• Korean Journal of Microbiology
• /
• v.52 no.3
• /
• pp.278-285
• /
• 2016

The bifunctional PheA protein, having chorismate mutase and prephenate dehydratase (CMPD) activities, is one of the key regulatory enzymes in the aromatic amino acid biosynthesis in Escherichia coli, and is negatively regulated by an end-product, phenyalanine. Therefore, PheA protein has been thought as useful for protein engineering to utilize mass production of essential amino acid phenylalanine. To obtain feedback resistant PheA protein against phenylalanine, we mutated by using random mutagenesis, extensively screened, and obtained $pheA^{FBR}$ gene encoding a feedback resistant PheA protein. The mutant PheA protein contains substitution of Leu to Phe at the position of 118, displaying that higher affinity (about $290{\mu}M$) for prephenate in comparison with that (about $850{\mu}M$) of wild type PheA protein. Kinetic analysis showed that the saturation curve of $PheA^{FBR}$ against phenyalanine is hyperbolic rather than that of $PheA^{WT}$, which is sigmoidal, indicating that the L118F mutant enzyme has no cooperative effects in prephenate binding in the presence of phenylalanine. In vitro enzymatic assay showed that the mutant protein exhibited increased activity by above 3.5 folds compared to the wild type enzyme. Moreover, L118F mutant protein appeared insensitive to feedback inhibition with keeping 40% of enzymatic activity even in the presence of 10 mM phenylalanine at which the activity of wild type $PheA^{WT}$ was not observed. The substitution of Leu to Phe in CMPD may induce significant conformational change for this enzyme to acquire feedback resistance to end-product of the pathway by modulating kinetic properties.

• Development and Reproduction
• /
• v.11 no.3
• /
• pp.235-243
• /
• 2007

Human umbilical cord blood(HUCB) contains a rich source of hematopoietic stem cells, mesenchymal stem cells and endothelial cell precursors. Mesenchymal stem cells(MSCs) in HUCB are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. We studied on transdifferentiation-promoting conditions in neural cells and cholinergic neuron induction of HUCB-derived MSCs. Neural differentiation was induced by addingdimethyl sulphoxide(DMSO) and butylated hydroxyanisole(BHA) in Dulbeco's Modified Essential Medium(DMEM) and fetal bovine serum(FBS). Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor(bFGF), retinoic acid(RA) and sonic hedgehog(Shh). MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including $\beta$-tubulin III, GFAP and MBP, was markedly elevated during this acute differentiation. The differentiation rate was about $32.3{\pm}2.9%$ for $\beta$-tubulin III-positive cells, $11.0{\pm}0.9%$ for GFAP, and $9.4{\pm}1.0%$ for Gal-C. HUCB-MSCs treated combinatorially with bFGF, RA and Shh were differentiated into cholinergic neurons. After cholinergic neuronal differentiation, the $\beta$-tubulin III-positive cell population of total cells was $31.3{\pm}3.2%$ and of differentiated neuronal population, $70.0{\pm}7.8%$ was ChAT-positive showing 3 folds higher in cholinergic population than neural induction. Conclusively, HUCB-derived MSCs can be differentiated into neural and cholinergic neurons and these findings suggest that HUCB are alternative cell source of treatment for neurodegenerative diseases such as Alzheimer's disease.

• Korean Journal of Food Science and Technology
• /
• v.17 no.6
• /
• pp.495-499
• /
• 1985

To develope a process by which liquid foods can be stored in the liquid state at the frozen storage temperature, suitable cryoprotectants were selected. Orange juice, chosen as an example of liquid foods, was stored with combined cryoprotectants at $-15^{\circ}C$, and quality changes of orange juice during storage were evaluated. Among 7 cryoprotectants tested, NaCl solution had lower initial freezing point than others, and initial freezing points of glucose, fructose, glycerol, propylene glycol and citric acid were close to each other. Considering flavor quality of orange juice, cryoprotectants suitable for reducing freezing point of orange juice were glucose, fructose, glycerol, and citric acid. Combined cryoprotectants for reducing freezing point of 3 and 4 folds concentrated orange juice to $-15^{\circ}C$ consisted of 10% glucose, 8% frutose, 4.6% glycerol and 3% citric acid, and 5.5% glucose, 4.5% fructose, 4.6% glycerol, and 3% cirtric acid, respectively. When destruction of ascorbic acid, sedimentation volume and sensory flavor score of orange juice stored with combined cryoprotectants at $-15^{\circ}C$ and the control stored at $-18^{\circ}C$ were compared, there were no significant differences. These results indicated that liquid foods with suitable combined cryoprotectants could be stored at $-15^{\circ}C$ or below in the liquid state without adverse effect on quality of the stored products.

