• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.384-389
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• 1992

Bacillus sp. YBL-7 which had been isolated from ginseng root-rot suppressive soil was able to antagonize Fusarium solani causing ginseng root-rot by their antibiotic substance. In order to develop multifunctional antagonist on Bacillus sp. YBL-7 as a biocontrol agent against Fusarium salam', optimal conditions for protoplast transformation system of Bacillus sp.YBL-7 by the vector plasmid pE194 were investigated. The protoplasts of Bacillus sp. YBL-7 were obtained at best efficiency by treatment with 200${\mu}g$/ml of lysozyme in the pH 7.0 of SMM buffer for 90 minutes at $40^{\circ}C$. The cell wall of the protoplast was regenerated on the agar plate containing 1.2% agar and 0.7 M mannitol. Under the best condition for protoplast formation and regeneration, the optimal transformation was achieved with 40% polyethylene glycol (M.W. 4000) treatment for 10minutes. The vector plasmid pE194 showed the best transformation frequency at 5$\mu$g/ml of final concentration. The pE194 was very stable over 80% in the transformants.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.142-149
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• 1988

This experiment was carried out to obtain a hybrid with potent ethanol fermenting ability, by means of protoplast fusion between a thermophilic strain (D-71) of Saccharomyces cerevisiae and an osmotolerant strain (SR-S) of Zygosaccharomyces rouxii. The conditions for formation of protoplasts from both strains and for their fusion and regeneration were studied. Favorable conditions for formation of protoplasts from Saccharomyces cerevisiae D-71 were : treatment of the cells at late-exponential phase with 2-mercaptoethanol (l% v/v) for 10 minutes in the presence of 0.5M sorbitol, then incubation for 60 minutes in the set medium containing Zymolyase-20T (4mg/$m\ell$) ; and from Zygosaccharomyces rouxii SR-S were : treatment of the cells at mid-exponential phase with 2-mercaptoethanol (1% v/v) for 10 minutes in the presence of 0.5M or 1M mannitol, then incubation for 120 minutes in the set medium containing Zymolyase-20T(4mg/$m\ell$). The protoplasts of parental cells were fused in the presence of 20mM CaCl$_2$, 0.5M sorbitol and 40% of polyethyleneglycol (M.W 4000), then fusants obtained were selected as regenerated colonies which embedded and grown in the minimal medium containing 3% of agar. The frequencies of fusant formation were 1.2$\times$10$^{-6}$ to 9.1$\times$10$^{-6}$ for the regenerated protoplast.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.297-302
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• 1988

The protoplasts of Saccharomyces cerevisiae D-71, a thermophilic strain and Zygosaccharomyces rouxii SR-S, an osmotolerant strain were fused, and a fusant (FS-RN 1) was selected, then was characterized for its genetic stability, DNA content, cell capacity, growth rate, tolerance to salts and chemicals, $\beta$-fructofuranosidase level and ethanol fermenting activity. After 6 months of preservation, 5.8% of the fusant clones were segregated to parental types. The DNA content and cell capacity of the fusant were greater than those of the parental strains. Lag period of growth for the fusant was longer than those for the parents. The fusant colonies showed pink-color reaction to triphenyltetrazolium chloride(TTC) test. The fusant appeared to have resistances to NaCl at moderate levels between both parental strains, and resistances to KCI, sodium propionate and cycloheximide similar to either one of the parents. $\beta$-Fructofuranosidase activity of the fusant was 24.2$\times$10$^{-3}$/IU/g(dry wt) for 3 days culture. Ethanol yields ofter 4 days of fermentation by the fusant at 3$0^{\circ}C$ were : 6.0%(v/v) from 40% of glucose and 8.8%(v/v) from 20% of sucrose.

