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문석희;이혜경;진소형;김태공;서재현 인제대학교 2012 仁濟論叢 Vol.27 No.1
SLIS(Software License Integrated Solution) is composed of the 6 parts, Server Agent, Web Server, Client Agent, Client Manager, Service, and Install Shield. For the most part of the project is developed based on MFC platform except for the one, Client Manager, which is built on DDK(Driver Development Kit) and ASP.NET platform. Client Agent program installed on a customer's computer is implemented in 14 different classes. Its main function is able to limit and uninstall the purchased software by using teminateProcess. The purchased softwares expiration dates on customer's computer is being monitored by both Server agent and Client software itself. Client Manager hides the process of Client Agent program which shouldn't be killed by manipulating Process List in Kernel through DDK. To do this, Client Manager is internally implanted and registered in Windows Service to be ran as a part of windows system.
So, Jai-Hyun,Do, Hye-Jung,Rhee, In-Koo The Korean Society for Applied Biological Chemistr 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.5
Aspergillus usamii D5, which produces ${\beta}$-glucosidase to hydrolyze isoflavone glycosides of soybean and Astragali Radix into their aglycones efficiently, was isolated from Korean traditional koji, Nuruk. ${\beta}$-Glucosidase produced by the isolated A. usamii D5 was purified to homogeneity through 80% ammonium sulfate precipitation, DEAE Sephadex A-50 column chromatography, Sephacryl S-200 HR column chromatography, and FPLC equipped with Mono Q and Superose columns. The molecular mass of the purified ${\beta}$-glucosidase was estimated to be about 128 kDa. The purified ${\beta}$-glucosidase was stable between 20 and $45^{\circ}C$ but unstable above $45^{\circ}C$. The optimum temperature of the enzyme was $55^{\circ}C$ at pH 4.5. The purified ${\beta}$-glucosidase was stable over a broad pH range from 3.0 to 6.5, but its stability rapidly decreased at pH over 6.5. Maximal activity was observed at pH 4.5 in 100 mM sodium citrate buffer. The enzyme activity was inhibited to 13.2, 17.8, and 5.0% by the addition of $Cu^{2+}$, $Ag^+$, and $Hg^{2+}$ and was reduced to 3.7 and 40.4% by the addition of glucose and maltose, respectively. The purified ${\beta}$-glucosidase showed the highest level of activity to onitrophenyl-${\beta}$-D-glucoside. The purified ${\beta}$-glucosidase efficiently hydrolyzed isoflavone glycosides such as daidzin and genistin extracted from soybean and formononetin and calycosin extracted from Astragali Radix to their aglycone forms.
Jai Hyunk Ryu,Kyoungsun Seo,Hyun Su So,Sheong Chun Lee,Chang-Hyu Bae 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07
Dandelion has been widely used as a folkloric medicine for treatment of diverse diseases. This study was conducted to evaluate the effect of light spectrum using red (660 nm), blue (460 nm), red and blue mixed (Red : Blue = 6 : 4) LED (light emitting diodes) and fluorescent lamp on growth and functional components of dandelion (Taraxacum officinale). When LED was illuminated to T. officinale cv. Goldenboll, seed germination was delayed, and germination rate was the highest in the control (fluorescent light). The growth (plant height, root length and fresh weight) except leaf number was increased under the LED treatments compared with the control, and the growth promotion was the most effective in the red LED illumination. Total polyphenol contents in dandelion irradiated with the red and blue mixed or the red LED were 121.77 mg% or 115.36 mg%, respectively, which were greater than those in dandelion treated with blue LED and fluorescent lamp. Asparagine, proline, serine, threonine, glutamic acid and arginine were the predominant amino acids in dandelion and total amino acid was the highest under the Red LED illumination. The results indicate that application of the red and the mixed LED illumination promote growth and increase functional components during cultivation of dandelion
