• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1015-1025
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• 2015

The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681. First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the L-leucine-specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A_6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A_6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A_6-D-Leu-domain with the A_6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives.

• The Journal of the Korean Society for Microbiology
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• v.6 no.1
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• pp.29-40
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• 1971

Various kinds of antibiotics are generally believed to have inhibitory effects on the antibody response. However, as polymyxin B which belongs to the cyclic polypeptide group of antibiotic was found to have some enhancing effects on the antibody response of rabbits to enterobacterial common antigen(CA) under specified conditions, experiments were carried out on this problem with the following results. 1. When mixture of polymyxin B and CA derived from Salmonella typhimurium(STM) was treated 30 minutes at $37^{\circ}C$ and injected three times into rabbits by intravenous route, the antibody response to CA was weaker than rabbits injected CA only. 2. Mixture of polymyxin B and CA showed a marked antibody production when injected into rabbits primed with small amounts of heat-extracted antigen of STM, while the injection of CA alone showed low titers of response. 3. Mixture of polymyxin B and heat-extracted CA-containing antigen of Escherichia coli 014 also showed a increased antibody production than CA alone in rabbits primed with antigen of STM. 4. The effect of polymyxin B appeared in different ways. This antibiotic did not enhance the CA antibody response in rabbits primed with small amounts of E. coli 0111 and 055, but enhance in rabbits primed with Shigella flexneri. 5. No enhancing effect on the antibody response was observed by polymyxin B in rabbits primed with CA. 6. No enhancing effect on the antibody response was also noted in rabbits primed with STM antigen in case polymyxin B and CA were administered simultaneously but in veins of different places. 7. Bacitracin did not enhance the CA antibody response in primed rabbits with STM antigen, but neomycin slightly enhance the response. 8. Lipopolysaccharide showed no priming effect on the CA antibody response, and no enhancement of the CA antibody response in rabbits printed with STM. 9. The priming effect of STM antigen against CA antibody response was very weak as compared with the effect of CA derived from STM antigen.

• Journal of Veterinary Clinics
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• v.27 no.1
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• pp.6-10
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• 2010

Bacterial pathogens were isolated from dogs with urinary tract infection (UTI) in local animal hospitals between August 2003 and December 2009. Bacteria were isolated from urine of 47 dogs. The isolated pathogens were Escherichia coli (n = 27), Streptococcus spp. (n = 7), Staphylococcus spp. (n = 5), Enterobacter spp. (n = 3), Proteus spp. (n = 2), other species were 3 strains, respectively. E. coli were susceptible to imimpenem, polymyxin B, amikacin, cephalosporins, aztreonam, amoxicillin clavulate, cephalosporins, tricarcillin, and amoxicillin clavulate, while were resistant bacitracin, erythromycin, lincomycin, oxacillin, penicillin, and novobiocin. Streptococcus spp. were susceptible to bacitracin, imimpenem, and trimethoprime-sulfa, while were highly resistant amikacin, cefotaxim, cefoxitin, cloxacillin, gentamicin, lincomycin, oxacillin, penicillin, streptomycin, and tobramycin. Staphylococcus spp. were susceptible to cefoxitin, doxycycline, enrofloxacin, imimpenem, and tobramycin, but were resistant aztreonam and tetracycline.

• Macromolecular Research
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• v.13 no.6
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• pp.549-552
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• 2005

• Journal of Life Science
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• v.14 no.4
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• pp.664-669
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• 2004

The contamination of Bacillus cereus was investigated in 240 RTE (ready-to-eat) food samples including 118 seafoods, 82 Korean packaged meals and 40 other RTE foods. Many B. cereus presumptive strains were isolated from the enrichment culture in Tryptic Soy Broth (TSB) added polymyxin, followed by selective culture in Mannitol Egg Yolk Polymyxin (MYP) agar and Gram staining. A total of 36 strains (16 in seafoods, 17 in Korean pack-aged meals and 3 in other RTE foods) were identified as B. cereus by the analysis of 61 biochemical tests of the API 50CHB/20E system test and supplementary tests of $\beta$-hemolysis, rhizoid growth, motility and oxidase activity. The 28 strains out of 36 B. cereus isolates produced diarrhoeal enter-otoxin in Brain Heart Infusion (BHI) broth. All isolates were resistant to ampicillin and penicillin antibiotics, and most of them were susceptible to gentamicin, vancomycin, bacitracin, chloram-phenicol, kanamycin and streptomycin. The growth of B. cereus was affected by environmental temperature and incubation time. Culture with temperature under 1$0^{\circ}C$ effectively restricted the growth of B. cereus.

