• Journal of Yeungnam Medical Science
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• v.30 no.2
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• pp.83-89
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• 2013

Background: The prevalence of extended-spectrum ${\beta}$-lactamase (ESBL)-producing Escherichia coli is increasing rapidly worldwide. Treatment options for ESBL-producing E. coli are limited, and infections caused by this organism are associated with improper antibiotic use, a long hospital stay, and increased mortality. Thus, the assessment and early recognition of the risk factors of nosocomial infections due to ESBL-producing E. coli are important for the infection control and proper treatment. Methods: A case-control study was performed that included nosocomial episodes of ESBL-producing E. coli bacteremia at a tertiary care hospital from January 2004 to December 2007. For each case patient, three controls were randomly selected and data on predisposing factors were collected. Results: Fifty-five cases of nosocomial ESBL-producing E. coli bacteremia were studied. Carbapenem usage (OR: 11.3, 95% CI: 1.1-115.9, p=0.041), quinolone usage (OR: 4.5, 95% CI: 1.1-18.8, p=0.042), biliary obstructive disease (OR: 11.8, 95% CI: 3.0-46.7, p<0.001) and the APACHE II score (OR: 1.3, 95% CI: 1.2- 1.5, p<0.001) were analyzed as independent risk factors of nosocomial ESBL-producing E. coli bacteremia. Conclusion: Our results showed that physicians caring for patients with risk factors of nosocomial bacteremia should consider ESBL-producing E. coli as the causative organisms of the disease.

• Biomedical Science Letters
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• v.12 no.3
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• pp.267-274
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• 2006

Klebsiella pneumoniae is the leading cause of nasocomial infection and the most commonly isolated from clinical specimens. $Extended-spectrum-{\beta}-lactamase$ (ESBL) producing K. pneumoniae infection was associated with a significantly longer duration of hospital stay and greater hospital charges. The purpose of this study is to investigate the antibiotic resistant patterns and the DNA fingerprint types of extended-spectrum ${\beta}-lactamase$ (ESBL) producing K. pneumoniae. 223K. pneumoniae strains were collected from three general hospitals with more than 500 beds in Busan, Korea from September 2004 to October 2005. The minimum inhibitory concentration (MIC) of antibiotics was measured using the Gram negative susceptibility (GNS) cards of VITEK (Vitek system, Hazelwood Inc., MO). Random amplified polymorphic DNA method was used to detect DNA fingerprint of the organisms. Of the 226 K. pneumoniae isolates 65 ESBL-producing K. pneumoniae strains were detected by the Vitek system and confirmed by the double-disk synergy test. All the 65K. pneumoniae strains were resistant cefazolin, cefepime, ceftriaxone and aztreonam, and 83.0% of the organisms were resistant to ampicillin/sulbactam, 66.1% to tobramycin, 67.6% to piperacillin/tazobactam, 61.5% to ciprofloxacin, and 47.6% to trimethoprim/sulfamethoxazole and 43.0% to gentamicin. The RAPD patterns were distincted as 10 types by three random 10-mer primers (208, 272, 277). Among ten type patterns, three types (Ic, IIb, IIIe) were remarkably represented at patient of internal department, nerve surgery department, general surgery department, and neonatal room. These results indicate that RAPD can be useful for DNA of strains typing in the epidemiological investigations. Therefore more investigation are needed in order to prevent the ESBL type-producing K. pneumoniae from spreading resistance to oxyimino cephalosphorin antibiotics.

• 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 Microbiology
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• v.45 no.2
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• pp.163-169
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• 2009

This study was performed to investigate the type of extended-spectrum $\beta$-lactamases (ESBL) produced by bacteria isolated from the sewage of wastewater treatment plant at Minragdong, Suyong-gu in Busan. The facility is located at sushi restaurants and guides its drain water to the wastewater treatment plant at Yonghodong, Nam-gu in Busan. Samples were collected on January, 2009. A total of 19 strains were selected as potential ESBL positive strains through a double disk synergy test. On the basis of the results from biochemical tests including indole, methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylase-dihydrolase and sugar-fermentation tests, the 19 strains were identified with 16 strains of Escherichia coli and 3 strains of Klebsiella pneumoniae. Out of 19 strains, 4 transconjugants against Escherichia coli J53, which is sodium azide resistant recipient strain, were obtained. The plasmids isolated from transconjugants were used for PCR analysis. The type of each extended-spectrum $\beta$-lactamase (ESBL) produced by the strains was determined on the basis of isoelectric focusing analysis and DNA sequencing. The results indicated that the types of ESBL from Klebsiella pneumoniae were SHV-12 (3 strains), and Escherichia coli was SHV-12/TEM-1 (1 strain), respectively.

