• Journal of Life Science
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• 제9권2호
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• pp.62-66
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• 1999

To overexpress E. coli ADA in host strain E. coli M15, the expression plasmid pQEADD was constructed. To analyze the expression characteristics of ADA, a time course of the expression was first examined. The protein was not detected in no inducted in no induction samples. After the addition of IPTG, a band corresponding to the expected size of ADA was appeared. Its molecular weight was about 36,000 dalton. Maximum expression level was revealed when the cell cultured for 3∼4hrs after induction. This result indicated that the efficient expression of add can be achieved by induction at early logarithmic phase. The effect of different IPTG concentration on the degree of ADA expression was investigated. The expression levels of add were not largely affected by IPTG concentration. Location of overexpression ADA was checked out. A protein band corresponding to the ADA was seen in only crude extract B(insoluble protein). This result suggests that ADA is in E. coli M15. The molecular weight of ADA estimated by SDS-PAGE was approximately 36,000 Da.

• Archives of Pharmacal Research
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• 제25권3호
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• pp.381-386
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• 2002

The objective of this study was to link conformational changes of proteins at a water/methylene chloride interface to their destabilization upon emulsification. When 4 aqueous protein solutions (bovine serum albumin, $\beta$-lactoglobulin, ovalbumin, or ribonuclease) were emulsified in methylene chloride, considerable proportions of all the proteins became water insoluble aggregates. There were also noticeable changes in the compositions of their water-soluble species. A series of water/methylene chloride interfacial reactions upon the proteins was considered a major cause of the phenomena observed. Based on this supposition, the interfacial tension was determined by a Kruss DVT-10 drop volume tensiometer under various experimental conditions. It substantiated that the interfacial tension was high enough to cause the adsorption of all the proteins. Under our experimental conditions, their presence in the aqueous phase resulted in reductions of the interfacial tension by the degrees of 8.5 - 17.1 mN $m^{-1}$. In addition, dynamic changes in the interfacial tension were monitored to compare relative rates at which the adsorbed proteins underwent conformational, structural rearrangements at the interface. Such information helped make a prediction about how easily proteins would denature and aggregate during emulsification. Our study indicated that emulsifying aqueous protein solutions in organic solvents should be handled with care, due to adverse interfacial effects.

• Preventive Nutrition and Food Science
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• 제11권2호
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• pp.100-104
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• 2006

The in vitro antimicrobial and antioxidant activities of the essential oil and methanol extracts of Ledum palustre L. were investigated. Using GC-MS analysis, we identified 37 compounds in the essential oil, constituting 87.58% of the total oil. There are several monoterpenes, of which sabinene is the major compound ($16{\sim}17%$). There are several oxygenated monoterpenes of which terpinen-4-ol(7.6%) and myrtenal (3.5%) are the main constituents. $\beta$-Selinene, a-selinene, $\gamma$-elemene, a-caryophyllene are the main sesquiterpenes ($2{\sim}6%$ range). The oil strongly reduced the diphenylpicrylhydrazyl radical ($IC_{50}=1.56{\mu}g/mL$) formation and exhibited a hydroxyl radical scavenging effect in the $Fe^{3+}-EDTA-H_2O_2$ deoxyribose system ($IC_{50}=2.7{\mu}g/mL$), and also inhibited the nonenzymatic lipid peroxidation of rat liver homogenate ($IC_{50}=13.5{\mu}g/mL$). The polar phase of the extract showed antioxidant activity. The oil showed antimicrobial activity against Streptococcus pneumoniae, Clostridium perfringens, Candida albicans, Mycobacterium smegmatis, Acinetobacter lwoffii and Candida krusei while the water-insoluble parts of the methanolic extracts exhibited slight or no activity. This study confirms that the essential oil of Ledum palustre L. possesses antioxidant and low antimicrobial properties in vitro.

• 약학회지
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• 제18권2호
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• pp.133-144
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• 1974