• Journal of the Korean Society of Clothing and Textiles
• /
• v.3 no.1
• /
• pp.1-11
• /
• 1979

This is a study on angels' costumes in religious paintings, especially as this relates to the questions of concepts and theological symbolism. Angels, as spiritual creatures in Christian thought, play the role of praising God's glory, as messengers of God, the role of guarding Israel and the Church, and protecting or punishing human beings. Sometimes the angels appear in incarnate form. They display no sexual differences and are not able to procreate. The angels' funtional classification being thus; nevertheless, they are pictured in various costumes and appearances according to characteristics of the paintings. The angel Michael appears as a man of dignity when pictured as a guard; the angel Gabriel in the annunciation is often portrayed as a woman of mystical beauty. Under the Renaissance, the mighty cherubim and seraphim at Yahweh's throne are degraded as plump child-angels, or winged child-heads looking alike Eros or Cupid. They have become playful and all too obviously non-heavenly chrubs, accepted features of the Temple decorations. However, cherubim are often depicted as naked or wrapped around with a piece of cloth and accompanied with wind, which symbolizes the Glory of God. The angels, costumes without seam are hung over or wrapped around the body, and when sewn they are simple and ample enough that they fall in a great many folds. However, by the 14C. angels are mostly dressed in costumes common to all Europe, and after that angels gradually appear in folk costumes; for example Italian, Flemish, etc. Dalmatic, the typical costume of Byzantine often shows up as angels' dresses even after the period. Originally the dalmatic was the Roman tunic to which Eastern influences added. The Roman clavus on the tunic had gradually lost distinction until, by the Imperial epoch, it was worn by the lowest servants. It was proudly therefore, as 'The servants of God', that the early Christians are shown wearing the clavus on their wide, ungirdled, sleeved dalmatics. In addition to their costume, angels have some other distinct charateristics. First, angels have a halo around their head; this symbolizes their holiness. Second, angels wear a narrow diadem or a queen's crown that seems to denote their glorious status close to God's throne. Third, the cloth band across the breast resembles a priest's stole, which suggests the sacred role of a priest and symbolizes the grace santified. Fourth, lilies in the annunciations are symbols of Mary's virginity. chastity, innocence and heavenly bliss. Angels hold palms or olives in their hands. The former denote prosperity. beauty and the Christians' reward after death; the latter represent peace and amity. the imperial crown made of olives means victory. Fifth, angels in paintings always have a pair of wings, which can be traced to scripture where cherubim and seraphim are described as having pairs of wings. Angels' wings often have colors of the rainbow, and the rainbow is compared to God's glory. Sixth, generally artists paint angels' costumes as white, blue, green, gold and purple. Other colors such as red rarely appear. According, to scriptures it is believed that angels should be depicted 'as white as snow'. According to the biblical expressions of angels as lightning, sun or a pillar of fire, angels should be described as creatures of light. Nevertheless being a form of art, religious paintings may differ in their presentation according to an artist's inspiration and intention. Since religious paintings illustrated above were almost all done before the Reformation, symbols of colors used in the Catholic Church will also be mentioned. The white color symbolizes chastity, purity, brightness, delight and divinity. Green represents new birth, eternal life, spiritual revival and the expectance of the grace of God. Blue, the color of sapphires, denotes chastity and truth. Red, the color of rubies, represents divinity, love and religious passion. Violet is the color of dignity, indicating the sovereign, royal or imperial power and the great Sacrifice of Christ. As mentionad above, angels' costumes were expressed in accordance with contemporary patterns or as indicated in the Bible, and accesories and colors correspond with Christian symbols. Therefore these facts should be taken into consideration when it comes to the study of costume history.

