• Journal of Microbiology
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• v.34 no.4
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• pp.341-348
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• 1996

The regulation of xylE gene expression was examined by using vector promoter and construction of genetically engineered microorganisms (GEMs) for application in microcosm. When the xylE gene wsa subcloned into pBluscript SK(+) under the control of lac promoter (pTY1) in E. coli, and the expression was induced by IPTG, the enzyme activity of catechol 2, 3-dioxygenase was increased 4.7 times more than that of the crude extracts from transformants harboring pTY1. We suggest that the xylE gene has its own promoter at the upstream portion, because it was able to be expressed even in the absence of IPTG. A recombinant plasmid, pSW3a harboring the xylE gene under the T7 promotor, showed the activity of 14.5 units/mg protein, higher than that of parental strain, E. coli PYT1. The xylE gene in recombinant plasmid pSW3a was used as reporter gene for the application in microcosm ecosystem, since it was used for detection of xylE-positive clones by catechol spray on the agar plates. The pSW3a in E. coli was introduced into Pseudomonas patida to construct GEM strain, and examined for the exxpression and functional stability in microcosms.

• KSBB Journal
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• v.18 no.5
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• pp.347-351
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• 2003

The extraction of glycyrrhizic acid from licorice using supercritical carbon dioxide (SCCO$_2$) was investigated with respect to the effects of extraction parameters such as the kind and amount of modifier, temperature, pressure, and extraction time. The conventional organic solvent extraction was also conducted for a quantitative comparison. The content of glycyrrhizic acid in crude extracts was analyzed by HPLC and the yield of glycyrrhizic acid was computed as a weight percent recovery. The optimal pressure and temperature for SCCO$_2$ extraction were found to be 40 MPa and 80$^{\circ}C$, respectively, when SCCO$_2$ was modified with 70% aqueous ethanol. Under the same pressure and temperature, the highest recovery was attained to be 104.57% in the first 60 min when the concentration of 60%, aqueous ethanol in SCCO$_2$ was 15%.

• KSBB Journal
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• v.7 no.3
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• pp.186-190
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• 1992

The compositions of amino acid in the protein and total fatty acid of Dendrolimus spectabilis were analyzed quantitatively by HPLC and GC, respectively. The contents of crude oil and protein from the extracts were 21.00% and 58.47%, respectively. The amount of free amino acids in the protein was 3.65g/100g, and 1.31g/100g of essential amino acids was contained in the free amino acids. The amount of total amino acids in the protein was 41.20g/100g, and 14.75g/100g of essential amino acids were contained in the total amino acid. The compositions of fatty acid in the oil were $C_{18}$=26.81%, $C_{16}$=19.09%, $C_{18:1}$=18.74%, $C_{18:3}$=15.33%, $C_{16:1}$=7.29%, $C_{20}$=5.21% in order, respectively. 45.88% of unsaturated fatty acids were contained in the oil.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.88-94
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• 2008

Two sugar biosynthetic cassette plasm ids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003-OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ $Na^+$] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12-membered ring aglycon (10-deoxymethynolide, 1) and a 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1491-1499
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• 2010

Rhizosphere microorganisms possessing phytase activity are considered important for rendering phytate-phosphorus (P) available to plants. In the present study, the Citrobacter braakii phytase gene (appA) was overexpressed in rhizobacteria possessing plant growth promoting (PGP) traits, for increasing their potential as bioinoculants. AppA was cloned under the lac promoter in the broadhost-range expression vector pBBR1MCS-2. Transformation of the recombinant construct pCBappA resulted in high constitutive phytase activity in all of the eight rhizobacterial strains belonging to genera Pantoea, Citrobacter, Enterobacter, Pseudomonas (two strains), Rhizobium (two strains), and Ensifer that were studied. Transgenic rhizobacterial strains were found to display varying levels of phytase activity, ranging from 10-folds to 538-folds higher than the corresponding control strains. The transgenic derivative of Pseudomonas fluorescens CHA0, a well-characterized plant growth promoting rhizobacterium, showed the highest expression of phytase (~8 U/mg) activity in crude extracts. Although all transformants showed high phytase activity, rhizobacteria having the ability to secrete organic acid showed significantly higher release of P from Ca-phytate in buffered minimal media. AppA overexpressing rhizobacteria showed increased P content, and dry weight (shoot) or shoot/ root ratio of mung bean (Vigna radiata) plants, to different extents, when grown in semisolid agar (SSA) medium containing Na-phytate or Ca-phytate as the P sources. This is the first report of the overexpression of phytase in rhizobacterial strains and its exploitation for plant growth enhancement.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1726-1736
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• 2013