( Jai Hyun So ),( Yu Mi Lim ),( Sang Jun Kim ),( Hyun Ho Kim ),( In Koo Rhee ) 한국응용생명화학회(구 한국농화학회) 2013 Journal of Applied Biological Chemistry (J. Appl. Vol.56 No.2
Gamma-aminobutyric acid (GABA) aminotransferase (gabT) and succinic semialdehyde dehydrogenase (gabD) genes from Pseudomonas fluorescens KCCM 12537 were cloned into a single pETDuet-1 vector and co-expressed in Escherichia coli BL21(DE3) simultaneously. The mixture of both enzymes, called GABase, is the key enzyme for the enzymatic analysis of GABA. The molecular mass of the GABA aminotransferase and succinic semialdehyde dehydrogenase were determined to be 52.8 and 46.7 kDa following computations performed with the pI/Mw program, respectively. The GABase activity between pH 6.0 and 9.0 for 24 h at 4oC remained over 75%, but under pH 6.0 decreased rapidly. The GABase activity between 25 and 35oC by the treatment at pH 8.6 for 30 min remained over 80%, but over 35oC decreased rapidly. When the activity against GABA was defined as 100%, the purified GABase activity against 5-aminovaleric acid having a similar structure to GABA showed 47.7% and GABase activity against β-alanine, ε-amino-n-caproic acid, L-ornithi , L-lysine, and L-aspartic acid showed between 0.3 to 2.3%. The GABA content was analyzed with this co-expressed GABase, compared with the other GABase which was available commercially. As a result, the content of GABA extracted from brown rice, dark brown rice, and black rice were 26.4±3.5, 40.5±4.7 and 94.7±9.3 μg/g, which were similar data of other GABase in the error ranges.
( So-hyun Chon ),( Min-a Kim ),( Han-saem Lee ),( Jeong-eun Park ),( Yu-mi Lim ),( Eun-jeong Kim ),( Eun-kyung Son ),( Sang-jun Kim ),( Jai-hyun So ) 한국응용생명화학회(구 한국농화학회) 2019 Journal of Applied Biological Chemistry (J. Appl. Vol.62 No.2
The mulberry tree (Morus alba L.) has been traditionally used in Chinese medicine to treat inflammatory diseases. We investigated the effects of bioconversion on different components of the mulberry tree, and determined changes in the physiological activities. Ethyl acetate-soluble fractions of five different segments (fruit, Mori Fructus; leaf, Mori Folium; twig, Mori Ramulus; root, Mori Cortex; and mistletoe, Loranthi Ramulus) of the mulberry tree show enhanced anti-oxidant effects in the 2,2-diphenyl-1-picrylhydrazyl, and 2,2'-azinobis-(3-ethylvenzothiazoline-6-sulfonic acid) assays, and enhanced anti-inflammatory effects of lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in RAW 264.7 macrophages, after being treated with a crude enzyme extract from Aspergillus kawachii, in the following order of activity: Mori Folium>Mori Cortex>Mori Ramulus>Mori Fructus> Loranthi Ramulus. Ethyl acetate- soluble fraction of mulberry leaves (Mori Folium) that underwent bioconversion was most effective, and was devoid of any cytotoxicity. The fraction was also effective against mRNA expression of LPS-induced proinflammatory cytokines, such as inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-α, interleukin-1β, and interleukin-6. In addition, the fraction was effective in LPSinduced phosphorylation of mitogen-activated protein kinases and IKK, and IκB degradation, followed by translocation of the nuclear factor-κB from the cytoplasm to the nucleus. Thus, bioconversion increased the anti-oxidative and anti-inflammatory activities of the mulberry leaf.