• Toxicological Research
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• v.13 no.1_2
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• pp.111-116
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• 1997

In the present study, we investigated the neutralization activity of various antimicrobials against lipopolysaccharide (LPS) as well as interaction between two antimicrobials (enrofioxacin and coilstin) using checkerboard method. The neutralization activity of antimicrobials used in the test was assayed by means of LAL chromogenic test after reaction of LPS with colistin, enorfioxacin, ampicillin, polymyxin B, oxytetracycline, streptomycin, and erythromycin. As the results, the neutralization activity of coltstin and polymixin B had a more stronger than that of tested other antimicrobials. In bacterial culture broth, the best neutralization activity of the antibiotics was also shown to coltstin and polymixin B. Meanwhile, It was shown to have synergism between enorfloxacin and coltstin on the basts of FIC (fractional inhibition concentration). The FIC of enorfioxacin-colistin combinations was 0.50-1.03 to Staphylococcus aureus R-209, 1.03-1.06 to Salmonella typhimurium, 0.75-1.25 to Bordetella Bronchtseptica and 1.02-1.25 to E. coli K88ab. On the basts of the above results, the present study may be of clinical usefulness in the choice of an antibiotic therapy for severe sepsts in animals.

• International Journal of Oral Biology
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• v.30 no.1
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• pp.31-37
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• 2005

Treponema maltophilum, a Group IV oral spirochete, is associated with periodontitis and endodontic infections. In this study we analyzed the functional role of the major surface protein of this organism (MspA) in human gingival fibroblasts (HGFs). The full-length gene encoding MspA was cloned and expressed in Escherichia coli by using the expression vector pQE-30. The recombinant protein (rMspA) was purified by affinity chromatography with nickel-nitrilotriacetic acid agarose and possible contamination of E. coli endotoxin in rMspA was removed by using polymyxin B-agarose. rMspA significantly induced the expression of pro inflammatory cytokines like IL-6 and IL-8 and intercellular adhesion molecule (ICAM)-1 in HGFs, when analyzed by reverse transcription-PCR, flow cytometry, and enzyme-linked immunosorbent assay. Our results indicate that MspA of T. maltophilum may play an important role in amplifying the local immune response by upregulating the expression of proinflammatory cytokines and ICAM-1.

• Korean Journal of Veterinary Research
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• v.61 no.2
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• pp.16.1-16.6
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• 2021

Pseudomonas aeruginosa is one of the most common pathogenic species associated with canine otitis externa (OE). Their resilience is achieved by forming a biofilm, which allows these bacteria to evade even the harshest of treatments. This study evaluated the in vitro synergistic efficacy of N-acetylcysteine (NAC) with different antimicrobial agents against P. aeruginosa isolated from dogs with OE to develop an effective treatment against P. aeruginosa. The antimicrobial activity was evaluated by the minimum inhibitory concentration test using the microdilution method. The efficacy of antibiofilm formation was evaluated using a crystal violet stain method. The treatment solutions included NAC alone, and in synergy with enrofloxacin, polymyxin B, and gentamicin. NAC alone exhibited antimicrobial and antibiofilm abilities. On the other hand, the combination of NAC and the antibiotics did not show any significant synergistic effects against P. aeruginosa.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.39-46
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• 1983

We collected three hundred thirty-five strains of Salmonella typhi isolated from human sources during the period January to December 1982 Korea. Most of them were from general hospitals and city health center, the remaining one hundred sixtyone strains were obtained from other 12 provincial health centers among 335 testing strains. We used ninety-nine Vi-phages as distributed from International Center for Enteric phage Typing(ICEPT) in London. We found nine phage types among 335 strains of Salmonella typhi in this study. Additionally, I+IV, degraded Vi-positive and Vi negative strains were presented. This study documented the occurrence of the new $B_2$ type in Korea for the first time. The present basic phage type formula in Korea appeared to be A, $B_2,\;D_1,\;D_2,\;D_4,\;D_6,\;D_8,\;D_{12},\;E_1,\;E_9,\;D_1,\;M_1$, 40 and plus I+IV and degraded vi strains(Table 2). The current phage types of Salmonella typhi isolated in Korea 1982 were A, $B_2,\;D_1,\;D_2,\;D_6,\;D_8,\;E_1,\;M_1$ and 46 $M_1$ type was widely distributed all over the country, and E type was next predominant. Antibiotic susceptibility test were performed by means of Kirby-Bauer disc diffution method using 12 kind of antibiotics such as Ampicillin, Carbenicillin, Cephalosporin, Chloramphenicol, Colistin, Gentamicin, Kanamycin, Nalidix acid, Neomycin, Polymyxin-B, Streptomycin and Tetracycline. The sensitivity pattern to antibiotics of Salmonelia typhi cultures were summarized.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.5
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• pp.798-802
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• 2009

This study was performed to assess the presence of contaminated microorganisms of Escherichia coli, Clostridium perfringens, and Bacillus cereus in the 112 commercial Saengshik products. E. coli was not detected in all the samples, but C. perfringens was detected in 11 products (9.8%). The number of the bacteria was less than 100 CFU/g, which was satisfactory to KFDA microbiological requirement. B. cereus was detected less than $10^2{\sim}10^3$ CFU/g in 7 products and $10^3{\sim}10^4$CFU/g in 13 products out of 25 products. Those detected bacteria from tryptose sulphite cycloserine agar and mannitol egg yolk polymyxin agar showed the typical characteristics of Gram positive and contained lecithinase, which can decompose egg-yolks layers in the biochemical test. Therefore, much more attention must be applied to satisfy the B. cereus requirement for Saengshik products sold in the market.