• Journal of Korean Society of Forest Science
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• v.73 no.1
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• pp.33-42
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• 1986

This study was carried out to investigate the optimum conditions for isolation and culture of mesophyll protoplasts from Populus alba ${\times}$ P. glandulosa. The results obtained from the experiments are as follows; 1) The suitable concentration of BAP for shoot multiplication was 0.4 mg/l. 2) High yield and viability of isolated protoplasts were obtained by our high enzyme-short time incubation method. 3) Optimum enzyme concentrations for mesophyll protoplast isolation were Cellulase 2%, Macerozyme 0.8%, Hemicellulase 1.2%, Driselase 2%, and Pectolyase Y-23 0.05%. 4) 0.6M mannitol in enzyme solution was the most effective for protoplast isolation and viability. 5) The most adequate pH level of enzyme solution was pH 5.6. 6) The effect of DTT and MES buffer was significant. 7) For protoplast purification, 0.6M sucrose was the most proper concentration. 8) The adding effect of Dextran T40 in floating solution was important. 9) The mesophyll protoplasts isolated through our high enzyme-short time incubation method revealed successful response to culture condition over 3 weeks of culture.

• Journal of the Korean Geosynthetics Society
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• v.17 no.4
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• pp.169-177
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• 2018

Glass fiber reinforced polyester (GFRP) composites are widely used as structural materials in harsh environment such as underground pipes, tanks and boat hulls, which requires long-term water resistance. Especially, these materials might be damaged due to delamination between gelcoat and composites through an osmotic process when they are immersed in water. In this study, GFRP laminates were prepared by surface treatment of UPE (unsaturated polyester) gelcoat by vacuum infusion process to improve the durability of composite materials used in underground pipes. The composite surface coated with gelcoat was examined for surface defects, cracking, and hardness change characteristics in water-immersion environments (different temperatures of $60^{\circ}C$, $75^{\circ}C$, and $85^{\circ}C$). The penetration depth of cracks was investigated by micro CT imaging according to water immersion temperature. It was confirmed that cracks developed into the composites material at $75^{\circ}C$ and $85^{\circ}C$ causing loss of durability of the materials. The point at which the initial crack initiated was defined as the failure time and the life expectancy at $23^{\circ}C$ was measured using the Arrhenius equation. The results from this study is expected to be applied to reliability evaluation of various industrial fields where gelcoat is applied such as civil engineering, construction, and marine industry.

• Applied Biological Chemistry
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• v.49 no.1
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• pp.15-20
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• 2006

EDTA is known to have bacteriocidal effect on Vibrio vulnificus, pathogen of septicemia by osmotic shock in seafoods. Attempts were made to elucidate the bacteriocidal effect of phytic acid (PA) as a substitute for EDTA against V. vulnificus and its inhibition effect on the septicemia, which induces liver damage of the mice by the pathogen. Viable cells of V. vulnificus with the initial titre of $1.7{\times}10^6$ c.f.u. $ml^{-1}$ decreased by 90.6% after 1 min and 99.6% after 5 min in distilled water. The titre decreased by 65.9% and 94.5% in 2 mM solution of $Mg^{2+}$. In 0.1 mM solution of PA, the rate of decrease in titre was 97.4% after 1 min of incubation and 99.8% after 5 min, compared to 95.7% and 99.8% in 0.1 mM solution of EDTA. The bacteriocidal effect of PA solution at a concentration of 1 mM was marked: the rate of decrease in titre was 99.9% after 1 min. In relation to the bacteriocidal effect, PA was evaluated as a potential therapeutic agent for V. vulnificus septicemia in mouse. When the survival periods of mice were investigated by PA and EDTA treatment after the pathogen injection, the group of mice which infected by a low concentration of the strain survived longer than that inoculated at high concentration; also, the ratio of survival was 1.3 times higher in PA than in EDTA, showing that the fatal rate depended on the inoculation concentration. Although survival periods of mice induced with liver damage by carbon tetrachloride and then inoculated with the strain showed a similar trend, the fatal rate of mice was 2 times faster than those inoculated with only pathogen into normal liver, These results indicate that the infection by V. vulnificus was more fatal to those with liver disease. Also, symptoms of hemorrhage and inflammation on the mice with induced liver damage were reduced in case there was phytic acid treatment at each concentration.

