• Journal of Life Science
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• v.18 no.1
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• pp.136-142
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• 2008

A recombinant Pichia pastoris carrying double expression cassette of Rhodotorula glutinis epoxide hydrolase(RgEH) gene was developed and used for preparing enantiopure (S)-styrene oxide from racemic mixture of styrene oxide. BglII restriction site of original RgEH gene (pPICZ B/RgEH #2) of previous report was mutated using PCR technique for the construction of double expression cassette containing promoter ($P_{AOX1}$), EH gene and transcription terminator ($TT_{AOX1}$) in pPICZ C vector. Double expression cassette with RgEH was inserted into the chromosomal DNA of P. pastoris. $V_{max}$ ($2.2{\mu}mol\;min^{-1}mg\;dcw^{-1}$) on (R)-styrene oxide of P. pastoris with double expression cassette was about 6-fold higher than that ($0.4{\mu}mol\;min^{-1}mg\;dcw^{-1}$) of P. pastoris with single expression cassette. For the determination of the optimal condition, the effects of detergent and temperature on the enantioselective hydrolytic activity and yield of the enantiomer were investigated. When the reaction was performed at $10^{\circ}C$ for 10 min in the presence of 0.5% Tween 20, enantiopure (S)-styrene oxide with 99.9% ee was obtained as the yield 43.4 % from 20 mM racemic sustrate.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.1028-1032
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• 1997

The strain No. 33, which produces cellulose-degrading enzyme, was isolated from soil. Yellow halo was identified when the culture supernatant of the strain was loaded onto agar plate containing 2.0% CMC using paper disc method. From scanning electron microscopic observation, the morphology of the stain was rod-shaped. For identification, several biochemical characteristics were tested, and this strain was identified to Bacillus sp. So, we named this strain as Bacillus sp. No. 33. The maximal growth was observed when the stain was cultured in the medium containing 1.0% glucose, 3.0% yeast extract, 0.5% $KH_2PO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 39 hours with shaking. The maximal enzyme production was accomplished using the medium containing 4.0% CMC, 2.0% yeast extract, 0.5% $KH_2PO_4$, 0.04% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 42 hours with shaking.

• Journal of Applied Biological Chemistry
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• v.59 no.4
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• pp.285-288
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• 2016

Proteolytic enzymes perform hydrolysis of the peptide bonds in the protein and most commonly use in the industry. Pseudoxanthomonas sp. WD12 and WD32 were previously isolated as protease producers from a rotten wood sample. Here, we report the secreted proteolytic enzymes. The optimum enzyme reaction temperature for the secreted crude enzyme from the strain WD12 and WD32 were $50^{\circ}C$ at pH 9.0 and $45^{\circ}C$ at pH 8.0, respectively. The enzyme activities of both strains were increased by addition of KCl, NaCl, $CaCl_2$ or $MnSO_4$, and decreased by addition of $AgNO_3$, $CuSO_4$, $FeCl_3$ or $AlCl_3$. Secreted enzymes of both strains were most strongly inhibited by addition of $FeCl_3$ or $CuSO_4$. Taken together these results, WD12 could be a candidate strain of industrial alkaline protease production.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.390-399
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• 2012

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, $pYES2M{\alpha}$-egIV, and $pYES2M{\alpha}$-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants $IpYES2M{\alpha}$-xegIV was higher than that of transformant IpYES2-xegIV or $IpYES2M{\alpha}$-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that $MF{\alpha}$ signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of $35^{\circ}C$ to $65^{\circ}C$. The optimal reaction condition for EGIV enzyme activity was at the temperature of $55^{\circ}C$, pH of 5.0, 0.75 mM $Ba^{2+}$, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant $IpYES2M{\alpha}$-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.2
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• pp.93-99
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• 2008

Until now, (-)-epigallocatechin-3-gallate(EGCG) is known as the most powerful antioxidant among green tea catechins having many beneficial effects on human skin. Considering that the content of catechins is variable according to many conditions such as solvent, temperature and pressure, we prepared the heat-epimerized-EGCG-mixture (HE-EGCG-mix) containing high content of gallocatechin-3-gallate(GCG) by epimerization during autoclaving process and found out its optimal condition for maximizing conversion from EGCG to GCG. To investigate the effects of EGCG and HE-EGCG-mix on skin barrier function, we performed in vivo experiments with hairless mice. We found that HE-EGCG-mix has more potent stimulating activity than EGCG for the production of involucrin 7(INV7) and for recovery of barrier function in SKH-1 mice. Also, we found that GCG stimulates $PPAR-{\alpha}$ transactivation more effectively than EGCG in vitro by transient transfection assay for $PPAR-{\alpha}$ activation activity. These imply that HE-EGCG-mix consisting of high content of GCG should stimulate more efficiently recovery of skin barrier through PPAR-mediated-kerationocyte differentiation than EGCG. In conclusion, our study may provide a possibility that GCG, the C-2 epimer of EGCG, could be a potentially effective agent for development of new cosmetics or health foods for recovery of skin barrier.

