• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.33-43
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• 2013

A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

• Reproductive and Developmental Biology
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• v.35 no.2
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• pp.137-141
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• 2011

These study was to investigate the in vitro fertilization and viability of fresh and vitrified oocytes. Also, the developmental capacity of IVF and intracytoplasmic sperm injection (ICSI) oocytes were investigated. Then vitrification was performed with the use of 20% ethylene glycol + 20% DMSO + 0.5 M sucrose + 10% FCS + TCM-199 medium. Vitrification immature oocytes are cultured in vitrification solution for 10 min afterwards transferred to expose at room temperature for 5 min. and transferred to the ice water for 5 min. The oocytes were sealed in a 1.0 mm straw and placed in a $LN_2$ container. Frozen oocytes were rapidly thawed in a water bath at $30{\sim}35^{\circ}C$, and then placed in TCM-199 medium containing 0.5 M sucrose for 5 min each, respectively, at $38^{\circ}C$. After being washed for 2~3 times, using fresh medium the oocytes were cultured in TCM-l99 medium supplemented with 5% FCS at $38^{\circ}C$ in 5% $CO_2$ and air. The normal morphology of fresh and vitrified-thawed oocytes were $87.1{\pm}2.1%$ and $54.8{\pm}2.5%$, respectively. The viability rates of fresh and vitrified-thawed oocytes were $70.0{\pm}2.2%$ and $41.9{\pm}2.6%$, respectively. Viability rates of vitrified-thawed oocytes were lower than that of fresh follicular oocytes (p<0.05). The in vitro maturation rates of fresh and vitrified oocytes were $45.1{\pm}3.6%$ and $28.9{\pm}4.4%$, respectively. The IVF rates of fresh follicular and vitrified-thawed oocytes were 34.00.2% and $20.2{\pm}2.6%$, respectively. The in vitro maturation and fertilization rates of vitrified-thawed oocytes were lower than those of the fresh follicular oocytes (p<0.05). A total of 350 oocytes were fixed and stained after co-incubation with spermatozoa, of which 88 had identifiable nuclear material. After IVF for 20 hrs, $25.1{\pm}3.4%$ of the oocytes found to have been penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, and 105 oocytes contained identifiable nuclear material. After IVF and ICSI for 20 hrs, $34.3{\pm}3.4%$ and $59.0{\pm}2.0%$ of the oocytes were found to have been penetrated by spermatozoas. The developmental rates upon ICSI were significantly higher than those of the IVF method (p<0.05).

• Biomolecules & Therapeutics
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• v.28 no.4
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• pp.354-360
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• 2020

The hair cycle (anagen, catagen, and telogen) is regulated by the interaction between mesenchymal cells and epithelial cells in the hair follicles. The proliferation of dermal papilla cells (DPCs), mesenchymal-derived fibroblasts, has emerged as a target for the regulation of the hair cycle. Here, we show that vanillic acid, a phenolic acid from wheat bran, promotes the proliferation of DPCs via a PI3K/Akt/Wnt/β-catenin dependent mechanism. Vanillic acid promoted the proliferation of DPCs, accompanied by increased levels of cell-cycle proteins cyclin D1, CDK6, and Cdc2 p34. Vanillic acid also increased the levels of phospho(ser473)-Akt, phospho(ser780)-pRB, and phospho(thr37/46)-4EBP1 in a time-dependent manner. Wortmannin, an inhibitor of the PI3K/Akt pathway, attenuated the vanillic acid-mediated proliferation of DPCs. Vanillic acid-induced progression of the cell-cycle was also suppressed by wortmannin. Moreover, vanillic acid increased the levels of Wnt/β-catenin proteins, such as phospho(ser9)-glycogen synthase kinase-3β, phospho(ser552)-β-catenin, and phospho(ser675)-β-catenin. We found that vanillic acid increased the levels of cyclin D1 and Cox-2, which are target genes of β-catenin, and these changes were inhibited by wortmannin. To investigate whether vanillic acid affects the downregulation of β-catenin by dihydrotestosterone (DHT), implicated in the development of androgenetic alopecia, DPCs were stimulated with DHT in the presence and absence of vanillic acid for 24 h. Western blotting and confocal microscopy analyses showed that the decreased level of β-catenin after the incubation with DHT was reversed by vanillic acid. These results suggest that vanillic acid could stimulate anagen and alleviate hair loss by activating the PI3K/Akt and Wnt/β-catenin pathways in DPCs.

