• Journal of Life Science
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• v.32 no.4
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• pp.303-309
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• 2022

Dystaenia takesimana is an endemic plant found only in Korea, especially on Ulleung Island. The leaves and roots of D. takesimana have been used as food, forage, and oriental medicine. Anti-bacterial, anti-inflammation, antioxidant, and α-glucosidase inhibition biological activities have been reported in the plant's root extract. However, studies concerning the anti-thrombosis activities of D. takesimana are still in the rudimentary stage. In this study, the extracts of the leaf (DT-L), stem (DT-S), and root (DT-R) of D. takesimana were prepared using 70% ethanol, and their anti-thrombosis activities were evaluated. DT-L extracts (0.25 mg/ml) showed strong inhibitions against platelet aggregation, comparable to aspirin, with strong radical scavenging activities. Furthermore, the DT-L extract did not show any RBC hemolysis up to 1 m/ml. The ant-coagulation and antioxidant activities of the DT-S extract were ignorable. While the DT-R extract showed inhibitions against thrombin and blood coagulation factors, it also showed strong platelet aggregation. This is a first report of the anti-thrombosis activities of D. takesimana, and our results suggest that DT-L could be developed as a valuable bioresource for high value-added products.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.197-203
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• 2001

A fusion protein, consisting of human epidermal growth factor as a recognition domain and human angiogenin as a toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably due to the opposite surface charge due to vastly different pI values of each domain. Solid-phase refolding process exploiting ionic interactions between the solid matrix and the protein was tried, but the ionic binding yield was very low regardless of the resins and pH conditions used. To provide higher affinity toward the solid matrix, six lysine residues were tagged to the N -terminus of the hEGF domain When the cation exchange resins such as heparin- or CM-Sepharose were used as the matrix, the adsorption capacity increased 2.5-3 times and the subsequent refolding yield increased nearly IS times compared to the conventional process.

• Korean Journal of Agricultural Science
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• v.41 no.3
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• pp.237-243
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• 2014

Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.4
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• pp.271-276
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• 2012

In this study, the dispersion ability of powder in low viscosity solubilization system that depends on variety and amount of additives and powders was investigated. A PMMA powder shows outstanding dispersion ability because of its repulsive force of partially charged chain and low density of porous structure. A sample, which contains salts, showed better dispersion tendency than a sample without any additives. The dispersion ability was quantity of salts dependent. Furthermore, a sample with divalent ion salts, like $MgSO_4$, showed better dispersion tendency than that of monovalent ion salts, like NaCl or KCl. The reason for the better dispersion tendency was due to the existence of ionized salts around the powders which significantly improves repulsive force between powders and consequently reduces powder aggregation. The sample with chelating agent, like EDTA as an additive, had improved dispersion ability. EDTA chelates and blocks metal cation therefore anion's character is maximized and repulsive force between powders is improved. As a result, salts and EDTA help to improve the powder dispersion ability and the stability of product.

• Journal of Life Science
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• v.25 no.7
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• pp.757-764
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• 2015

The immature fruit of Rubus coreanus Miquel (bokbunja in Korean) is mainly consumed as a fruit wine due to its sour taste and low sugar content. The lees (LBW) remaining after the production of bokbunja wine are discarded as they have no specific usage. The aim of this study was to develop high-value-added biomaterials for functional foods and beauty/health products by investigating the anti-microbial, anti-oxidant, and anti-thrombosis activities of LBW using ethanol and hot water extracts and their subsequent organic solvent fractions. The ethyl acetate (EA) fraction of LBW extracts has a high polyphenol content (413–459 mg/g), and showed strong anti-microbial activity against gram-positive bacteria. The EA fraction also showed excellent radical-scavenging activity against DPPH anion, ABTS cation, and nitrite, with strong reducing power. The polyphenol-enriched EA fraction strongly inhibited thrombin, prothrombin, and blood coagulation factors. The butanol fraction showed a specific inhibition of coagulation factors, as measured in activated partial thromboplastin time assay, which is linked to intrinsic blood coagulation. The butanol fraction also showed strong inhibition of platelet aggregation, at levels comparable to aspirin. The residue of the hot-water extract, which is produced by sequential solvent fractionation of the LBW extract, showed superior inhibition against platelet aggregation when compared to aspirin. Our results suggest that the LBW, which are currently discarded, are a promising source of novel functional foods and beauty/health products.

