• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.6
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• pp.597-604
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• 2011

Using 108 strains of Vibrio parahaemolyticus isolated from seawater, we investigated ampicillin-resistance profiles and the genetic characterization of ${\beta}$-lactamase (VPA0477). All of the strains studied, except one strain, were resistant to ampicillin. However, the strain that was susceptible to ampicillin had the same ${\beta}$-lactamase gene as the ampicillin-resistant strains. We compared ${\beta}$-lactamase promoter region sequences among five strains, including both ampicillin-resistant and -susceptible strains. In the susceptible strain, a nucleotide at position -19 in the methionine initiation codon for ${\beta}$-lactamase was not present in the ampicillin-resistant strains. The genes in the region containing the gene VPA0477 were present in all of the tested strains, and LA-PCR analysis showed that the distance between VPA0474 and VPA0479 in all of the V. parahaemolyticus samples was precisely 5.7 kb. In V. parahaemolyticus ${\beta}$-lactamase, four important structural features that are conserved in Class A ${\beta}$-lactamases were present in the deduced amino acid sequences. Taken together, our study demonstrates that V. parahaemolyticus ${\beta}$-lactamase is included in the Class A ${\beta}$-lactamase group, and some nucleotides within the promoter region are of particular importance for ${\beta}$-lactamase activity.

• YAKHAK HOEJI
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• v.41 no.4
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• pp.462-472
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• 1997

In this approach, the antimicrobial activities of the compounds were compared with the ${\beta}$-lactam antibiotics against ${\beta}$-lactamase producing strains in vitro. Heteroc yclyl exomethylenepenam derivatives were several numbers of 6-exomethylenepenam sodiums (CH1240, CH1245, CH1250, CH2140, CH2145, CH2150). The inhibitory concentraion assay of six compounds were compared with clavulanic acid, sulbactam, tazobactam. Clavulanic acid, sulbactam and tazobactam are used as inhibitors of a variety of plasmid-mediated beta-lactamases. In vitro ${\beta}$-lactamase inhibitory assay, CH1240 and CH2140 were more active than clavulanic acid, sulbactam and tazobactam against ${\beta}$-lactamases overally. And in vitro comparative antimicrobial susceptibility test of six inhibitors were performed with mixed forms of ampicillin, cefotaxime, amoxicillin, ticarcillin, piperacillin, cefoperazone against ${\beta}$-lactamase producing 31 species strains. Consequently CH2140 and CH1240 among the six compounds enhanced the activity of the ${\beta}$-lactams for 31 ${\beta}$-lactamase producing strains.

• YAKHAK HOEJI
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• v.41 no.5
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• pp.658-665
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• 1997

Identification of a soil microorganism strain X-8, producer of ${\beta}$-lactamase inhibitor, based on its morphological, physiological, biochemical and chemotaxonomical characteristics was performed. The strain X-8 was identified as Pseudomonas sp. The beta-lactamase inhibitor produced by this strain was highly achieved in fermentation medium contained glucose 0.5%, urea 0.25%, $K_2HPO_4{\cdot}3H_2O\;0.5%,\;MgSO_4{\cdot}7H_2O\;0.5%,\;FeSO_4{\cdot}7H_2O\;0.01%,\;CuSO_4,\;ZnSO_4,\;MnSO_4\;0.02%$. The beta-lactamase inhibitor was not extracted by organic solvent such as n-butanol and ethyl acetate but remained in aqueous layer. The n-butanol extract showed antimicrobial activity against M. smegmatis. The ${\beta}$-lactamase inhibitor was stable at pH 7.0~8.0 and 4$^{\circ}C$ for 24h. The ${\beta}$-lactamase inhibitor was bound on ion exchanger Diaion WA-30 and HP-20 and eluted with 2N-$NH_4OH$ and acetone, respectively.

• Bulletin of the Korean Chemical Society
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• v.14 no.5
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• pp.631-635
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• 1993

The polarizability and the transition state energy of a cephalosporin are assumed to be theoretical indices of the permeability through the outer membrane and of reactivity of ${\beta}$ -lactam ring with penicillin binding proteins, respectively, in Gram-negative bacteria. They are computed by AM1 method and used as variables of quantitative structure-activity relationship study. The results justify quadratic dependence of the activity on the variables. The intersection of difference volumes between $\beta-lactamase$ stable cephalosporins and unstable ones manifests that the steric hindrance of 7-side chain is responsible for the ${\beta}$ -lactamase stability.

