• Korean Journal of Microbiology
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• v.44 no.2
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• pp.122-129
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• 2008

The purpose of this work was to investigate the synergically bactericidal effects and cellular responses of green tea polyphenols (TPPs) and nalidixic acid (NA) on nalidixic acid-resistant (NAR) Salmonella typhimurium. The bactericidal activities of $>3,500{\mu}g/ml$ TPPs and $<256{\mu}g/ml$ NA were investigated for S. typhimurium of which initial cell number was approximately adjusted to 107 cell/ml. Complete elimination of NAS-S. typhimurium was achieved within 6 hr of incubation at the concentrations of $3,500{\mu}g/ml$ TPP or $256{\mu}g/ml$ NA, whereas only partial bactericidal effect was achieved under the same conditions. However, the combinations of $3,000{\mu}g/ml$ TPPs and $32{\mu}g/ml$ NA against NAS-S. typhimurium and $3,500{\mu}g/ml$ TPPs and $64{\mu}g/ml$ nalidixic acid against NAR-S. typhimurium showed complete removal within 5 hr of incubation. The stress shock proteins (SSPs) were induced at different concentrations of TPP o rNA used as stressors against cell culture of S. typhimurium. The proteins were identified as 70-kDa DnaK and 60-kDa GroEL by SDS-PAGE and Western blot. SSPs induced by the stressors were found to increase in proportion to the TPPs or NA. Scanning electron microscopy analyses revealed the presence of perforations and irregular rod shape with wrinkled surfaces for cells treated with TPPs or NA.

• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.169-177
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• 2005

This study focuses on investigating roles of microorganisms in decontamination of reed rhizosphere in Sunchon Bay, Korea, which is considered one of the marsh and mud environment severely affected by human activities such as agriculture and fisheries. In general, the bay is known to play the role of the buffering zone to reduce the sudden impact or change by environmental stresses. In our initial efforts to elucidate the microbial functions in decontamination process in reed rhizosphere, pure bacteria capable of degrading aromatic hydrocarbons were isolated from reed (Phragmites communis) rhizosphere of Sunchon bay by enrichment culture using either benzene, toluene, ethylbenzene, or xylene (BTEX) as a sole source of carbon and energy. Measurement of the rates of BTEX degradation and cell growth during the incubation in BTEX media under several temperature conditions demonstrated maximized degradation of BTEX at $37^{\circ}C$ in both strains. Both strains were also resistant to all the heavy metals and antibiotics tested in this study, as well as they grew well at $42^{\circ}C$. Identification of the isolates based on 16S rRNA gene sequences, and a variety of phenotypic and morphologic properties revealed that the two strains capable of BTEX catabolism were among Microbacterium sp., and Rhodococcus sp. with over $95{\%}$ confidence, designated Microbacterium sp. EMB-1 and Rhodococcus sp. EMB-2, respectively This result suggested that in the rhizosphere of reed, one of major salt marsh plants they might play an important roles in decontamination process of reed rhizosphere contaminated with petroleum such as BTEX.

• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.147-155
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• 1991

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

• Journal of Korea Foresty Energy
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• v.23 no.2
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• pp.21-28
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• 2004

Three different strains of Lentinula edodes, Sanlim 5-Ho, Sanlim 6-Ho and Nongki 3-Ho, were cultured in the sawdust media of Mongolian oak(Quercu mongolica Fisch) for 90 days under dark and light conditions(each 30 days) and fruiting period(30 days). Weight loss of sawdust media was determined after fungal cultures and the contents of ergosterol in fungal mycelia were quantified by HPLC analysis followed by solvent extraction. Compared with the two other fungal strains$(8\%)$, weight loss of Sanlim 5-Ho was slightly lowered to $7\%$. The level of ergosterol content, a parameter for fungal growth, was continuously enhanced in Sanlim 5-Ho for dark and light incubation periods. However, Sanlim 6-Ho and Nongki 3-Ho recorded the maximized fungal growth under light condition. In fruiting periods the ergosterol contents were lowered in the three strains. Intra- and extracellular enzymes during cultural and fruiting periods were also characterized. The activity of Mn-peroxidase and laccase, which are characteristics enzymes for white rot fungi as lignin degrading enzymes, were determined as a high level overall the periods. As cellulose degrading indicators, the activity of CMCase, avicelase, xylanase and glucanase were detectable in initial incubation period.

• Journal of Life Science
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• v.16 no.4
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• pp.676-682
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• 2006

To develop high efficiency biofertilizer solubilizing insoluble phosphates, lactate dehydrogenase (ldh) gene was isolated from Staphylococcus sp. LJ2. Genetic constructions were carried out using the pGEM-T-easy vector and pHSG398. Recombinant DNA plasmids containing the ldh gene were transferred to Klebsiella sp. DA71-1 by electroporation. The selected transformant was named as a DA71-1/pLYJ. The insoluble phosphates solubilization activity of DA71-1/pLYJ was higher than that of DA71-1 at various culture conditions. Glucose was the best carbon source for insoluble phosphates solubilization among the used carbon sources. Maximal insoluble phosphates solubilizing was found in sucrose minimal (SM) medium containing 3% glucose. The solubilizing activity of DA71-1/pLYJ against three types of insoluble phosphates, such as tri-calcium phosphate, hydroxyapatite, aluminium phosphate, were quantitatively determined. The optimal temperature and initial pH to solubilize insoluble phosphates in the SM medium was $37^{\circ}C$ and pH 5.0, respectively.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.163-168
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• 2000