• Korean Journal of Food Science and Technology
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• v.21 no.6
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• pp.808-814
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• 1989

To produce ${\gamma}-linolenic$ acid by a mold, cultural conditions of Mortierella isabellina IFO 8183 were investigated. It was found that the increase of initial pH resulted in the decrease of the ${\gamma}-Linolenic$ acid content and the increase of the C/N ratio of medium resulted in the increase of the lipid content. Addition of sodium acetate into the medium resulted in the increased of cell yield, lipid yield, ${\gamma}-Linolenic$ acid content and ${\gamma}-Linolenic$ acid productivity. Under the optimum coditions(glucose, $NH_4NO_3$, C/N ratio of 40, pH 6.0, $30^{\circ}C$ and 0.5% of sodium acetate), the following results were obtained: cell yield, 0.347(g dry biomass/g glucose; lipid yield, 0.18(g lipid/g glucose); lipid content, 0.52(g lipid/g dry biomass); ${\gamma}-linolenic$ acid content, 60(mg ${\gamma}-linolenic$ acid/g lipid); maximum ${\gamma}-linolenic$ acid concentration, 347mg/l after incubation of 8 days.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.136-141
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• 2005

A bench-scale feasibility study was conducted with solid farming feed to evaluate a treatment process for microbiological removal of nitrogen (N) and phosphorus (P). Strains, Bacillus sp. CK-10 and Bacillus sp. CK-13, were originally isolated from water samples of shrimp farming pond. Simultaneous removal of N/P in marine media was monitored in the co-cultures, CK-10 and CK-13. As the results, $400\;{\mu}M\;NH^{+}_4$ and $400\;{\mu}M\;NO^{-}_2$ were eliminated within 12 hours and $NO^{-}_3$ within 36 hours, and $500\;{\mu}M\;PO^{-3}_4$ was completely disappeared within 36 hours from the media. Cultures of CK-10 and CK-13 were applied for removal of N/P leached from shrimp farming fred. HPAEC-PAD system was used to analyze sugars in farming feed, resulting in resolution of various sugars including glucose, galactose, galatosamine, mannose, and fucose. $0.2\%$ (w/v) Pulp densities of the farming feed contained approximately $33.3\;{\mu}M\;NH^{+}_4,\;12.9\;{\mu}M\;NO^{-}_2.\;81.5\;{\mu}M\;NO^{-}_3\;and\;248\;{\mu}M\;PO^{-3}_4$ which could dissolved within 72 hours of leaching in aqueous solution followed by bacterial removal. Complete bacterial removal of N/P was achieved within 84 hours at $0.2\%$ of the feed in co-cultures, whereas single cultures removed to incompletion of N/P during the incubation period. This work demonstrated that test cultures, CK-10 and CK-13 showed effective removal of N/P derived from shrimp farming feed.

• Korean Journal of Food Science and Technology
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• v.47 no.4
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• pp.525-533
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• 2015

The effect of the hot water extract of defatted green tea seed cake (GTSE) on lipid metabolism and the underlying mechanisms of lipolysis in mature 3T3-L1 adipocytes were investigated. In this study, we found that the naringenin content of GTSE was 5.5 mg/g; however, catechins were not detected. The intracellular lipid droplets were stained with Oil Red O dye and quantified. Compared to the control, lipid accumulation was significantly decreased by 52%, and intracellular triglyceride (TG) level was reduced by 33% after treatment with GTSE at a concentration of $40{\mu}g/mL$. To determine the mechanism of reduction in TG content, we determined the level of fatty acid synthase (FAS), phosphorylation of adenosine monophosphate-activated protein kinase (AMPK), and acetyl-coenzyme A carboxylase (ACC) in the cell model. Incubation of the 3T3-L1 adipocytes with GTSE stimulated AMPK and ACC phosphorylation in a dose-dependent manner, and decreased the expression of FAS.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.195-199
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• 1998

AH alkalophilic Bacillus SP. AIk-7, isolated from soil, produced ethylene. The characteristics of this microorganism is the ability to grow well under the alkaline condition, at pH 10.3. This strain is similar to Bacillus alkalophilus in terms of morphological, physiological and biological characteristics. In observation of relationship of cell growth and ethylene production according to incubation times, the ethylene synthesis mostly occur from the late exponential phase to the death phase of growth. The purpose of this paper is to study the effects of various substrates on the biosynthesis of ethylene in the intact cell and the cell-free system by the Bacillus sp. AIk-7. In both intact cell and cell-free extract, optimum conditions for ethylene production was achieved at pH 10.3 and 3$0^{\circ}C$. Ethylene was effectively produced from L-Met and 1-aminocyclopropane-1-carboxylic acid (ACC). In this case, ACC as the substrate on ethylene production were two fold higher than L-met at each concentration of substrates. On the other hand, the cell-free ethylene-forming system was used as a tool for the elucidation of the biochemical reaction involved in the formation of ethylene by Bacillus sp. AIk-7. Ethylene production in the cell-free system required the presence of manganese and cobalt ion to be stimulated a little. The result obtained in this work suggests that L-met and ACC may be a precursor more directly related to bacterial ethylene production than any other substrates tested.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.9
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• pp.851-856
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• 2010