• Journal of Life Science
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• v.18 no.11
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• pp.1592-1599
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• 2008

The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

• YAKHAK HOEJI
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• v.49 no.1
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• pp.51-55
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• 2005

In vitro ${\beta}$-lactamase inhibitory activity of 6-triazole exomethylenepenam compounds (1, 2, 3, 4, 5 and 6) was compared with clavulanic acid, sulbactam and tazobactam. The inhibitory activity of 3, 4 and 5 was stronger than those of sulbactam, clavulanic acid and tazobactam against Type IV enzymes. And, inhibitory activity of 3 and 4 was stronger than those of sulbactam, clavulanic acid and tazobactam against Type III enzymes. The in vitro antimicrobial activity of 3, 4 and 5 combined with ampicillin was better than those with sulbactam and with cefoperazone was compared with the sulbactam against ${\beta}$-lactamase producing 27 strains. The synergistic activity of (Z)-form compounds (3 and 5) was better than that of (E)-form compound (4) and sulfone compound (5) was better than sulfide compound (3).

• YAKHAK HOEJI
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• v.34 no.2
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• pp.106-111
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• 1990

The isolation and identification of an antibacterial component, from the leaves of Liriodendron tulipifera. K. Kotch against a cariogenic bacterium Streptococcus mutans OMZ 176, were carried out for developing of anticariogenic agents. The bioactive component was elucidated as ${\beta}-liriodenolide$, which was isolated newly from the leaves of L. tulipifera. The minimal inhibitory concentration (MIC) of ${\beta}-liriodenolide$ was $100\;{\mu}g/ml$ and the antibacterial activity was stronger than that of berberine. ${\beta}-Liriodenolide$ inhibited ${\beta}-lactamase$ activity, 50, 100 and $200\;{\mu}M$ ${\beta}-liriodenolide$ did ${\beta}-lactamase$ activity as 0.7, 3.5 and 19.7%, respectively. The toxicity of ${\beta}-liriodenolide$ was not found with the method of photohemolysis.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.590-596
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• 2001

A new extended-spectrum ${\beta}-lactamase$ with an isoelectric point (pl) of 6.2 was detected in Klebsiella pneumoniae Fl 61 that was isolated from a patient with infection. This strain was highly resistant to the third or fourth generation cephalosporins such as cceftazidime ceftriaxone, cefoperzaone, and cefpirome. Analysis of this strain by the double disk diffusion test showed synergies between amoxicillin-clavulanate (AMX-CA) and cefotaxime, and AMX-CA and aztreonam, which suggested that this strain produced a extended-spectrum ${\beta}-lactamase$ (ESBL). Cenetic analysis revealed that the resistance was due to the presence of a 9.4-kb plasmic, designated as pkpl 61, encoding for new ${\beta}-lactamase$ gene (bla). Sequence analysis showed that a new bla gene of pkpl 61 differed from $bla_{TEM-1}$ by three mutations leading to the following amino acid substitutions: $Val_{84}{\rightarrow}lie,{\;}Ala_{184}{\rightarrow}Val,{\;}and{\;}Gly_{238}{\rightarrow}Ser$. These mutations have not been reported previously in the TIM type ${\beta}-lactamases$ produced by clinical strains. The novel ${\beta}-lactamase$ was overexpressed in E. coli and purified by ion exchange chromatography on Q-Sepharose and CM-Sepharose, and then further purified by gel filtration on Sehadex G-200. The catalytic activity of th8 purified ${\beta}-lactamase$ was confirmed by the nitrocefin disk.

• Korean Journal of Microbiology
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• v.30 no.3
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• pp.149-153
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• 1992

Cloning of the gene encoding extracellular .betha.-lactamase from Streptomyces sp. SMF13 in a plasmid pIJ702 and expression of the gene in Streptomyces invidans were carried out. Optimal conditions for the formation of protoplasts of S.lividans and the regeneration of the protoplasts were evaluated. Streptomyces sp. SMF-13 was selected as a donor strain of .betha.-lactamase gene and totla DNA of the strain was partially digested with Sau3A I. DNA fragments ranged from 4kb to 10 kb were ligated to pIJ702 AT Bgl II site and then the ligated DNAs were transformed to the protoplasts of S, livivans. The transformation efficiency was $2 *10^{3}$ .$\mu$g DNA for the ligated DNA mixture. One colony among a thousand colonies regenerated showed extracellular .betha.-lactamase and the size of the inserted DNA fragment was estimated to be 3.94 kb. The .betha.-lactamase activity in the culture broth of the recombinant strain was maximum at 3 days culture to be 1.0 unit/ml.

• YAKHAK HOEJI
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• v.28 no.2
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• pp.89-95
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• 1984

Microorganisms having beta-latamase inhibitory activity were selected from soil samples collected from 63 spots throughout the country. Screening procedures consist of two steps. Those are growth inhibition test of penicillinase-producing Staphylococcus aureus by double-layered agar plate containing penicillin G as a substrate, and that of penicillin sensitive Staphylococcus aureus ATCC 6538 in the similiar condition including penicillinase. Finally, a strain was selected from a soil sample of Pa-ju, Kyeong-gi Do. This strain was classified as a Streptomyces sp. by ISP(International Streptomycete Project) and Bergey's manual.