• Korean Journal of Soil Science and Fertilizer
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• v.17 no.3
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• pp.224-231
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• 1984

Four kinds of soil conditioners, such as red earth, clayey tidal deposit, silty tidal deposit and tertiary deposit were evaluated for the rice growth and for the effectiveness of soil improvement in sandy paddy soil (Gangseo series) whose percolation and nutrient leaching are usually severe. Experiment was conducted at the farmer's field in Hackpo Ri, Bugog Myeon, Changyeong Gun, Gyeong Nam Province for two years (1980-1981) with using two rice varieties, Milyang 42 for 1980 and cheongcheongbyeo for 1981. For both experimental years, the grain yields were increased on the plots that were treated with soil conditioners compared with nontreated plot. The increase in grain yield was particularly high on the plots treated with clayey tidal deposit and teritary deposit. The dry weight of the plant and the uptake amount of inorganic ingredient tended to increase on the plots of clayey tidal deposit and tertiary deposit during whole rice growing season because of the residual effect of soil conditioners in the second experimental year. Cation exchange capacity, available silicate, exchangeable potassium and active iron was increased by the treatment with the soil conditoners. Especially cation exchange capacity was highest in the tertiary deposit treatment plot. Aggregation content and plastic index were increased for the all treatment plots with soil conditioner, however, hydraulic conductivity was conspicuously decreased by the tertiary deposit and clayey tidal deposit treatments.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.311-316
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• 2016

This study was designed to develop the functional beverage using Salicornia herbacea L (SH). To evaluate the anti-oxidation and anti-thrombosis activities of SH, the ethanol extracts of leaf (LF), stem (ST) and seed (SD) of SH were prepared. Components analysis showed the total polyphenol of LF > ST > SD, and the total flavonoid of LF > SE > ST, respectively. Amounts of total sugar and reducing sugar of LF and ST were similar, but the SD had only 1/10 of sugar contents of LF. Anti-oxidation activities of each extract determined by calculated RC₅₀ showed LF > SD > ST for DPPH anion, nitrite and reducing power, and LF = SD > ST for ABTS cation scavenging activity, respectively. In anti-coagulation assay, SD showed strong inhibition activity against thrombin and blood coagulation factors. All the extracts had no platelet aggregation activities against human platelet and no hemolysis against human RBC. Our results suggest that the SD from SH has a great potential as a new anti-coagulation agent.

• Journal of Life Science
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• v.27 no.2
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• pp.131-138
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• 2017

In this study, for the efficient use of the byproduct of the omija (Schizandra chinensis Baillon: SC) processing industry, the ethanol extracts of the fruit (F), seed (S), and pomace (P) of SC were prepared, and their useful bioactivities were evaluated. For F-SC, S-SC, and P-SC, the extraction yields were 28.3%, 22.1%, and 7.2%, respectively, and the polyphenol contents were 8.81, 37.22, and 9.20 mg/g, respectively. The total flavonoid content in P-SC (4.31 mg/g) was 3.5-fold higher than that in F-SC (0.76 mg/g). In an antioxidation activity assay, P-SC showed stronger radical scavenging activities against DPPH anion, ABTS cation, and nitrite and stronger reducing power activities than the other extracts. The calculated concentration required for 50% radical scavenging activity, $RC_{50}s$, of P-SC for DPPH anion, ABTS cation, and nitrite was 226.2, 192.5, and $92.5{\mu}g/ml$, respectively. In an antimicrobial activity assay, F-SC, S-SC, and P-SC showed similarly strong growth inhibitions against Bacillus subtilis and P. vulgaris at a concentration of 0.5 mg/disc. F-SC and P-SC showed 15-fold extended time in thrombin, prothrombin, and activated partial thromboplastin time assays at a concentration of 5 mg/ml. The anticoagulation activity of P-SC (2.5 mg/ml) was comparable to that of aspirin (1.5 mg/ml). Furthermore, F-SC and S-SC showed very good platelet aggregation inhibitory activities. F-SC, S-SC, and P-SC did not show significant hemolysis against human red blood cell up to a concentration of 0.5 mg/ml. These results suggest that S-SC and P-SC, both of which are byproducts of the omija processing industry, show strong potential as novel antioxidant, antimicrobial, and antithrombosis agents.

• KSBB Journal
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• v.17 no.2
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• pp.153-157
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• 2002

A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1749-1759
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• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.