• Title/Summary/Keyword: E. coli K-12

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Microbial Risk Assessment of Non-Enterohemorrhagic Escherichia coli in Natural and Processed Cheeses in Korea

  • Kim, Kyungmi;Lee, Heeyoung;Lee, Soomin;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.37 no.4
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    • pp.579-592
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    • 2017
  • This study assessed the quantitative microbial risk of non-enterohemorrhagic Escherichia coli (EHEC). For hazard identification, hazards of non-EHEC E. coli in natural and processed cheeses were identified by research papers. Regarding exposure assessment, non-EHEC E. coli cell counts in cheese were enumerated, and the developed predictive models were used to describe the fates of non-EHEC E. coli strains in cheese during distribution and storage. In addition, data on the amounts and frequency of cheese consumption were collected from the research report of the Ministry of Food and Drug Safety. For hazard characterization, a doseresponse model for non-EHEC E. coli was used. Using the collected data, simulation models were constructed, using software @RISK to calculate the risk of illness per person per day. Non-EHEC E. coli cells in natural- (n=90) and processed-cheese samples (n=308) from factories and markets were not detected. Thus, we estimated the initial levels of contamination by Uniform distribution ${\times}$ Beta distribution, and the levels were -2.35 and -2.73 Log CFU/g for natural and processed cheese, respectively. The proposed predictive models described properly the fates of non-EHEC E. coli during distribution and storage of cheese. For hazard characterization, we used the Beta-Poisson model (${\alpha}=2.21{\times}10^{-1}$, $N_{50}=6.85{\times}10^7$). The results of risk characterization for non-EHEC E. coli in natural and processed cheese were $1.36{\times}10^{-7}$ and $2.12{\times}10^{-10}$ (the mean probability of illness per person per day), respectively. These results indicate that the risk of non-EHEC E. coli foodborne illness can be considered low in present conditions.

Purification and Characterization of the Overproduced E. coli Endochitinase (과량 생산된 대장균 chitin 분해효소의 정제 및 특성 조사)

  • Hwang, Hee-Young;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.171-175
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    • 2003
  • The putative endochitinase gene, yheB of Escherichia coli K-12 is not expressed under lab culture conditions. The endochitinase gene was amplified by PCR and subcloned into pET28c vector and pQE9 vector, respectively. The endochitinase produced in E. coli harboring pET28c containing yheB or pQE9 vector containing yheE was partly released into the growth medium. The overproduced endochitinase was partially purified by His affinity column chromatography and DE-52 column chromatography. The apparent molecular weight of the endochitinase determined by SDS-polyacrylamide gel electrophoresis was about 97,000. The purified E. coli endochitinase showed maximal chitinolytic activity at pH 6 and $40^{\circ}C$.

Construction of Plasmids for Overproduction of L-Phenylalanine (L-페닐알라닌 대량생산을 위한 재조합 플라스미드 구성)

  • Lee, Sae-Bae;Park, Chung;Won, Chan-Hee;Choi, Duk-Ho;Lim, Bun-San
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.169-173
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    • 1990
  • For the overproduction of L-phenylalanine using Escherichia coli, the authors constructed various recombinant plasmids including pMW 10, pMW 11 and pMW 12. The $aroF{FR}$ and $pheA^{FR}$ genes for the production of L-phenylalanine were isolated from Escherichia coli MWEC 101-5 strains. The productivity and atability of Escherichia coli regulatory mutants containing recombinant plasmids were investigated to evaluate the efficiency of the $aroF^{FR}$ and $pheA^{FR}$ genes. The MWEC 101-5/pMW 11 strain produced 24.3g/l of L-phenylalanine while its stability was 73.8 percent. The specific activity of prephenate dehydratase in the MWEC 101-5/pMW 11 strain increased by 26-fold compared with that of Escherichia coli K-12.

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Typing of Extended-Spectrum ${\beta}-Lactamase$ of Escherichia coli and Klebsiella pneumoniae Isolated from Rivers in Busan, Korea (부산지역 하천에서 분리된 장내세균 Escherichia coli와 Klebsiella pneumoniae의 광범위 베타 락탐 분해효소 (Extended-Spectrum ${\beta}-Lactamase$)에 대한 유형별 분류)

  • Lee, Hun-Ku;Kim, Hye-Jin;Kim, Gun-Do
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.116-123
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    • 2007
  • The purpose of this study was typing the plasmid mediated extended spectrum ${\beta}-lactamases$ produced by enteric bacteria isolated from rivers in Pusan. Six strains of Eschericha coli and fifteen strains of Klebsiella pneumoniae transferred their plasmid mediated extended spectrum ${\beta}-lactamase$ genes to the recipient strain Eschericha coli J53 $Azid^{R}$. The plasmid mediated extended spectrum ${\beta}-lactamase$ genes were sequenced directly after PCR and the types were determined by the BCM Search Launcher and GenBank nucleotid database. Determined types of the plasmid mediated extended spectrum ${\beta}-lactamases$ were TEM-52 and SHV-12. TEM-52 was isolated from both Escherichia coli and Klebsiella pneumoniae. However SHV-12 was isolated from Klebsiella pneumoniae only. The results indicated that the plasmid mediated extended spectrum ${\beta}-lactamase$ producing bacteria spreded over the area of clinical to the nature in Korea.

