• Journal of Microbiology and Biotechnology
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• v.8 no.1
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• pp.34-41
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• 1998

A method was developed for the controlled expression of an antimicrobial peptide magainin in Escherichia coli. A series of concatemeric magainin genes was constructed with a gene amplification vector, and fused to the 3'end of malE gene encoding the affinity ligand, E. coli maltose-binding protein (MBP). The construct directed the synthesis of the fusion protein with the magainin polypeptide fused to the C-terminus of MBP. The fusion protein was expressed in a tightly regulatable expression system which was under the control of an invertible promoter. The MBP-fused magainin monomer was expressed efficiently. However, the expression level of the MBP-fused magainin in E. coli decreased with the increasing size of multimers possibly because of the transcription and translation inhibition by the multimeric peptides. After purification using an amylose affinity column, the fusion protein was digested by factor Xa at a specific cleavage site between the monomers. The recombinant magainin had an antimicrobial activity identical to that of synthetic magainin. This experiment shows that a biologically active, antimicrobial peptide magainin can be produced by fusing to MBP, along with a promoter inversion vector system.

• Korean journal of food and cookery science
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• v.14 no.1
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• pp.9-15
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• 1998

The inhibitory effect of clove (Eugenia caryophyllata Thumb) on the growth of Escherichia coli 0157:H7 was determined. Tryptic soy broth (TSB) containing 0∼0.5% (w/v) of clove was inoculated with 10/sup/5∼10/sup/7 CFU/ml of E. coli and incubated at 5 different temperature (35, 5, -20, 50 and $55^{\circ}C$). The growth of E. coli was not inhibited at 0.1% clove and growth occured at 0.3% after a prolonged lag period while viable cells of E. coli decreased at 0.5% clove during storage at $35^{\circ}C$. During 32 days of refrigerated storage at $5^{\circ}C$, survivors of E. coli were decreased with the progress of time and increasing clove concentration. At the presence of 0.3 or 0.4% clove, bacterial cells were dead at the end of refrigerated storage. During frozen storage at -$20^{\circ}C$, survivors of E. coli at the presence of 0∼0.4% clove were decreased 2.9∼4.07 log cycles for 4 days of early period and then decreased 1.0∼2.1 log cycles through the frozen storage. There were small changes in populations of E. coli in TSB between different concentrations of clove during frozen storage. The D-values for E. coli at $50^{\circ}C$ were 105.26, 22.47, 13.76, 11.14 and 10.17 min at clove 0, 0.1, 0.2, 0.3, 0.4%, respectively. The D-values for E. coli at $55^{\circ}C$ were 10.75, 8.95, 7.40, 5.96 and 4.96 min at clove 0, 0.1, 0.2, 0.3, 0.4%, respectively. Antibacterial activity of clove against E. coli was more effective at $50^{\circ}C$ than at $55^{\circ}C$.

• YAKHAK HOEJI
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• v.41 no.1
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• pp.126-131
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• 1997

To overcome the problems of the resistance to clavulanic acid, many researchers are developing novel inhibitors that are not sensitive to new mutant ${\beta}$-lactamases. In order to evaluate newly synthesized compound CH2150 (Sodium (3S.5R)-6(Z)-[1-{1-(2-{2-benzoxazoly}thioethyl)-l.2,3-txiazol-4-yl}methylene] penicillanate-1,1-dioxide) as a ${\beta}$-lactamase inhibitor, we examined inhibitory activity of CH2150 against ${\beta}$-lactamases of clinical isolates resistant to ampicillin/sulbactam(12 strains of Escherichia coli and 13 strains of Staphylococcus aureus), and compared with that of sulbactam. Nitrocefin was used as substrate for ${\beta}$-lactamases, and the increase of absorbance was measured spectrophotometerically at 482 nm. ${\beta}$-Lactarnase inhibition of CH2150 against ${\beta}$-lactamases was 73 ~ 96% in E. coli and 76 ~ 79% in S. aureus. Comparatively, that of sulbactam was 96 ~ 100% and 96 ~ 100%, respectively. The inhibitory activity of CH2150 was slightly lower than that of sulbactam. The MIC values of ampicillin combined with CH2150 (2:1) for the clinical isolates were 4~512 ${\mu}$g/ml for E. coli and 1.0 ~ 64 ${\mu}$g/ml for S. aureus, whereas 0.5~16 ${\mu}$g/ml for E. coli and 0.25~8 ${\mu}$g/ml for S. aureus when combined with sulbactam (2:1).

