• KSBB Journal
• /
• v.6 no.1
• /
• pp.111-114
• /
• 1991

The effects of the precursor processing inhibitor, carbonylcyanide-chlorophenyl hydrazone(CCCP), are investigated on the production of soluble ${\beta}-lactamase$ the formation of the inclusion body when ${\beta}-lactamase$ is overproduced by induction with isopropyl thiogalactoside(IPTG). When cells are treated by CCCP, more soluble ${\beta}-lactamase$ is produced. In this case, no difference in the amount of inclusion body is observed.

• Microbiology and Biotechnology Letters
• /
• v.16 no.5
• /
• pp.398-401
• /
• 1988

Streptomyces sp. producing $\beta$-lactamase inhibitor were isolated from soil. The culture conditions for the production of the $\beta$-lactamase inhibitor were evaluated and isolation produce of the $\beta$-lactamase inhibitor from the culture broth was also established. Some characters of the partially purified $\beta$-lactamase were determined.

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

• Journal of Microbiology and Biotechnology
• /
• v.1 no.2
• /
• pp.85-89
• /
• 1991

The aim of this study is to elucidate characteristics of ${\beta}-lactamase$ inhibitor produced by Streptomyces sp. SMF-19 isolated from soil was found to produce proteinaceous extracellular ${\beta}-lactamase$ inhibitor. The ${\beta}-lactamase$ inhibitor was purified through ammonium sulfate fractionation, gel filtration, anion exchange chromatography and fast performace liquid chromatography. The molecular weight of the ${\beta}-lactamase$ inhibitor was estimated to be about 48,000 by SDS-PAGE. The mode of inhibition against penicillin G as a substrate was uncompetitive. The ${\beta}-lactamase$ inhibitor was stable in wide pH range but unstable at high temperature above $50^{\circ}C$.

• Microbiology and Biotechnology Letters
• /
• v.19 no.5
• /
• pp.439-443
• /
• 1991

A strain of Streptomyces sp. isolated from soil was found to produce extra-cellular $\beta$-lactamase associated partially to the cell growth. The $\beta$-lactamase was purified from the culture supernatant through anmonium sulfate fractionation, ion-exchange chromatographies and gel filtration. The final purification fold and recovery yield were 57 and 6.2%, respectively. Molecular weight of the $\beta$-lactamase was estimated to be about 67, 000 by SDS-polyacrylamide gel electrophoresis. The optimal reaction condition was at pH 7-8 and at 35-$45^{\circ}C$. The $K_m$ and $V_{max}$ values of the enzyme for penicillin G were estimated to he 3 mM and $3\times 10^3$ $\mu\textrm{M}$/min/mg protein, respectively. The purified $\beta$-lactamase was classified to the class A enzyme hydrolyzing only penicillin.

• YAKHAK HOEJI
• /
• v.51 no.6
• /
• pp.447-454
• /
• 2007

The resistance of bacteria against ${\beta}$-lactam antibiotics is mainly caused by the production of ${\beta}$-lactamase enzymes. ${\beta}$-Lactamase inhibitors are used in combination with known antibiotics to overcome the growing problem of bacterial resistance. We prepared 6-(thienylmethylene)penam sulfones for the development of potent ${\beta}$-lactamase inhibitors and evaluated their ${\beta}$-lactamase inhibitory activities.

• Biomolecules & Therapeutics
• /
• v.14 no.4
• /
• pp.220-225
• /
• 2006

Derivatives of penicillanic acid sulfones are known to be irreversible inhibitors of $\beta$-lactamase. Eight 6-tricyclic methylene penicillanic acid sulfones were prepared, and their $\beta$-lactamase inhibitory activities were evaluated against $\beta$-lactamase types I, II, III and IV. Among the tricycles attached to 6-exomethylenepenam sulfones, thiazolobenzimidazole(12a-12b), fluorene(12c), and carbazole(12e), showed inhibitory activity on type I, II and III $\beta$-lactamase. But phenanthrene(12d), and anthracene(12f-12h) derivatives showed little $\beta$-lactamase inhibitory activity. The synergic effects of the selected compound(l2b) in 1:4 combination with piperacillin showed some protection to piperacillin for the resistant strains of E. coli DC2 and P. aeruginosa 1771.