• Journal of Chest Surgery
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• v.41 no.5
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• pp.550-562
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• 2008

Background: We attempted to reproduce a previously reported method that is known to be effective for decellularization, and we sought to find the optimal condition for decellularization by introducing some modifications to this method. Material and Method: Porcine semilunar valves, arterial walls and pericardium were processed for decellularization with using a variety of combinations and concentrations of decellularizing agents under different conditions of temperature, osmolarity and incubation time. The degree of decellularization and the preservation of the extracellular matrix were evaluated by staining with hematoxylin and eosin and with alpha-Gal and DAPI in some of the decellularized tissues. Result: Decellularization was achieved in the specimens that were treated with sodium deoxycholate, sodium dodesyl sulfate, Triton X-100 and sodium dodesyl sulfate with Triton X-100 as single-step methods, and this was also achieved in the specimens that were treated with hypotonic solution ${\rightarrow}$ Triton X-100 ${\rightarrow}$ sodium dodesyl sulfate, sodium deoxycholate ${\rightarrow}$ hypotonic solution ${\rightarrow}$ sodium dodesyl sulfate, and hypotonic solution sodium dodesyl sulfate as multi-step methods. Conclusion: Considering the number and the amount of the chemicals that were used, the incubation time and the degree of damage to the extracellular matrix, a single-step method with sodium dodesyl sulfate and Triton X-100 and a multi-step method with hypotonic solution followed by sodium dodesyl sulfate were both relatively optimal methods for decellularization in this study.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.2
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• pp.195-201
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• 2014

Body fluid has been studied for diverse fields like Ringer's solutions, artificial joint fluids, cell growth culture media because it plays a crucial role in controlling body temperature and acts as a solvent for diverse metabolite processes in the body and delivery media of mineral, energy source, hormone, signal and drug from and to cell via blood or lymphatic vessel by osmotic pressure or active uptake. Stratum corneum containing extracellular lipids and NMF (natural moisturizing factor) absorbs atmospheric water residing outside of cells and utilize it to hydrate inside of their own. This process is related to skin barrier function. In this study, we conducted the cell viability test with Cell Bio Fluid $Sync^{TM}$, which mimicks body fluids including amino acids, peptides, and monosaccharides to strengthen skin barrier, and the clinical skin improvement test with cosmetics containing Cell Bio Fluid $Sync^{TM}$. In the cell viability test, HaCaT cell was treated with PBS for 3 hours, followed by the treatment of a cell culture medium (DMEM) and isotonic solution (PBS) and Cell Bio Fluid $Sync^{TM}$ for 3 hours each. Then, MTT assay and image analysis were conducted. In the clinical skin improvement test, twenty-one healthy women participated. Participants applied cosmetics containing Cell Bio Fluid $Sync^{TM}$ on their face for a week and evaluated the skin hydration, skin roughness, brightness and evenness. All measurements were conducted after they washed off their face and took a rest under the constant temperature ($22{\pm}2^{\circ}C$) and constant humidity conditions ($50{\pm}5%$) for 20 minutes. All the data were analyzed by SPSS (version 21) software program. Results showed that Cell Bio Fluid $Sync^{TM}$ improved both the cell viability and in vivo skin conditions such as skin hydration, roughness, brightness and evenness.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.141-146
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• 1995

Changes of physicochemical properties of citron juice prepared by two different extraction methods, rotary-crushing and belt-pressing method, were investigated during the storage at $5^{\circ}C$ and $-20^{\circ}C$. Temperature drop of citron juice extracted by belt-pressing method was faster than that of citron juice prepared by rotary-crushing method and its freezing point was $0.8{\sim}0.9^{\circ}C$. During the storage, pH of stored citron juice with rotary-crushing method was increased up to 3.5 after 6 months storage while that of citron juice extracted by belt-pressing method was not changed significantly during the same storage time. Acidity of rotary-crushed citron juice was reduced a little more than that of belt-pressed citron juice during the storage. However, changes of soluble solid content were influenced largely by the storage temperature than by the extraction method. Contents of formol nitrogen and vitamin C were reduced remarkably in all of stored citron juice and $92{\sim}82%$ of farmol nitrogen and $72{\sim}43%$ of vitamin C were remained after 6 months of storage. Among the changes of color value, L values were reduced in the whole stored citron juice and a and b value had a different change pattern respectively according to the extraction and storage temperature. Changes in the content of both amino acid and fatty acid compositions was also observed after same storage period. Especially, in the case of change of fatty acid composition, content of linoleic acid and linolenic acid were reduced after 6 months storage, while those of palmitic acid, stearic acid and oleic acid were increased.