Synergistic Inhibition of Escherichia coli by a Combination of Bacteriophage and Organic Acid (박테리오파지와 유기산의 병용처리에 의한 효율적인 대장균 생육 억제)

  • Kim, Seon-Gyu;Moon, Gi-Seong
    • Journal of Food Hygiene and Safety
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    • v.34 no.6
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    • pp.591-594
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    • 2019
  • Some strains of Escherichia coli are categorized as pathogenic bacteria and alternative antimicrobials including bacteriophages for controlling these bacteria have been studied. In this study we screened antimicrobial candidates that present synergistic inhibition of the growth of E. coli DH5α as a model when co-treated with the bacteriophage ECP27 to target the bacteria. As candidates, CaCl2, lactic acid, and citric acid were tested. CaCl2 showed a synergistic inhibition against the strain by dose-dependent manner at 6 h of incubation but the viable cell count was recovered at 12 h. However, lactic acid and citric acid at 30 mM concentration showed synergistic inhibitions at 6 h of incubation and cleared the viable cells of E. coli DH5α at 12 h when co-treated with the bacteriophage even though lactic acid or citric acid alone was effective. Therefore, co-treatment using the bacteriophage and organic acids such as lactic acid and citric acid can be a solution for synergistic inhibition of the growth of E. coli.

The DNA region of rtn gene essential for resistance against N4 infection (N4에 대해 내성을 나타내는데 필요한 rtn 유전자의 부위)

  • 이동환;유선미;황의욱;이영훈;채건상
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.290-295
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    • 1991
  • N4 phage, which infects E. coli K-12 strains, could not infect E. coli K-12 strains containing rtn(resistant to N4) gene on plasmids, which was isolated from Proteus vulgaris ATCC 13315. The region of rtn gene for Rtn phenotype was reduced to the 1.7 kb HincII-AccI fragment, and rtn gene seemed to have its own promoter. This putative promoter was present in 107 bp HindII-DraI fragment, and known to be functional in E. cole K-12, which is supported by the fact that phenotype of a subclone, pRMG103A1B which does not contain the 107 bp fragment, was dependent on the existance of a functional promoter in the upstream of rtn gene, and that the 107 bp fragment had promoter activity when located in the upstream of structural gene of galactodinase of E. coli. The promoter-bearing fragment contains two overlapping putative promoter sequences, both of which show a fit in eight of twelve nucleotides with consensus sequences of E. coli promoters at the -35 and -10 regions.

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Studies on the Antibiotic Sensitivity of Escherichia coli Isolated from a Doctorless Area and Seoul National University Hospital (일부(一部) 무의촌지역(無醫村地域)과 병원(病院)에서 분리(分離)한 E.coli의 항생제(抗生劑) 감수성(感受性)에 대(對)하여)

  • Kim, Ik-Sang;Shin, Hee-Sup;Rhee, Kwang-Ho;Cha, Chang-Yong;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.12 no.1
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    • pp.1-10
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    • 1977
  • 40 strains of E. coli isolated from residents of a doctorless area in Korea in 1976 and 40 strains of E. coli isolated from patients of Seoul National University Hospital from 1975 to 1976 were examined for susceptibilities to 14 antimicrobial agents by the agar dilution method. The susceptibilities of the two groups to each antimicrobial agent were compared and correlations in the antimicrobial susceptibility of the 80 strains of E. coli among the 14 antimicrobial agents were also analyzed. The results were obtained as follow: 1. With Tetracycline, Oxytetracycline, Doxycycline and Ampicillin, the mean MIC's of E. coli isolated from patients of Seoul National University Hospital were 8.6 to 14 times higher than. those of E. coli isolated from residents of a doctorless area. 2. With Streptomycin, Minocycline and Carbenicillin, the mean MIC's o{ E. coli isolated from patients of Seoul National University Hospital were 4.1 to 5.6 times higher than those of E. coil isolated from residents of a doctorless area. 3. With Kanamycin, Penicillin and Cotrimoxazole, the mean MIC's of E. coli isolated from patients of Seoul National University Hospital were 2.6 to 3.7 times higher than those of E. coli isolated from residents of a doctorless area. 4. There were no significant differences in susceptibility to Erythromycin respectively between E. coli isolated from patients of Seoul National University coli isolated from residents of a doctorless area. 5. E. coli isolated from patients of Seoul National University Hospital were resistant to Erythromycin(100%), Streptomycin(75%), Tetracycline(72.5%), Oxytetracycline(72.5%), Doxycycline(72.5%), Minocycline(67.5%), Penicillin(82.5%), Ampicillin(60%) and Carbenicillin(65%) respectively and were sensitive to Gentamicin(97.5%), Cephalexin(92.5%) and Kanamycin(72.5%) respectively. 6. E. coli isolated from residents of a doctorless, area were resistant to Erythromycin(100%), Streptomycin(40%) and Penicillin(50%) respectively and were sensitive to Gentamicin(100%), Kanamycin(92.5%), Tetracycline(87.5%), Oxytetracycline(87.5%), Doxycycline(87.5%), Minocycline(87.5%), Ampicillin(95%), Carbenicillin(92.5%) and Cephalexin(97.5%) respectively. 7. There were high correlations among the suscebtibilities of the 80 strains of E. coli to Tetracycline analogues(Tetracycline, Oxytetracycline, Doxycycline and Minocycline) and among susceptibilities of the 80 strains of E. coli to Penicillin analogues(Penicillin, Ampicillin and Carbenicillin). 8. There were relatively high correlations between the susceptibilities of the 80 strains of E. coli to Penicillin analogues and those to Tetracycline analogues, between the susceptibilities to Penicillin analogues and those to Streptomycin and between the susceptibilities to Tetracycline analogues and those to Streptomycin.