• Korean Journal of Agricultural and Forest Meteorology
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• v.14 no.1
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• pp.28-38
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• 2012

This study was conducted to investigate growth performance, photosynthesis, water use efficiency (WUE), and stomatal conductance ($g_s$) of container seedlings of Liriodendron tulipifera and Zelkova serrata growing under three different irrigation periods (1 time/1 day, 1 time/2 days and 1 time/3 days) for high seedling quality. The root collar diameter and height of L. tulipifera and Z. serrata seedlings were highest with 1 time/1 day irrigation, whereas they were lowest with 1 time/3 days irrigation. The two species showed low drought tolerance. As irrigation period was shortened, biomass and seedling quality index (SQI) of the two species increased. The ratio of height to root collar diameter (H/D) and the ratio of below to aboveground biomass (T/R) of the two species were lower with 1 time/3 days than at other irrigation periods. L. tulipifera and Z. serrata seedlings showed significantly higher photosynthetic capacity with 1 time/1 day irrigation. As irrigation period was shortened, $g_s$ of two species increased, while their WUE decreased significantly (P<0.05) These results show that 1 time/1 day irrigation provides the most optimal water condition for container seedling production of two species and irrigation controlling is very important for growth and quality of container seedlings.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.223-229
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• 2006

To develop multifunctional microbial inoculant, an insluble phosphate-solubilizing bacterium with antifungal activity was isolated from plant rhizospheric soil. On the basis of its morphological, cultural and physiological characteristics and Biolog analysis, this bacterium was identified as Pseudomonas fluorescens RAF15. P. fluorescens RAF15 showed antifungal activities against phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% of glucose, 0.005% of urea, 0.3% $MgCl_2{\cdot}6H_2\;0.01%\;of\;MgSO_4{\cdot}7H_2O\;0.01%,\;of\;CaCl_2{\cdot}2H_2O$, and 0.05% of NaCl along with initial pH 7.0 at $30^{\circ}C$. The soluble phosphate production under optimum condition was 863 mg/L after 5 days of cultivation. The solubilization of insoluble phosphates was associated with a drop in the pH of the culture medium. P. fluorescens RAF15 showed resistance against different environmental stresses like $10-35^{\circ}C$ temperature, 1-4% salt concentration and pH 2-11 range. The strain produced soluble phosphate to the culture broth with the concentrations of 971-1121 mg/L against $CaHPO_4$, 791-908 mg/L against $Ca_3(PO_4){_2}$, and 844 mg/L against hydroxyapatite, respectively. However, the strain produced soluble phosphate to the culture broth with the concentrations of 15 mg/L against $FePO_4$, and 5 mg/L against $AlPO_4$, respectively.

• Journal of Dairy Science and Biotechnology
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• v.29 no.2
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• pp.37-42
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• 2011

The changes in pH, titratable acidity, chromaticity, total count, coliform group and organoleptic properties of the whole market milks (UHT, LTLT) that sold currently on the domestic market were stored after their production at 0, 10, 20, 30 and $40^{\circ}C$ to predict their quality during distribution, and examined prior to the analysis on the correlation of their quality properties and organoleptic preference level and discovery of optimal quality indicator. The investigation of correlation between pH, acidity and preference level of milks depending on respective storage temperature showed significant correlation (p<0.01) for the milk stored at 10, 20, 30 and $40^{\circ}C$, and the higher temperature was directly proportional to the higher correlation coefficient. The correlation between total count and preference level for LTLT milk stored at 0, 10, 20, 30 and $40^{\circ}C$ showed high correlation coefficient at every high temperature condition respectively as R=0.81, R=0.91, R=0.96, R=0.90 & R=0.99, and the correlation coefficients were also significant level for the UHT milk irrespective of their storage temperature except $0^{\circ}C$. Accordingly, the changes in total colonies turned out to be suitable to be the quality indicator for the quality prediction of the milk on the distribution.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.9
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• pp.6142-6151
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• 2015

This study is conducted to develop and optimize fruit spirits by using Schizandra chinensis fruit which are rarely used as spirits in the world. For this purpose, the raw material, main compounds of fermented mash and, quality characteristics of distilled liquor prepared with different distillation methods were analyzed. The studt period lasted six months and the results were follows. Schizandra chinensis was not suitable for alcohol fermentation because the sugar concentration and acidity were low. Therefore, the fermentation condition was appropriate to mix with the weight ratio of fresh Schizandra chinensis fruit, water and sugar as 1:1.5:0.25, and was fermented the mix by adding 0.06 w/w% of $(NH_4)_2HPO_4$ compared to the must total weight. It was also examined to analyze product characteristics of fruit spirits prepared with different methods of distillation(column-pot still, pot still, vacuum still). The result showed that fruit spirits made by vacuum still with fresh Schizandra chinensis fruit indicated the best product quality while the distillation of column-pot still showed the best yield.

• The Korean Journal of Mycology
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• v.38 no.2
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• pp.184-188
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• 2010

The goal of this study was to screen a useful yeast for Korean traditional pear Yakju (KTPY) brewing and develop its brewing process. Cooked non-glutinous rice and nuruk were mixed, and added into pear juice with various Saccharomyces cerevisiae and then fermented at $25^{\circ}C$ for 7 days. Among several alcohol fermentation yeasts, ethanol contents was the highest in pear Yakju made by S. cerevisiae K-7 and also showed high ethanol content in pear Yakju which was made by commercial S. cerevisiae C-2. Therefore, we selected S. cerevisiae K-7 and S. cerevisiae C-2 as suitable yeasts for brewing of KTPY. Maximal ethanol production (10.4%) was obtained when cooked non-glutinous rice (100 g) and nuruk (30 sp/g) were mixed and added into pear juice (600 ml) with S. cerevisiae K-7 (5%) and fermented at $25^{\circ}C$ for 7 days and also its antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity was 57.2%. Addition of antihypertensive starchy materials into the mash was not affected in ACE inhibitory activity and total acceptability of KTPY.