• Title/Summary/Keyword: Lactamase

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Prevalence of Strains Resistant to the Third Generation Cephalosporins among Clinical Isolates and Identification of TEM Type $\beta$-lactamase from Resistant Strains by PCR Method (3 세대 세파계 항생제에 내성인 임상균주의 분포와 PCR 법을 이용한 TEM type $\beta$-lactamase 생산균주의 동정)

  • 김무용;오정인;송혜경;백경숙;곽진환
    • YAKHAK HOEJI
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    • v.39 no.3
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    • pp.276-282
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    • 1995
  • Compared to the first and second-generation cephalosporins, the third-generation cephalosporins are remarkably stable against hydrolysis by the $\beta$-lactamases produced by aerobic gram-negative bacilli, such as Enterobacteriaceae. Among these bacteria, the most prevalent plasmid-encoded $\beta$-lactamase is TEM-1 $\beta$-lactamase belonging to class A or group 2b. This enzyme is produced constitutively and is principally active against peniciflins and old cephalosporins rather than third-generafion cephalosporins, carbapenems and mmobactams. However, new TEM type $\beta$-lactamases including TEM-9 and TEM-12 evolved through point mutations in a gene encoding $\beta$-lactamase have been discovered from patients during chemotherapy. These $\beta$-lactamases are known to be capable of hydrolyzing most of the third-generatim cephalosporins. To study the prevalence of $\beta$-lactamases from clinical isolates collected in Korea. the minimal inhibitory concentratims(MICs) of several third-generation cephalosporins against 628 clinical isolates were determined by agar dilution methods, and $\beta$-lactamas-producing bacteria were isolated by use of cefinase disc. By polymerase chain reaction (PCR) method, clinical isolates harboring a gene for TEM type $\beta$-lactamase were identified among the $\beta$-lactamase producing strains. Twentiy three percent of the clinical isolates was resistant to the thirdgeneration cephalosporins, and more than 90% of resistant cells produced various $\beta$-lactamases. TFM type $\beta$-lactamases were dominant in gram-negative bacilli, such as Escherichia coli, Klebsiella pneumoniae, Enterobacter species. These results suggest the necessity of the development of new cephalosporins which are stable against $\beta$-lactamases like TEM.

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A Novel Plasmid-Mediated ${\beta}-lactamase$ that Hydrolyzes Broad-Spectrum Cephalosporins in a Clinical Isolate of Klebsiella pneumoniae

  • Kwak, Jin-Hwan;Kim, Mu-Yong;Chol, Eung-Chil
    • 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.

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Secretion of escherichia coli $\beta$-lactamase from bacillus subtilis with the aid of usufully constructed secretion vector

  • Park, Geon-Tae;Rho, Hyun-Mo
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.60-64
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    • 1992
  • The secretion vector with promoter and signal sequence region of neutral protease gene (npr) from Bacillus amyloliquefaciens was constructed by the technique of polymerase chain reaction (PCR). A unique restriction iste was introduced into the 3' of the signal coding region by the synthesis of PCR primer. To demonstrate the function of cloned promoter and signal sequence, we used the E. coli .betha.-lactamase structural gene as a foreign gene. The signal sequence of .betha.-lactamase gene was deleted by Bal31 exonuclease and only mature region was introduced into the secretion vector. Bacillus subtilis cells transformed by the recombinant vector synthesized the fusion protein and were also capable of removing the signal peptide from the original fusion protein, as judged by the assay of .betha.-lactamase activity and secretion into the growth medium by western blotting.

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Construction of Secretion Vectors Using the $\alpha$-amylase Signal Sequence of Bacillus subtilis NA64

  • Kim, Sung-Il;Lee, Se-Yong
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.74-81
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    • 1996
  • Two secretion vectors, pUBA240 and pUB340 were constructed by using the promoter and secretory signal region of the .alpha.-amylase gene from an .alpha.-amylase hyperproducing strain, Bacillus subtilis NA64. In this secretion vector system, various restriction enzyme sites are located immediately after the proregion of the .alpha.-amylase gene for easy replacement of various foregn structural genes. To evaluate this secretion vectors, the .betha.-lactamase gene of pBR322 was used as a reporter gene. The expressed and biologically active .betha.-lactamase was secreted into the culture broth from B. subtilis LKS86 transformants harboring each .betha.-lactamase secreting plasmid, pUBAbla and pUBSble. In both cases, more than 92% of expressed .betha.- lactamases were located idn the culture medium. The amount of the secreted .betha.-lactamase was about 80% of the total secreted proteins in the culture medium.