Biosynthesis of indole-3-acetic acid (IAA) via the indole-3-pyruvic acid pathway involves three kinds of enzymes; aminotransferase encoded by aspC, indole-3-pyruvic acid decarboxylase encoded by ipdC, and indole-3-acetic acid dehydrogenase encoded by iad1. The ipdC from Enterobacter cloacae ATCC 13047, aspC from Escherichia coli, and iad1 from Ustilago maydis were cloned and expressed under the control of the tac and sod promoters in E. coli. According to SDS-PAGE and enzyme activity, IpdC and Iad1 showed good expression under the control of $P_{tac}$, whereas AspC was efficiently expressed by $P_{sod}$ originating from Corynebacterium glutamicum. The activities of IpdC, AspC, and Iad1 from the crude extracts of recombinant E. coli Top 10 were 215.6, 5.7, and 272.1 nmol/min/mg-protein, respectively. The recombinant E. coli $DH5{\alpha}$ expressing IpdC, AspC, and Iad1 produced about 1.1 g/l of IAA and 0.13 g/l of tryptophol (TOL) after 48 h of cultivation in LB medium with 2 g/l tryptophan. To improve IAA production, a tnaA gene mediating indole formation from tryptophan was deleted. As a result, E. coli IAA68 with expression of the three genes produced 1.8 g/l of IAA, which is a 1.6-fold increase compared with wild-type $DH5{\alpha}$ harboring the same plasmids. Moreover, the complete conversion of tryptophan to IAA was achieved by E. coli IAA68. Finally, E. coli IAA68 produced 3.0 g/l of IAA after 24 h cultivation in LB medium supplemented with 4 g/l of tryptophan.

• Korean Journal of Food Science and Technology
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• v.17 no.4
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• pp.240-244
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• 1985

Curde $\beta$-amylase was prepared by frationation of the water extracts from Korean ginseng, Panax Ginseng C.A. Meyer, with 0.2-0.6 saturation of ammonium sulfate. The enzyme showed the typical properties of $\beta$-amylase, producing only maltase from starch. The enzyme preparation also showed no maltase activity. The enzyme was stable at the pH 5-9 and at the temperature below $40^{\circ}C$. The enzyme showed the optimum pH at 5.0 and the optimum temperature at $35^{\circ}C$. Its activities had proportional relations with substrate concentration below 12 mg%, showing Km V slues of 4.76 mg%. The enzyme was inhibited by $Ag^{+}$, $Hg^{++}$, $Cd^{++}$, $Cu^{++}$,$ Al^{3+}$, and $Fe^{3+}$.

• The Korean Journal of Food And Nutrition
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• v.26 no.3
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• pp.375-382
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• 2013

The present study is designed to explore an anti-tumor activity on crude extracts of Oplopanax elatus. Water extractions of Oplopanax elatus were performed at $100^{\circ}C$(OeE-100). OeE-100 doses up to $62.5{\mu}g/m{\ell}$ had no cytotoxicity on the tumor cell lines in vitro. In experimental lung metastasis of colon26-M3.1 carcinoma or B16-BL6 melanoma, the prophylactic intravenous ($4{\sim}100{\mu}g/mouse$) or oral (2 mg/mouse) administration of OeE-100 significantly inhibited tumor metastasis as compared with tumor controls. Peritoneal macrophages stimulated with OeE-100 produced various cytokines such as TNF-${\alpha}$, IL-6 and IL-12. In an analysis of NK-cell activities, i.v. administration of OeE-100 ($10{\sim}100{\mu}g/mouse$) significantly augmented the cytotoxicity to YAC-1 tumor cells. Vaccination of mice with boiling-treated tumor cells (BT-vaccine) in combination with OeE-100 ($100{\mu}g/mouse$) showed higher inhibitions in tumor metastasis when compared with the mice of BT-vaccine treatment. In addition, the splenocytes from OeE-100 admixed BT-vaccine immunized mice secreted a higher concentration of Th1 type cytokine such as IFN-${\gamma}$. These results suggested that the OeE-100 stimulated immune system and was a good candidate adjuvant of anti-tumor immune responses.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.4
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• pp.620-627
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• 2011

We evaluated the hepatoprotective activity of crude hot-water extracts of the traditional Korean medicinal herb, Phellodendri Cortex (PC; Bark of Phellodendron amurense RUPR.), in an experimental model of hepatic damage induced by carbon tetrachloride ($CCl_4$). The serum marker of liver damage, sGOT, sGPT and sALP, were significantly decreased in the liver of the PC treated rats, compared with that of $CCl_4$ treated group. The histological observation of liver section of rats, showed the same protective effect of PC treatment. And the protective activity of PC was more significant in the post-treated group than pre-treated group. The significant decrease of malodialdehyde and increase of glutathion, catalase activity were observed in the liver homogenate of PC treated rats. Based on these findings, it is suggested that PC has potent hepatoprotective effects and the mechanism of the protection may be related to antioxidation pathways.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.30 no.1
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• pp.53-62
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• 1985

A radioimmunoassay technique has been developed for the determination of abscisic acid (ABA) in crude extracts from germinating rice (Oryza sativa L.). By this method, the changes in ABA level of rice during germination was investigated. The ABA content in rice seeds was found to be 76.5ng/g dry weight in Dong-jin variety and 91.1ng/g dry weight in Sam-gang variety. A rapid decrease in ABA content of rice occurred during germination within 24 hours after seed imbition. The decreasing rate of ABA content during germination showed a significant direct proportion to the imbibition temperature and water-absorbing rate of rice. The decrease in ABA content during germination was found to be caused partly by an elution of ABA from the tissue to the imbibing fluid, and partly by a metabolic conversion of ABA to another compounds. The germination process of rice occurred only when the tissue ABA level decreased below a certain level, and the decreasing rate of ABA level during germination correlated with the ability for germination at low temperature of rice.