic substances, and carboxylic acid(-COOH) is main function group providing metal binding sites for Cd(II) sorption. Spectral features obtained from $^{13}C$-NMR indicated that peat moss humic molecules have rather lower degree of humification, and that important structural differences exist between p-Humin and soluble humic fractions(p-HA and p-FA).

• Journal of Wetlands Research
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• v.8 no.2
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• pp.53-64
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• 2006

A study on the restoration process of a stream ecosystem and the water quality renovation technique by removing algae, vegetation and fish monitoring as evaluating the removal of the algae by dietetic characteristics of fishes were performed on Sangdong stream in the B city after stream restoration it to the artificial stream as the cases, restoring urban stream into close-to-nature stream are being increased domestically with the aim of ecological city. As a result, restoration and rehabilitation of the fundamental stream ecosystem was well maintained 4 years later the reclamation at the moment and total 93 diagnosis which were all vascular plant phylum including 44 families, 73 genuses, 79 species and 14 varieties in flora and vegetation community were observed. 3 families, 8 species and 354 populations in total among Fishes were found and Pseudorasbora Parva, Cyprinus Carpic and Carassius Auratus strongly resistant to water pollution were dominantly appeared in order of 50.5% of Pseudorasbora Parva 21.2% of Cyprinus Carpic, 20.9% of Carassius Auratus, 7.1% of Macropodus chinensis and 0.3% of Misqurnus anguillicaudatus according to relative richness index. It turned out to be that Cyprinus Carpic ingests algae over 90% and Carassius Auratus takes it over 30% according to the analysis about the alimentary object of the fishes as a consequences of algae's excrescent from characteristics of the tested experimental stream. It is reported that a Cyprinus Carpic, about 34 cm in length, ingested wet-weight 43.2g algae on the rough analysis toward the sample which makes us recognize how effective a macro community Cyprinus Carpic is for removing algae.As a consequence of this research, the effect of stream ecosystem characteristics and water quality purification could not be expected by aquatic plants and trees which were eliminated at experimental stream. From now on, a close-to-nature stream should be formed of ecological hydraulic and hydrologic engineered modeling from the beginning so that it can perform the water quality purifying function. It is determined that the structure of food chain will be abundantly influenced by the induction of oversized macro community like Cyprinus Carpic because a biomass of a consumer of higher order is increased. It is estimated that the removal algae by fishes is not effective despite in some cases of dietetic characteristics so much more studies should be executed in the future.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.7-17
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• 1985

Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

• Korean Journal of Microbiology
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• v.38 no.3
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• pp.153-161
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• 2002

Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.961-967
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• 2005

Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.