• The Journal of the Petrological Society of Korea
• /
• v.8 no.3
• /
• pp.131-148
• /
• 1999

Characteristics and complex structures in the northwest Nevada, U.S.A. are de-veloped due to relative tectonic movement of major tectonostratigraphic terranes. Theresearch area is composed of autochthonous rocks of both Early Triassic Koipato Group and Middle Triassic Star Peak Group, which is located in the Humboldt Range, northwest Nevada, U.S.A. The present research is focused on deformation history, related fabric development, and state of regional paleostress during the Jurassic to Late Cretaceous. The Triassic autochthonous rocks in the Humboldt Range, Nevada, U.S.A. display polyphase deformation due to E- to ESE-directed tectonic transport of the Fencemaker allochthon over autochthonous rocks of the Humboldt Range. Structures involving the Mesozoic foreland deformation are development of intense foliation, different styles of folds, minor thrusts, transposed layering, and strong mylonitization. These tectonic structures are mostly developed along the western flank of the Humboldt Range, and are reported as the first deformation of the Mesozoic foreland in the Humboldt Range, Nevada, U.S.A. Regional principal stress(${\sigma}_1$) is interpreted to be E to ESE between the Jurassic and Early Cretaceous on the basis of orientations of strongly developed $D_1$ structures. The deformation during the Middle to Late Cretaceous, is characterized by development of consistent N- to NNE-trending metamorphic quartz veins, and shear zones parallel to pre-existing $D_1$ foliation. Orientations of metamorphic quartz veins as well as other kinematic indicators are N to NNE and are interpreted as those of regional principal stress(${\sigma}_1$) during the Late Cretaceous. The sense of shear applied in the Humbololt Range is dextral and is caused by reactivation of early-formed $D_1$ structures. These results reflect counterclockwise rotation of regional principal paleostress in the Humboldt Range from the Jurassic to Late cretaceous. Finally, development of both shear band cleavage and S/C mylonitic fabrics indicates that the shear zones in the Humboldt Range reflect involvement of enhanced non-coaxial flow during bulk shortening in mylonitic formation.

• Journal of Plant Biology
• /
• v.39 no.1
• /
• pp.63-69
• /
• 1996

A hairy root clone of Panax ginseng C.A. Meyer, HRB-15 was cultured iu various conditions with 3 L bubble type bioreactor to enhance both growth and ginsenoside production. The hairy roots were more rapidly grown under the dark condition than under the light condition. However, total amount of ginsenoside of hairy roots cultured under the light for 30 days increased 2 folds as compared with the dark condition and was 1.10% based on 6 ginsenosides. Especially, ginsenoside-Re was significantly increased and some ginsenosides except for ginsenoside-Re was slightly reduced. Also, the growth of hairy roots decreased about 30% as compared with the dark condition. In contrast, addition of sodium acetate led to decreased production of ginsenoside and growth of hairy roots under light condition. The influence of potassium dihydrogenphosphate concentration was examined in MS medium and a 1.25 mM concentration was found to be the most appropriate for growth and ginsenoside production under light condition. Two-step process of hairy roots culture with yeast elicitation or without ammonia in culture medium was developed to enhance growth and giusenoside synthesis. $50\;\mu\textrm{g}$ of yeast elicitor per g of fresh weight showed a synergistic effect on the ginsenoside synthesis of hairy roots on 20 days after culture. At that time, the content of total ginsenoside was 1.15%, while the growth of hairy roots decreased 21 % as compared with the dark condition. In addition, when elimination of ammonia on 20 days after culture, the content of total ginsenoside was 1.26% with significant increment of ginsenoside-Rd (0.27%) in addition to ginsenoside-Re and the growth of hairy roots decreased 10% as compared with the dark condition. In this system, we have demonstrated a unique two-step process of hairy root cultures to maximize biomass and secondary metabolites. It has found possibility to enhance ginsenosides production by growing hairy roots in this method.

• The Korean Journal of Food And Nutrition
• /
• v.29 no.3
• /
• pp.438-447
• /
• 2016