• Korean journal of food and cookery science
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• v.21 no.1 s.85
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• pp.84-93
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• 2005

The antibacterial activities of methanol extract and systematic solvent fractions( -hexane, chloroform, ethyl acetate, and butanol) from Korean common type figs at different ripening stages were tested by the broth dilution method against 8 representative food-poisoning bacteria- : L. monocytogenes, S. aureus, S. enteritidis, E. coli O157:H7, E. coli, Y. enterocolitica, V. parahaemolyticus, and S. typhimurium. The methanol extracts of unripened I and II showed stronger activity than that of the ripened figs especially against L. monocytogenes, S. enteritidis, E. coli O157:H7, Y. parahaemolyticus and S. typhimurium in 10 mg/mL. The systematic solvent fractions showed stronger antibacterial activities than the methanol extract, even al the lower concentrations. The hexane fraction of ripened figs showed higher growth inhibition than those of unripened I and II against L. monocytogenes, E. coli O157:H7, Y. enterocolitica and V. parahaemolyticus. The chloroform fraction showed strong antibacterial activity in all ripening stages against E. coli O157:H7 and V. parahaemolyticus. The butanol fraction showed better inhibition activity in unripened I and II than in the ripened figs. The hexane and chloroform fractions showed inhibition activity of more than $75\%$ against E. coli O157:H7, V. parahaemolyticus in 0.5 mg/mL. Each fraction showed a little different antibacterial activity according to the ripening stages of the fruits and the tested strains. Especially, figs in the unripened II stage revealed superior activity relatively and the hexane and chloroform fractions revealed the strongest activity, followed by the butanol fraction, while the ethylacetate and water fractions hardly showed any activity.

• Korean journal of food and cookery science
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• v.13 no.4
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• pp.440-447
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• 1997

The antibacterial effect of low concentrations of oregano (Origanum vulgare L.) in culture broth against Escherichia coli O157:H7 and Staphylococcus aureus 196E was tested at 35,5 and -20$^{\circ}C$. Tryptic soy broth (TSB) containing 0∼2% (w/v) of oregano was inoculated with 10$\^$6/∼10$\^$7/ CFU/$m\ell$ of E. coli or S. aureus and incubated at each temperature. The growth of E. coli was not inhibited at 0.1∼1.0% oregano and the growth occured at 2% oregano but only after a prolonged lag period. The death rate of E. coli after stationary phase was increased with increasing concentration of oregano in culture broth. The growth of S. aureus was inhibited with increasing concentration of oregano at 35$^{\circ}C$. Growth of S. aureus occured at the presence of 0.3∼0.5% oregano after a long lag period while the viability at 1.0∼2.0% was decreased during storage at 35$^{\circ}C$. During refrigerated storage at 5$^{\circ}C$, inhibition of E. coli or S. aureus was increased with the progress of time and increasing spice concentration. At the presence of 0.5∼2.0% oregano, E. coli and S. aureus were dead after 20 and 16 days of refrigerated storage, respectively. During frozen storage at -20$^{\circ}C$, the antibacterial activity of oregano against E. coli was increased with increasing storage time and spice concentration while the antibacterial activity against S. aureus was effective during the early period of storage, and no changes in the population of S. aureus occurred at different concentrations of oregano during frozen storage. Viable counts of E. coli were 1/3∼l/7 and S. aureus were 1/18∼l/46 of the control at 0.1% oregano in culture broth during frozen storage.

• Animal cells and systems
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• v.15 no.1
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• pp.29-36
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• 2011

Insect lysozymes are basic, cationic proteins synthesized in fat body and hemocytes in response to bacterial infections and depolymerize the bacterial cell wall. The c-type lysozyme of the insect Spodoptera litura (SLLyz) is a single polypeptide chain of 121 residues with four disulfide bridges and 17 rare codons and is approximately 15 kDa. The full-length SLLyz cDNA is 1039 bp long with a poly(A) tail, and contains an open reading frame of 426 bp long (including the termination codon), flanked by a 54 bp long 5' UTR and a 559 bp long 3' UTR. As a host for the production of high-level recombinant proteins, E. coli is used most commonly because of its low cost and short generation time. However, the soluble expression of heterologous proteins in E. coli is not trivial, especially for disulfide-bonded proteins. In order to prevent inclusion body formation, GST was selected as a fusion partner to enhance the solubility of recombinant protein, and fused to the amplified products encoding mature SLLyz. The expression vector pGEX-4T-1/rSLLyz was then transformed into E. coli BL21(DE3)pLysS for soluble expression of rSLLyz, and the soluble fusion protein was purified successfully. Inhibition zone assay demonstrated that rSLLyz showed antibacterial activity against B. megaterium. These results demonstrate that the GST fusion expression system in E. coli described in this study is efficient and inexpensive in producing a disulfide-bonded rSLLyz in soluble, active form, and suggest that the insect lysozyme is an interesting system for future structural and functional studies.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1238-1243
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• 2009