Molecular characterization of soybean DnaJ-like protein induced by heat shock stress
Hyun-A So,Eunsook Chung,Chang-Woo Cho,Boh-Hyun Yun,Jee-Sook Kang,Yao Ran,Kyoungmi Kim,Jai-Heon Lee 한국작물학회 2007 한국작물학회 학술발표대회 논문집 Vol.2007 No.11
We isolated wound-inducible genes using suppression subtractive hybridization (SSH) method and were able to obtain to clone w123 gene encoding dnaJ like protein. The full-length cDNA of w123 is 689 bp with an open reading frame (ORF) consisting of 163 amino acid (aa). Genomic southern blot confirmed that soybean genome has two copies of w123 gene. Northern blot analysis was also carried out for the gene expression during heat, NaCl, drought, wounding stresses. The expression of w123 gene specifically induced by heat, NaCl, wounding and drought stress. Using GFP fusion vector, w123-smGFP was targeted both to nucleus. For the functional analysis of w123, His-tagged w123 recombinant protein was heterologously expressed in E. coli. The w123 recombinant cells showed enhanced heat tolerance compared to that of vector control cells. We suggest that dnaJ-like w123 protein function as molecular chaperone in the nucleus of the plant cell during various stresses.
Genetic Variability in Proton Beam Induced Mutants of Dandelion (Taraxacum officinale)
Jai Hyunk Ryu,Hyun Su So,Chang-Hyu Bae 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07
Ionizing radiations have been effective mutagen to overcome the limitation of the useful genetic resources in natural environment. This study was conducted to investigate an effect of proton beam on germination, growth patterns in the irradiated dandelion (Taraxacum officinale), and genetic variation in 12 morphological mutants induced in proton-beam irradiated dandelion. Percentage germination rate was drastically decreased over 250Gy. The lethal dose 50 (LD50) of germination was estimated between 250 Gy to 500 Gy. Significant decreases in growth patterns (plant height, number of leaf and fresh weight) were observed by increase of dose (Gy) of proton beam irradiation. According to the correlation analysis between dosage and growth factors, the orders of compactness of correlation were germination, plant height, fresh weight and number of leaf, respectively. Twelve morphological variants such as, dwarf, color, plastid, growth and leaf shape were screened at 50 to 250 Gy of the beam irradiation. As a result of ISSR analysis of the 12 variants, out of 33 bands detected overall, 8 bands were identified to be polymorphic with a rate of 24.2% at the control group. While 33 bands detected overall, 21 bands were identified to be polymorphic with a rate of 63.6% at the proton beam irradiation. The result indicates that the dandelion with proton beam treatment might be promoted variation at DNA level
So, Jai-Hyun,Rhee, In-Koo The Korean Society for Applied Biological Chemistr 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.3
The molecular cloning of the exo-$\beta$-(1,3)-glucanase gene from Pichia guilliermondii K123-1 was achieved by polymerase chain reaction amplification using oligonucleotides designed according to the N-terminal amino acid sequence of purified exo-$\beta$-(1,3)-glucanase and the conserved regions in exo-$\beta$-(1,3)-glucanase from different yeast species. This gene predicts an open reading frame that has no intron and encodes a primary translation product of 408 amino acids. This preproprotein processes a mature protein of 389 amino acids by signal peptidase and a Kex2-like endoprotease. The mature protein shares 54% to 68% amino acid identity with other yeast exo-$\beta$-(1,3)-glucanases of the glycosyl hydrolase family 5. The eight invariant amino acid residues of the active site and signature pattern (IGIEALNEPL) which existed in all Family 5 members were shown in the mature protein of exo-$\beta$-(1,3)-glucanase but the fifth amino acid (LIVMGST) in the Family 5 signature pattern was changed to A. The cloned exo-$\beta$-(1,3)-glucanase gene was successfully overexpressed in Pichia pastoris X-33 and purified by Ni-NTA His-bind resin chromatography. The molecular mass of the overexpressed enzyme was determined to be approximately 44 kDa. The optimum pH and temperature for activity was 4.5 and $45^{\circ}C$, respectively. This enzyme showed the highest activity toward laminarin (apparent Km, 5.24 mg/mL; Vmax, $7.75\;U/{\mu}g$ protein) among the physiological substrates and 4-methylumbelliferyl-$\beta$-D-glucoside (apparent Km, 8.67 mM; Vmax, $8.99\;U/{\mu}g$ protein) among the chromogenic substrates.