This study was conducted in order to investigate the physiochemical properties and antioxidative activity of red ginseng manufactured using the high temperature high pressure (HTHP) process, which is faster and simpler than the conventional process. According to increasing the steaming temperature, pressure and time, the content of minor non-polar ginsenosides, such as Rg3, Rk3, Rh4, Rk1 and Rg5 gradually increased. Also, the contents of acidic polysaccharide, total phenolic compounds and maltol gradually increased. Based on the results of the physiochemical properties and appearance quality, the optimum conditions of HTHP process were estimated as $140^{\circ}C$, $3kg/cm^2$ in 20 min. The total phenolic compounds and maltol contents of the HTHP process red ginseng (1.0% and 2.49 mg%, respectively) were higher than those of conventional red ginseng (0.23% and 0.60 mg%, respectively). In addition, the antioxidative activity was investigated using DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-aziono-bis(3-ethylbenzthiazoline-6-sulfonic acid)) radical scavenging activity. DPPH and ABTS radical scavenging activities of HTHP process red ginseng increased by 3.4 and 3.6 folds, respectively, compared with conventional red ginseng. In addition, total phenolic compounds and maltol contents, as well as the antioxidant activity of the HTHP process red ginseng were similar to black ginseng. The present results suggest that the HTHP process is available for the development of value-added red ginseng products.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
• /
• 2004.05a
• /
• pp.89-119
• /
• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Applied Biological Chemistry
• /
• v.42 no.1
• /
• pp.20-28
• /
• 1999

To improve Bacillus strains producing biopolymer, conditions for protoplast formation and regeneration were investigated in biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. Bacillus subtilis K-1 mutant (SM-2) and Bacillus coagulans mutants (CM-12) were marked auxotrophic and antibiotics-resistant (SM-2) and an antibiotics-resistant mutants, respectively. To formate protoplasts derived from the mutants, conditions were established as follows. For B. subtilis mutant SM-2, its culture in mid-logarithmic phase was added with penicillin G (1.0 unit/ml) and further reacted for 1.5 hr. Cells were collected and then treated in lysis fluid (pH 7.0) containing 0.4 M sucrose and lysozyme $25\;{\mu}g/ml$ for 40 min at $37^{\circ}$. Protoplast formation was very successful (99.6%) and the ratio of cell wall regeneration was 2.4%. For Bacillus coagulans mutant CM-12, its mid-logarithmic phase culture was treated with penicillin G (0.3 unit/ml) and glycine (0.5%) for 1hr. Cells were collected and then resuspended in lysis buffer (pH 7.0) containing 0.6 M lactose and lysozyme $(300\;{\mu}g/ml)$ for 30 min at $37^{\circ}$. Protoplast formation was also successful (90.8%) and cell wall regeneration ratio was similar to SM-2 (2.2%). To improve regeneration frequency, regeneration medium was obtained as followed condition,. Cell wall regeneration was improved 2-4 folds with 5.1% for B. subtilis SM-2 and 10.3% for B. coagulans CM-12 when protoplasts mixed with soft top agar(0.4%) was overlaid onto trypticase soy broth medium containing 0.4 M sucrose, 0.7% casamino acid, 1% PVP, 25 mM $MgCl_2,\;25\;mM\;CaCl_₂$ and 1.5% agar.

• The Korean Journal of Pesticide Science
• /
• v.11 no.2
• /
• pp.125-130
• /
• 2007

The development of resistance to acequinocyl was found in the population of the twospotted spider mite, Tetranychus urticae, collected from rose greenhouses in Gimhae, Gyeongnam province in January 2001. This pest is reared on 5 years treated with acequinocyl (over 200 times), and increased 87.8 folds in resistance as compared to susceptible strain (S). Inheritance of acequinocyl resistant strain (R) and cross resistance of this strain to 8 acaricides against T. urticae adults and eggs was investigated. There were differences of susceptibility in the acequinocyl concentration-mortality relationships in $F_1$ progenies obtained from reciprocal cross with the S and R strain ($S(female){\times}R(male)$, $R(female){\times}S(male)$). Degrees of dominance were -0.75, -0.57 in $F_1$ progenies of adult and egg of $S(female){\times}R(male)$. Inheritance in $F_1$ progenies of $S(female){\times}R(male)$ was incomplete recessive. Degree of dominance were 0.81, 0.45 in $F_1$ progenies of adult and egg of $R(female){\times}S(male)$, respectively. These results suggest that inheritance of acequinocyl resistance is controlled by a complete dominance. The R strain exhibited cross resistance of 1.1 and 0.9 fold to amitraz, bifenazate, and negatively correlated cross resistance of 0.08 fold to emamectin benzoate in adult females. The R strain showed cross resistance of 37.7, 14.0, and 26.2 fold to amitraz, milbemectin and spriodiclofen in eggs, respectively. Particularly it showed high levels of cross-resistance to pyridaben with 6538.3 fold. These chemicals showed negatively correlated cross-resistance exhibited 0.4, 0.3, and 0.2 fold to ahamectin, bifenazate, and emamectin benzoate in eggs.