The susceptibilities of major enterohemorrhagic Escherichia coli (EHEC) strains to antimicrobial agents and the cytotoxicity of these agents were examined using a total of 38 strains of E. coli O26, O111, and O157, which are the major serogroups of EHEC. Among the 38 strains, 35, 36, and 36 were susceptible to amikacin, imipenem, and norfloxacin, respectively. These antimicrobial agents were further examined to determine their cytotoxicity on Vero cells as well as their effect on the release of Shiga toxins along with trimethoprim/sulfamethoxazole. Each of the E. coli O26, O111, and O157 strains containing both the stx1 and stx2 genes were grown in the absence or presence of these agents at 1/4 minimal inhibitory concentration for 6 h, 12 h, and 18 h. At the concentrations used in this study, none of the agents significantly altered cell count compared with the control group. The level of cytotoxicity in the imipenem group was lower at 12 hand 18 h than their respective controls. In contrast, the level of cytotoxicity in cultures treated with trimethoprim/sulfamethoxazole, norfloxacin, and amikacin was increased. The strains were also examined for the release of Shiga toxins 1 and 2 following treatment with the agents, which were measured by the reversed passive latex agglutination (RPLA) method. The RPLA assay showed a suppression of release of Shiga toxin 2 in the strain cultures containing imipenem. These results indicate that imipenem may be a safe and effective agent for inhibition of these bacteria, which has clinical implications for the treatment of EHEC infections.

• The Korean Journal of Food And Nutrition
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• v.18 no.1
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• pp.81-87
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• 2005

This study was performed to investigate the antimicrobial activities of the Plagiorhegama dubium extracts against food-borne pathogens. The methanol extract was partitioned into petroleum ether, chloroform, ethyl acetate, and methanol portions. The antimicrobial activity of the P. dubium extracts was determined using a paper disc method against food-borne pathogens and food spoilage bacteria. The methanol extracts of P. dubium showed the highest antimicrobial activity against Staphylococcus aureus and Escherichia coli. The synergistic effect has been found in combined extracts of P. dubium and Hedyotis diffsa as compared to each extracts alone. Finally, the growth inhibition curve was determined using methanol extracts of P. dubium against S. aureus and E. coli. The methanol extract of P. dubium showed strong antimicrobial activity against S. aureus at the concentration of 4,000 ppm. The 4,000 ppm of ethyl acetate portion from P. dubium retarded the growth of S. aureus more than 36 hours and E. coli up to 60 hours. The methanol extracts of P. dubium has been shown the antimicrobial activity against S. aureus and E. coli.

• Korean Journal of Microbiology
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• v.53 no.2
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• pp.118-122
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• 2017

Bacterial growth inhibition by lead, zinc and cadmium was measured by using modified Tris minimal medium. The toxicity against Escherichia coli strain was in the order of zinc> cadmium> lead, and the Escherichia coli strain overexpressing iron-containing superoxide dismutase 2 of Streptomyces coelicolor A3(2) was found to have resistance to heavy metals.

• Biomolecules & Therapeutics
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• v.6 no.4
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• pp.383-388
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• 1998

The present study was aimed to investigate the preventive effects of rebamipide on the multiple organ dysfunction in a rat model of circulatory shock induced by bacterial endotoxin (E. coli lipopolysaccharide; LPS) in comparison with that of methotrexate. Endotoxemia for 6 hours resulted in little change in the levels of hemoglobin and neutrophils. However, treatment with methotrexate decreased significantly the numbers of circulating neutrophils. Significant increases in serum alanine aminotransferase (ALT,958 $\pm$ 250 lU/L, p<0.001) and aspartate aminotransferase (AST, 1350 $\pm$ 295 lU/L, p<0.001) levels induced by endotoxemia were significantly decreased by rebamipide and methotrexate. The increased level of lactic acid dehydrogenase (LDH) by LPS (2850 $\pm$ 467 lU/L, p<0.05) was significantly inhibited by rebamipide, but not by methot.elate. The elevated serum creatinine (1.2$\pm$0.1, p.0.05) and urea levels (55.3$\pm$6.5 mg/dL, p.0.01) by LPS were also decreased by rebamipide, but not by methotrexate. In line with these results, the plasma concentration of tumor necrosis factor-$\alpha$ (TNF-7,167 $\pm$ 20 pg/mL) was significantly increased upon injection of endotoxin at 1 hour by 1570$\pm$100 pg/mL, and declined to 312$\pm$35 pg/mL at 6 hours. The TNF-$\alpha$ level at 6 hours was significantly decreased by rebamipide to 207$\pm$8 pg/mL (P<0.05). Taken together, it is summarized that rebamipide inhibits the development of multiple ogran dysfunction by inhibition of neutrophil activation in association with inhibition of TNF-$\alpha$ formation in a murine model of endotoxemia.