A sulfur utilizing nitrite denitrification process could be placed after the shortcut biological nitrogen removal (SBNR) process. In this study, removal of nitrite using sulfur oxidizing denitrifier was characterized in batch tests with granular elemental sulfur as an electron donor and nitrite as an electro acceptor. At sufficient alkalinity, initial nitrite nitrogen concentration of 100 mg/L was almost completely reduced in the batch reactor within a incubation time of 22 h. Sulfate production with nitrite was 4.8 g ${SO_4}^{2-}/g$ ${NO_2}^-$-N, while with nitrate 13.5 g ${SO_4}^{2-}/g$ ${NO_3}^-$-N. Under the conditions of low alkalinity, nitrite removal was over 95% but 15 h of a lag phase was shown. For nitrate with low alkalinity, no denitrification occurred. Sulfate production was 2.6 g ${SO_4}^{2-}/g$ ${NO_2}^-$-N and alkalinity consumption was 1.2 g $CaCO_3/g$ ${NO_2}^-$. The concentration range of organics used in this experiment did not inhibit autotrophic denitrification at both low and high alkalinity. This kind of method may solve the problems of autotrophic nitrate denitrification, i.e. high sulfate production and alkalinity deficiency, to some extent.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.11
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• pp.1612-1617
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• 2009

Aspergillus sp. 101 was isolated from the Korean traditional soybean paste for the production of a salt-tolerant protease. The optimal condition for the production of a salt-tolerant protease was determined with various energy sources such as carbon, nitrogen, and protein, and at different culture conditions such as temperature, pH, incubation time and NaCl concentration. The most favorable organic nitrogen sources were 2% defatted soybean flour (DSF) and soy protein isolate (SPI). Optimal pH and temperature were pH 6.0 and $25{\sim}27^{\circ}C$, respectively. Therefore, Aspergillus sp. 101 protease was a mild acid (or neutral) protease. Protease production was the highest at 0.1% concentration of $CaCO_3,\;K_2HPO_4$ and Arabicgum. Aspergillus sp. 101 could grow in culture medium at 15% NaCl concentration and produce a salt-tolerant protease even at 7% NaCl. The cell mass and protease activity of Aspergillus sp. 101 cultured in a modified medium was comparatively higher in Czapek dox and protease producing media. Hence, Aspergillus sp. 101 protease can be utilized in soy or fish sauce industry as a salt-tolerant protease starter.

• Korean Journal of Environmental Biology
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• v.31 no.4
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• pp.471-477
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• 2013

Biochar amendment to agricultural soil is regarded as a promising option to mitigate climate change and enhance soil quality. It could sequester more carbon within the soil system and increase plant yield by changing soil physicochemical characteristics. However, sustainable use of biochar requires comprehensive environmental assessment. In this sense, it is important to measure additional greenhouse gas emission from soils after biochar addition. We investigated emissions of $CO_2$, $N_2O$, and $CH_4$ from incubated soils collected from rice paddy and cultivated grassland after amendment of 3% biochar (wt.) produced from rice chaff. During incubation, soils were exposed to three wet-dry cycles ranging from 5~85% soil gravimetric water content (WC) to investigate the changes in effect of biochar when influenced by different water levels. The $CO_2$ emission was reduced in biochar treatment compared to the control at WC of 30~70% both in rice paddy and grassland soils. This indicates that biochar could function as a stabilizer for soil organic carbon and it can be effective in carbon sequestration. The $N_2O$ emission was also reduced from the grassland soil treated with biochar when WC was greater than 30% because the biochar treated soils had lower denitrification due to better aeration. In the rice paddy soil, biochar addition resulted in decrease in $N_2O$ emission when WC was greater than 70%, while an increase was noted when WC was between 30~70%. This increase might be related to the fact that available nutrients on biochar surface stimulated existing nitrifying bacterial community, resulting in higher $N_2O$ emission. Overall results imply that biochar amendment to agricultural soil can stabilize soil carbon from fast decomposition although attention should be paid to additional $N_2O$ emission when biochar addition is combined with the application of nitrogen fertilizer.