The metal indicator, acidic azo dyes NN, EBT and Calcon are utilized to analyse quantitatively chlorpheniramine, tripelenamine and diphenhydramine forming insoluble ion pair in aqueous solution at proper pH values between the acidic azo dyes and the sample molecules, these compexes are extracted by organic polar solvents, and organic layer is determined spectrophotometrically. Generally, the absorption maxima of the complexes are shifted to longer wavelengths compare to the absorption maxima of the dyes themselves. The binding ratio of the ion pair forming complex molecules in chloroform soln, are as follows ; NN-antihistamines (chlorpheniramine, tripelennamine, diphenhydramine) are NN-1 to antihisamine-1, EBT-antihistamines are EBT-2 to antithistamines a and Calcon-antihistamines are Calcon-3 to antithistamines-1. These coomplexes in chloroform soln. are very stable, and show higher absorbance than the other organic polar solvents. The binding state of complexes were presumed intermolecular hydrogen bond by their infrared spectra. In the mixture solution of three samples, the aqueous phase is buffered at pH 1.0, and benzene is used to extract ion pair of diphenhydramine EBT complex selectively. At pH 1.0 of aqueous layer, Calon-diphenhydramine complex is also extracted selectively by benzene. However, in this case very small amount of chlorpheniramine-calcon calcon simultaneously. The binding state of diphenhydramine-EBT and diphenhydramine-calcon in benzene are smae as the complexes in chloroform. But the absorption maxima of the complexes in benzene are shifted to shorter wavelenlgths than the complexes in chloroform.

• 한국약용작물학회지
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• 제17권6호
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• pp.434-438
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• 2009

For the development of a new antidementia functional food or alternative drug using agricultural products, Job's tears (Coix lachrymajobi L.), which shows high acetylcholinesterase (AChE) inhibitory activity (55.1%) was selected and the extraction conditions of AChE inhibitor were optimized. AChE inhibitor of Job's tears was maximally extracted when it was treated with 60% methanol at $40^{\circ}C$ for 6 h. The AChE inhibitor of the methanol extracts was partially purified by systematic solvent extraction, thin layer chromatography, silica gel chromatography and reverse-phase HPLC and the partial purified AChE inhibitor with inhibitory activity ($IC_{50}$) of $0.608\;{\mu}g$ was obtained. The partial purified AChE inhibitor was soluble in methanol and hexane, and insoluble in water. Its maximum absorption spectra was 230 nm and also it was stable in the range of $30^{\circ}C$ and $70^{\circ}C$ and pH 4.0-8.0 for 1 h.

• Bulletin of the Korean Chemical Society
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• 제18권3호
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• pp.318-323
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• 1997

ο-(2-Vinyloxyethoxy)benzylidenemalononitrile (3a) and methyl ο-(2-vinyloxyethoxy)-benzylidenecyanoacetate (3b), m-(2-vinyloxyethoxy)benzylidenemalononitrile (4a), and methyl m-(2-vinyloxyethoxy)benzylidenecyanoacetate (4b) were prepared by the condensation of ο-(2-vinyloxyethoxy)benzaldehyde (1) and m-(2-vinyloxyethoxy)benzaldehyde (2) with malononitrile or methyl cyanoacetate, respectively. Bifunctional vinyl ether monomers 3a and 3b polymerized quantitatively with radical initiators in γ-butyrolactone solution at 65 ℃, while meta-isomers 4a and 4b gave lower yields of polymers under the same conditions. The polymers 5-6 obtained from the monomers 3-4 were insoluble in common solvents due to cross-linking. Under the same polymerization conditions ethyl vinyl ether polymerized well with model compounds ο-methoxybenzylidenemalononitrile 7a, methyl ο-methoxybenzylidenecyanoacetate 7b, m-methoxybenzylidenemalononitrile 8a, and methyl m-methoxybenzylidenecyanoacetate 8b, respectively, to give 1:1 alternating copolymers 9-10 in high yields. Cross-linked polymers 5-6 showed a thermal stability up to 300 ℃, and showed a double phase degradation pattern in their TGA thermograms. Polymers 5-6 showed broad endothermic bands around 75-110 ℃ without any characteristic Tg peaks in DSC thermograms. Alternating copolymers 9-10, except copolymer 9b were soluble in common organic solvents. The inherent viscosities of polymer 9-10 were in the range of 0.35-0.62 dL/g. Polymer films cast from acetone solution were cloudy and tough and Tg values obtained from DSC thermograms were in the range of 118-165 ℃.

• 농업과학연구
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• 제39권3호
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• pp.387-393
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• 2012

In the present study, ${\beta}$-glucanase-assisted extraction of starch from glutinous barley(Hinchal ssalbory) was investigated. ${\beta}$-glucanase was added to a coarse starch suspension obtained after wet milling in the starch extraction process. It was found that in the isolated starch with enzyme treatment, protein content was lower by 0.03%, compared to that with non-enzyme treatment. More importantly it was observed that the extraction yield of starch from enzyme treatment was found to be about 12% higher than the one from non-enzyme treatment (enzyme treated: 90.56%, non-enzyme treated: 78.46%). In order to elucidate this finding, the mass-balance determination of starch in each extraction step was carried out and found that the enzyme treatment might influence on the insoluble residues(R3 and R4 fractions) to hydrolyze ${\beta}$-glucan and other materials (e.g., mucilages etc.), thereby facilitated the separation of starch from it and a next filtration process. With a phase-contrast microscope it was observed that the isolated starch with enzyme treatment contained small starch granules more than the one with non-enzyme treatment and this might result in higher extraction yield observed with the former. In order to confirm this hypothesis, further experiments would be necessary.