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Functional Analysis of the marB gene of Escherichia coli K-12

  • Lee, Chang-Mi;Park, Byung-Tae
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.153-161
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    • 2004
  • Antibiotic resistance is often associated with the production of inner membrane proteins (for example, AcrAB/TolC efflux pump) that are capable to extrude antibiotics, detergents, dyes and organic solvents. In order to evaluate the unknown MarB function of Escherichia coli, especially focused on the function of OmpF porin, several mutants were construted by T4GT7 transduction. MarA plays a major roles in mar (multiple antibiotic resistance) phenotype with AcrAB/TolC efflux pump in E. coli K-12. Futhermore, MarA decreases OmpF porin expression via micF antisense RNA. Expression of acrAB is increased in strains containing mutation in marR, and in those carrying multicopy plasmid expressing marA. MarB protein of E. coli K-12 showed its activity at OmpF porin & TolC protein as target molecule. Some paper reported MarB positively regulates OmpF function. MarA shows mar phenotype, and MarB along with MarA show decreased MIC through OmpF function. By this experiment, MarB could decrease MIC through the OmpF porin & TolC protein as target.

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Development and Validation of Predictive Models of Esherichia coli O157:H7 Growth in Paprika (파프리카에서 병원성 대장균의 성장예측 모델 개발 및 검증)

  • Yun, Hyejeong;Kim, Juhui;Park, Kyeonghun;Ryu, Kyoung-Yul;Kim, Byung Seok
    • Journal of Food Hygiene and Safety
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    • v.28 no.2
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    • pp.168-173
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    • 2013
  • This study was carried out to develop and validate predictive models of E. coli O157:H7 growth. Growth data of E. coli O157:H7 in Paprika were collected at 12, 24, 30 and $36^{\circ}C$. The population increased into 3.0 to 3.8 log10 CFU/g within 4 days, then continued to increase at a slower rate through 10 days of storage at $12^{\circ}C$. The lag time (LT) and maximum specific growth rate (SGR) obtained from each primary model was then modeled as a function of temperature using Davey and square root equations, respectively. For interpolation of performance evaluation, growth data for a mixture of E. coli O157:H7 were collected at time intervals in paprika incubated at the different temperatures, which was not used in model development. Results of model performance for interpolation data demonstrated that induced secondary models showed acceptable goodness of fit. Relative errors in the LT and SGR model for interpolation data (18 and $27^{\circ}C$) was 100%, which show acceptable goodness of fit and validated for interpolation. The primary and secondary models developed in this study can be used to establish tertiary models to quantify the effects of temperature on the growth of E. coli O157:H7 in paprika.

A Study on The Effect of Ginseng Saponin Fraction on Cell Wall (인삼 사포닌 분획이 세포벽에 미치는 영향)

  • Jo, Yeong-Dong;Kim, Tae-U;Choe, Hae-Gil
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.65-72
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    • 1981
  • In this experiment, observations were made on the effects of ginseng saponin, one of the major components of Korean ginseng (Panax ginseng, C. A. Meyer) root, on the membranes of microorganism (E. coli K-12), the concentration of intracelluar and extracellular cycle AMP therein, and uptake of U-14C-glucose. When the E. coli were grown on media containing 0.1% ginseng saponin, the growth was faster than for that of the control by about 30 minutes. The lysis of E. coli grown on the ginseng saponin medium increased to about double that of the control in the stationary phase. And the amount of protein and lipopolysaccharides in the outer cell meberances increased 25% and 80% respectively in comparison with the control. By electron microscope observation, it was shown that the periplasmic region of the E. coli grown on the ginseng saponin medium was widened it was observed that the cellular cyclic AMP content of the E. coli increased significantly to the hightest levels between the late exponential phase and early stationary phase. The total cyclic AMP content of E. coli grown on the ginseng saponin medium decreased about 50% when compared to that of the control.

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