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Synthesis and ${\beta}-Lactamase$ Inhibitory Activity of 7-Exomethylene Cephalosporanates (7-엑소메칠렌 세팔로스포라네이트 유도체의 합성과 $\beta$- 락타메이즈 억제작용)

  • 이종민;최수항;이현수;임채욱;임철부
    • YAKHAK HOEJI
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    • v.43 no.6
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    • pp.782-788
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    • 1999
  • 7-Oxocephalosporanate 1 was treated with phosphonium salts 2~4 by Wittig reaction to afford 7-exomethylene cephalosporanates 5~7. They were oxidized to sulfones 8~10 with mCPBA. Deprotecton of benzhydryl 7-exomethylene cephalosporanate with $AlCl_3$ and NaHCO_3$ gave sodium salts of 7-exomethylene cephalosporanates 11~16. The ${\beta}-lactamase$ inhibitory activity of synthesized compounds 11~16 were compared with sulbactam, tazobactam and clavulanic acid against Type I, II, III, IV and TEM-2 $\beta$-lactamase in vitro. Compound 15 showed more potent activity than sulbactam and clavulanic acid against Type III, IV ${\beta}-lactamase$ enzyme.

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${\beta}-Lactamase$ Inhibitory Activity and Comparative Activity of 6-Benzothiazole Penicillin Derivatives in Combination with ${\beta}-Lactam$ Antibiotics (6-벤조치아졸 페니실린 유도체의 베타락타마제 효소억제력과 베타락탐항생제 병용시 활성비교)

  • Yoon, Sang-Bae;Im, Chae-Uk
    • YAKHAK HOEJI
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    • v.52 no.4
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    • pp.306-310
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    • 2008
  • In vitro ${\beta}-lactamase$ inhibitory activity of 6-benzothiazole penicillins (1, 2, 3 and 4) was compared with clavulanic acid, sulbactam and tazobactam. The inhibitory activity of exomethylene compounds (3 and 4) was stronger than those of non-exomethylene compounds (1 and 2). The sulfide 3 showed stronger inhibitory activity than sulbactam, clavulanic acid andsimilar to tazobactam against ${\beta}-lactamase$ Type I enzymes. The inhibitory activity of 4 was stronger than those of sulbactam, clavulanic acid and tazobactam against Type III and IV enzymes. The in vitro antimicrobial activity of ampicillin or cefoperazone combined with 3 or 4 was stronger than those of ampicillin or cefoperazone alone against many ${\beta}-lactamase$ producing strains to show that compounds 3 and 4 have some synergistic effect. The synergistic activity of 3 and 4 was comparable to sulbactam in some ${\beta}-lactamase$ producing strains, but it was inferior to tazobactam.

Survey of extended-spectrum β-Lactamase (ESBL) in pathogenic Escherichia coli isolated from poultry in Korea (국내 가금유래 병원성 대장균의 extended-spectrum β-lactamase(ESBL) 특성 조사)

  • Sung, Myung-Suk;Kim, Jin-Hyun;Cho, Jae-Keun;Seol, Sung-Yong;Kim, Ki-Seuk
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.259-265
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    • 2008
  • This study was conducted to investigate incidence of extended-spectrum ${\beta}$-lactamase (ESBL) producing strains and characteristics of ESBL gene in pathogenic Escherichia coli isolated from poultry during the period from April 2003 to December 2005 in Korea. Among 203 isolates, 4 isolates (3 from broilers and 1 from layer) were confirmed as ESBL producing strains by double disk synergy test, polymerase chain reaction and sequencing for ${\beta}$-lactamase genes. $bla_{CTX-M-15}$ and $bla_{CMY-2}$ were detected in these 4 isolates and were transferred to recipient by conjugation, respectively. Also, these ESBL producing strains were associated with multiple drug resistance. In conclusion, these results exhibit incidence of CTX-M and CMY-2 ${\beta}$-lactamase in pathogenic E coli from poultry in Korea, and clinically important meaning in human. And they also suggest the needs for rapid and broad surveillance to monitor ESBL genes and R plasmid transferring resistant gene in poultry.