• 한국지하수토양환경학회지:지하수토양환경
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• 제16권5호
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• pp.42-52
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• 2011

In this laboratory study, the changes in gas-exposed perchloroethene (PCE) saturation in sand during a PCE removal process were measured using gaseous tracers. The flux of fresh air through a glass column packed with PCEcontaminated, partially water-saturated sand drove the removal of PCE from the column. During the removal of PCE, methane, n-pentane, difluoromethane and chloroform were used as the non-reactive, PCE-partitioning, water-partitioning, and PCE and water-partitioning tracers, respectively. N-pentane was used to detect the PCE fraction exposed to the mobile gas. At water saturation of 0.11, only 65% of the PCE was found to be exposed to the mobile gas prior to the removal of PCE, as calculated from the n-pentane retardation factor. More PCE than that detected by n-pentane was depleted from the column due to volatilization through the aqueous phase. However, the ratio of gas-exposed to total PCE decreased on the removal of PCE, implying gas-exposed PCE was preferentially removed by vaporization. These results suggest that the water-insoluble, PCE-partitioning tracer (n-pentane in this study), along with other tracers, can be used to investigate the changes in fluid (including nonaqueous phase liquid) saturation and the removal mechanism during the remediation process.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2005년도 2005 Annual Meeting & International Symposium
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• pp.230-232
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• 2005

The formation of insoluble aggregation of the recombinant kringle fragment of human apolipoprotein(a), rhLK8, in endoplasmic reticulum was identified as the rate-limiting step in the rhLK8 secretion in Saccharomyces cerevisiae. To analyze the protein secretion pathway, some of yeast genes closely related to protein secretion was rationally selected and their oligomer DNA were arrayed on the chip. The expression profiling of these genes during the induction of rhLK8 in fermentor fed-batch cultures revealed that several foldases including pdi1 gene were up-regulated in the early induction phase, whereas protein transport-related genes were up-regulated in the late induction phase. The coexpression of pdi1 gene increased rhLK8-folding capacity. Hence, the secretion efficiency of rhLK8 in the strain overexpressing pdi1 gene increased by 2-fold comparing in its parental strain. The oligomer DNA chip arrayed with minimum number of the genes selected in this study could be generally applicable to the monitoring system for the heterologous protein secretion and expression in Saccharomyces cerevisiae. With the optimization of fed-batch culture conditions and the alteration of genetic background of host, we obtained extracellular rhLK8 at higher yields than with Pichia pastoris systems, which was a 25-fold increased secretion level of rhLK8 compared to the secretion level at the initiation of this study.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.369-375
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• 2001

Urushiol, a natural monomeric oil, was used to prepare a detergentless micro-emulsion with water and 2-propanol The formation of micro-emulsion was verified by conductivity measurements and dynamic light scattering. The conductivity data showed phase change dynamics, a characteristics of micro-emulsions, and subsequent dynamic light scattering study further confirmed the phenomenon. Average water droplet diameter was 10 nm to 500 nm when the molar ratio of 2-propanol ranged from 0.40 to 0.44 . Earlier studies were performed on toluene and hexane, in which the insoluble substrate in water phase was added to the solvents to be reacted on by enzymes. However, in the present urushiol system, urushiol was used as both solvent and substrate in the laccase polymerization of urushiol. The laccase activity in the system was examined using polymerization of urushiol. The laccase activity in the system was examined using syringaldezine as a substrate, and the activity increased rapidly near the molar ratio of 2-propanol at 0.4, where micro-emulsion started. The activity rose until 0.46 and fell dramatically thereafter. The study of laccase activity in differing mole fractions of 2-propanol showed the existence of an ‘optimal zone’, where the activity of laccase was significantly higher. In order to analyze urushiol polymerization by laccase, a bubble column reactor using a detergentless micro-emulsion system was constructed. Comparative study using other organic solvents systems were conducted and the 2-propanol system was shown to yield the highest polymerization level. The study of laccase activity at a differing mole fraction of 2-propanol showed the existence of an ‘optimal zone’ where the activity was significantly higher. Also, 3,000 cP viscosity was achieved in actual urushi processing, using only 1/100 level of laccase present in urushi.