• The Korean Journal of Mycology
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• v.8 no.1
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• pp.63-68
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• 1980

${\beta}-Lactamase$ was isolated from the culture filtrate of the penicillin producing strain, Penicillium chrysogenum Q176. When the pH of the medium was adjusted to 5.0 at the start of culture, a rapid increase in pH accompanied by the synthesis of penicillin was observed in the first $2{\sim}4$ days. When the pH of medium was brought to 6.0 or 7.0 the opposite was observed: high yield of the enzyme and little of the antibiotics in the medium. The optimum enzyme activi­ty was at a temperature of $40^{\circ}C$ and around pH 7.0. A partially purified enzyme was assayed on several different substrates including penicillins V and G, 6-aminopenicillanic acid, cephalospo­rin C. The V max values calculated were 24.5, 20.4, 7.6, and 6.1 mmoles/hour, and the $K_m$, values were 16.4, 12.6, 7.5, and 6.9 mM in the order given.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1884-1889
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• 2008

The ${\beta}$-lactamase inhibitory protein, BLIP-II, found in the culture supernatant of Streptomyces exfoliatus SMF19, shows no discernible sequence identity with other ${\beta}$-lactamase inhibitory proteins identified in Streptomyces spp. A null mutant of the gene encoding BLIP-II (bliB::$hyg^r$) showed a bald appearance on solid media. Although BLIP-II was initially isolated from the supernatant of submerged cultures, sites of BLIP-II accumulation were seen in the cell envelope. Mutation of bliB was also associated with changes in the formation of septa and condensation of the chromosomal DNA associated with sporulation. The bliB mutant exhibited infrequent septa, showing dispersed chromosomal DNA throughout the mycelium, whereas the condensed chromosomes of the wild-type were separated by regularly spaced septa giving the appearance of a string of beads. Therefore, on the basis of these results, it is suggested that BLIP-II is a regulator of morphological differentiation in S. exfoliatus SMF19.

• Molecules and Cells
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• v.27 no.2
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• pp.225-235
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• 2009

Antibody phage display provides a powerful and efficient tool for the discovery and development of monoclonal antibodies for therapeutic and other applications. Antibody clones from synthetic libraries with optimized design features have several distinct advantages that include high stability, high levels of expression, and ease of downstream optimization and engineering. In this study, a fully synthetic human scFv library with six diversified CDRs was constructed by polymerase chain reaction assembly of overlapping oligonucleotides. In order to maximize the functional diversity of the library, a ${\beta}$-lactamase selection strategy was employed in which the assembled scFv gene repertoire was fused to the 5'-end of the ${\beta}$-lactamase gene, and in-frame scFv clones were enriched by carbenicillin selection. A final library with an estimated total diversity of $7.6{\times}10^9$, greater than 70% functional diversity, and diversification of all six CDRs was obtained after insertion of fully randomized CDR-H3 sequences into this proofread repertoire. The performance of the library was validated using a number of target antigens, against which multiple unique scFv sequences with dissociation constants in the nanomolar range were isolated.

• Applied Biological Chemistry
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• v.40 no.1
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• pp.23-29
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• 1997

To develop novel cephem antibiotics, We have synthesized a new compound, named YH-487, by attaching the thiol and aminothiazole residue to $C_3$ and $C_7$ position of 7-ACA, respectively. Several characteristics such as structure, antibiotic spectrum, action mechanism, stability against ${\beta}-lactamase$ and synergistic effect were investigated. Anti-bactericidal activity of YH-487 against gram-positive and gram-negative bacteria were superior to that of the other cephem antibiotics. We have examined the action mechanisms of YH-487 using penicillin binding protein (PBP) assay, and found that the bactericidal activity was obtained by inhibiting PBP-1A, PBP-1B and PBP-3. YH-487 showed synergistic effect with gentamicin, tobramycin, and amikacin against Pseudomonas aeruginosa. In addition, YH-487 was effective against Enterobacter cloacae in combination with amikacin. Based on the above observations, YH-487 was classified as a novel third-generation ${\beta}-lactam$ antibiotics.

• Microbiology and Biotechnology Letters
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• v.51 no.3
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• pp.309-313
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• 2023

Here we report essential features of the draft genome of an AmpC-β-lactamase-producing bacterial isolate obtained from farm tomatoes in South Africa. The isolate designated strain Tom1 featured a genome of 4950426 bp with a G+C% of 59.83. It was identified as Serratia marcescens by ribosomal multilocus sequence typing (rMLST), digital DNA-DNA hybridization (dDDH), average nucleotide identity (ANI), and phylogenetic analysis using reference genomes. Its genome encoded an AmpC-β-lactamase (bla_SST-1), an efflux pump providing tetracycline resistance (tet(41)), and an aminoglycoside acetyltransferase (aac(6')-Ic). Additionally, genes encoding proteins involved in prodigiosin biosynthesis and associated with adherence, biofilm formation, virulence, and pathogenicity were detected.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.20-27
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• 1994

An inducible ${\beta}$-lactamase gene (bla) was identified and isolated from the chromosomal DNA of multiple drug resistant strains of Staphylococcus aureus. Determined base sequence of bla and of its flanking region was compared with those of bla genes identified on the staphylococcal plasmids pPC1, pI258, pI1071, and pUB101. Base sequence of 843 base-long structural gene of our bla was same as that of pPCl-, pI258-, and pS1-bla. However, HindIII recognition site Which is found in most of the bla genes at 140 base upstream from the structural gene was moved to the site of 370 base upstream from the structural gene. And one of the two direct repeat sequence found in downstream flanking region of pI1071-bla was deleted in our bla. Amino acid sequence homology analysis of the ORF located around HindIII recognition site reveals that this 80 amino acids-long polypeptide is C-terminus of transposase of Tn4001.