Comparison of the Activities of Novel ${\beta}$-Lactamase Inhibitors, 6-Exomethylene Penamsulfones, with Other ${\beta}$-Lactamase Inhibitors as Combined with ${\beta}$-Lactam Antibiotics (I) (6위치 엑소 메칠렌 치환 페남계 베타락타마제 억제제의 베타락탐항생제와 병용시 활성비교(I))

  • Park, Kye-Whan;Kim, Ki-Ho;Kim, Mee-Young;Im, Chae-Uk;Yim, Chul-Bu
    • 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.

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Characterization of Plasmid-Mediated SHV-11 β-lactamase Gene of Klebsiella pneumoniae Isolated from the Clinical Specimens (임상검체로부터 분리한 플라스미드 매개성 SHV-11 β-lactamase 유전자의 특성)

  • Kim, Yun-Tae;Lee, Sang-Hoo;Jang, Ji-Hyun;Kim, Tae-Un;Choi, Seok-Cheol;Baik, Hyung-Suk;Lee, Kyoung-Ryul;Yoon, Hye-Ryoung;Kim, Young-Jin
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1718-1723
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    • 2009
  • In this study, we characterized extended-spectrum $\beta$-lactamase (ESBL)-producing Enterobacteriaceae isolated from clinical specimens in Korea and found two strains harboring plasmid-mediated $bla_{SHV-11}$, Klebsiella pneumoniae. First, the isolates were detected using the Vitek system and confirmed by the double-disk synergy test. The classification of gene coding for ESBL was also performed by polymerase chain reactions and followed by DNA sequencing. The transmission of genes was confirmed by transconjugation and transformation. Resistant expression of transformants was determined by broth microdilution minimal inhibitory concentration test. Genotypic analysis revealed that one strain harbored the $bla_{TEM-1}$, $bla_{SHV-11}$ and $bla_{CTX-M-15}$ and the other strain harbored the $bla_{SHV-11}$ and $bla_{CTX-M-15}$. They showed high resistance to oxyiminocephalosphorins (3rd-generation cephalosporins), while the transformant containing only $bla_{SHV-11}$ did not show any resistance to the antibiotics.

Characteristics and Antibiotic Susceptibility of Imipenem-Resistant Clinical Isolates Producing Carbapenemase (Carbapenemase를 생산하는 imipenem 내성 세균의 특성 및 항생제 감수성)

  • Choe, Han-Na;Park, Chul;Kim, Hyung-Rak;Baik, Keun-Sik;Kim, Se-Na;Seong, Chi-Nam
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1214-1220
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    • 2010
  • Imipenem-resistant bacteria were isolated from clinical specimens taken from hospitalized patients in Suncheon, Korea. Fifty-four isolates were phylogenetically analyzed based on 16S rRNA gene and gyrB gene sequence comparisons. Isolates were affiliated with Pseudomonas aeruginosa (30 strains; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2) and Pseudomonas putida (2). Twenty-two isolates produced metallo-$\beta$-lactamase (MBL); 12 Acinetobacter baumannii strains, 7 Pseudomonas aeruginosa strains, 2 P. putida strains and 1 Enterobacter hormaechei strain. Antibiotic susceptibility of the isolates was determined using the disc diffusion method and Vitek system. Strains producing metallo-$\beta$-lactamase (type IMP & VIM) were more resistant to antibiotics ceftazidime, aztreonam, amikacin and gentamicin than to strains producing OXA and SHV type of $\beta$-lactamase.