To remove phosphate accumulated in the soil and water, Acinetobacter lwoffi PO8 possessing a high ability to accumulate phosphate was isolated from a active sludge. Bacterium was cultured in the liquid medium containing $150\;{\mu}g/mL$ of phosphate at $30^{\circ}C$ in different culture conditions to examine intracellular phosphate uptake. The initial pH in the range of $7.5{\sim}8.5$ was effective on the growth and phosphate uptake of the strain. Glycerol and arabinose used as a carbon sources showed 93 and 91% the phsphate uptake, respectively. Among the nitrogen sources, ammonium salt such as $NH_4NO_3$ and $(NH_4)_2SO_4$ was effectively utilized on the phosphate uptake compared with amino compounds. The rate of phosphate uptake of $NH_4NO_3$, and $(NH_4)_2SO_4$, was 95 and 96%, respectively The growth and Phosphate uptake ability in the strain were significantly promoted when metal ions were added in the medium; $Co^{2+}$, however, was not utilized by the strain. The capacity of phosphate uptake was enhanced to $10{\sim}20%$ when arginine, methionine, or lysine was added. Using $^{32}P$ to examine the uptake Pattern of intracellular phosphate, experiment result showed that polyphosphate was largely found in the fraction of intracellular inorganic phosphate of Acinetobacter lwoffi PO8.

• Applied Biological Chemistry
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• v.51 no.1
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• pp.1-5
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• 2008

Salt accumulation in soils of greenhouse due to the massive application of nitrogen fertilizers causes salt stress on the various crops, a serious problem in domestic agriculture. Since the majority of the salinity is nitrate, the excess nitrate should be removed; therefore, a bacterial strain having high capacity of nitrate uptake and identified as Enterobacter amnigenus GG0461 was isolated from the soils of greenhouse. Optimum conditions for the bacterial growth and nitrate uptake were investigated. GG0461 was able to grow without nitrate; however, nitrate facilitated the growth. The rate of nitrate uptake increased at alkaline pH and both growth and nitrate uptake were maximal at pH 8-9. When the initial pH of culture medium was increased to pH 8 or 9, it was decreased to neutral upon bacterial growth and nitrate uptake. These results imply that the major factor mediating bacterial nitrate uptake is a nitrate/proton antiporter. The fact was supported by the effect of nitrate addition in the absence of nitrate, since the addition of nitrate greatly increased the nitrate uptake and rapidly decreased pH of media.

• Childhood Kidney Diseases
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• v.14 no.1
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• pp.71-78
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• 2010

Purpose : Hydronephrosis is found about 30% of children with urinary tract infection (UTI). It can be caused by various conditions, although most childhood hydronephrosis is congenital. This study was performed to investigate the relationship between febrile UTI and hydronephrosis. Methods : We retrospectively reviewed the medical charts of 183 patients diagnosed as UTI between January 2007 and May 2009 at Korea University Guro Hospital. Inclusion criteria were as followings; 1) fever more than $37.5^{\circ}C$ measured in the axilla, 2) positive urine culture, 3) no history of urinary tract anomaly on antenatal sonography and urinary tract infection. We classified the enrolled children into two groups of patients with hydronephrosis (HN) and those without hydronephrosis (NHN). Results : The 80 patients were HN and 103 patients NHN. Hydronephrosis was found in 58 patients with left kidney, 8 right and 14 both kidneys. Most of hydronephrosis were of low grade. Compared with NHN group, initial renal cortical defects on DMSA scan significantly increased in HN group (HN 37.5%, NHN 16.5%, P<0.05). The incidence of VUR was not different between the two groups (HN 22%, NHN 12.1%). White blood cell counts and C-reactive protein were not different between the two groups. Follow-up DMSA scan (about 6 months later after UTI) showed no difference of renal scarring in both two groups. Conclusion : Our data suggests that hydronephrosis in febrile UTI patients is clinically useful for detecting renal cortical defects, but is not associated with follow-up renal scar.

• Proceedings of the Korean Society of Life Science Conference
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• 2001.06a
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• Korean Journal of Plant Resources
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• v.28 no.2
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• pp.243-252
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• 2015

This study was performed to establish a production system for in situ and ex situ conservation of Elsholtzia minima Nakai, an endemic plant grown in Jeju Island. Moreover, this study aimed to identify root-growth characteristics according to the use of pre-treatment agents and seedling growth effect according to fertilizer application. The mean temperature was similar in greenhouse and vinyl-moist chamber, but air humidity was higher in vinyl-moist chamber than in greenhouse. After stem planting of Elsholtzia minima Nakai, initial root growth was observed after 10 days in greenhouse and after 7 days in vinyl-moist chamber. Root growth rate was more rapid in vinyl-moist chamber. Moreover, survival rate, root growth rate and root number was slightly higher in vinyl-moist chamber than in greenhouse, indicating that vinyl-moist chamber is more effective in plant growth. When pre-treatment agents were used to remove root growth-inhibiting substances, a higher root growth rate of more than 95% was found in pre-treatment groups, excluding the group treated with AgNO₃ at 77.5%. Thus, Elsholtzia minima Nakai is thought to have less root growth inhibitors. In the analysis of nitrogen application rate and Osmocote application by seedling container, a difference was found in survival rate and growth according to application rate and container conditions. When Osmocote, a slow release fertilizer, was applied to the soil surface around each culture container, survival rate and the growth of aerial and root parts were most favorable. Thus, Osmocote fertilizer is thought to be desirable for seedling propagation of Elsholtzia minima Nakai.