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

• Journal of Microbiology and Biotechnology
• /
• v.22 no.4
• /
• pp.555-562
• /
• 2012

PCR was performed to analyze the ${\beta}$-lactamase genes carried by ampicillin-resistant Vibrio spp. strains isolated from marine environments in Korea between 2006 and 2009. All 36 strains tested showed negative results in PCR with the primers designed from the nucleotide sequences of various known ${\beta}$-lactamase genes. This prompted us to screen new ${\beta}$-lactamase genes. A novel ${\beta}$-lactamase gene was cloned from Vibrio alginolyticus KV3 isolated from the aquaculture water of Geoje Island of Korea. The determined nucleotide sequence (VAK-3 ${\beta}$-lactamase) revealed an open reading frame (ORF) of 852 bp, encoding a protein of 283 amino acids (aa), which displayed low homology to any other ${\beta}$-lactamase genes reported in public databases. The deduced 283 aa sequence of VAK-3, consisting of a 19 aa signal peptide and a 264 aa mature protein, contained highly conserved peptide segments specific to class A ${\beta}$-lactamases including the specific amino acid residues STFK (62-65), SDN (122-124), E (158), and RTG (226-228). Results from PCR performed with primers specific to the VAK-3 ${\beta}$-lactamase gene identified 3 of the 36 isolated strains as V. alginolyticus, Vibrio cholerae, and Photobacterium damselae subsp. damselae, indicating the utilization of various ${\beta}$-lactamase genes including unidentified ones in ampicillin-resistant Vibrio spp. strains from the marine environment. In a mating experiment, none of the isolates transfered the VAK-3 ${\beta}$-lactamase gene to the Escherichia coli recipient. This lack of mobility, and the presence of a chromosomal acyl-CoA flanking sequence upstream of the VAK-3 ${\beta}$-lactamase gene, led to the assumption that the location of this new ${\beta}$-lactamase gene was in the chromosome, rather than the mobile plasmid. Antibiotic susceptibility of VAK-3 ${\beta}$-lactamase was indicated by elevated levels of resistance to penicillins, but not to cephalosporins in the wild type and E. coli harboring recombinant plasmid pKV-3, compared with those of the host strain alone. Phylogenetic analysis showed that VAK-3 ${\beta}$-lactamase is a new and separate member of class A ${\beta}$-lactamases.

• Korean Journal of Veterinary Research
• /
• v.43 no.1
• /
• pp.113-120
• /
• 2003

The antimicrobial effect of ${\beta}$-lactam antibiotics, which had ${\beta}$-lactamase inhibitor activity, on Staphylococcus aureus isolated from mastitis was investigated in this study. Out of 166 isolates, 99 isolates (59.6%) produced ${\beta}$-lactamase, and 98 isolates of 99 were ${\beta}$-lactamase positive in above $12.5{\mu}g/m{\ell}$ MIC of penicillin. In the providence distribution, ${\beta}$-lactamase production rate of 4 providence, Gangwon, Gyeonggi, Chungcheong, and Jeolla was 100%, 65.7%, 58.8%, and 50.0%, respectively. Antibiotic activities of ${\beta}$-lactam antibiotics against lactamase positive isolates also were investigated. Antimicrobial effects of ampicillin/sulbactam or amoxicillin/clavulanic acid treated group were better than ampicillin or amoxicillin treated group. In antimicrobial effects on intracellular S aureus, there was no difference 1 hour and 4 hour treatment in control, ampicillin, and amoxicillin group, but in 18 hours treatment, ampicillin/sulbactam or amoxicillin/clavulanic acid had a better effect than ampicillin or amoxicillin (p<0.05).