• Title/Summary/Keyword: Escherichia coli K12

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Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

Induction of Bactericidal Substance from the Immunized Larval Haemolymph of L. illustris (면역유충 체액으로 부터 항균물질의 유도)

  • 육순학;장정순
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.309-317
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    • 1987
  • It was known that normal-haemolymph from the 3rd instar larvae of Lucillia illustris contain a lysozyme (or lysozyme-like substance) with bactericidal activity to fram positive bacteria, and the bactericidal activity of injured-haemolymph was increased significantly after injuring the body wall. To elucidate the defence mechanism of insect against the nonpathogenic bacteria, the immune-haemolymph against Escherichia coli K-12 was prepared after immunization. The bactericidal activity between injured and immune-haemolymph was compared, and it was revealed that the immune-haemolymph showed higher titer of bactericidal activity to fram positive bacteria as well as to Escherichia coli. The bactericidal substance from the immune-haemolymph was purified through a successive chromatographies on Sephacryl S-300 and CM-Sepharose CL-6B, and it was characterized as a basic protein in nature with heat stable property at acidic conditions.

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Isolation and Culture Conditions of Hydrogen-producing Bacterium from Sewage Sludge (하수슬러지에서 수소생산 균주의 분리 및 배양조건)

  • Woo, Dae-Sik;Joo, Hyun;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.24 no.4
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    • pp.407-411
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    • 2013
  • Hydrogen producing bacterium, strain AS12 was isolated from the sludge of the anaerobic wastewater treatment process of south sewage treatment plant, Busan city. Phylogenetic analysis based on 16S rRNA sequence studies indicated that AS12 belonged to the genus Escherichia coli sp.. The optimum temperature and pH for hydrogen production were $35^{\circ}C$ and 8.0, respectively. The impact of the types and concentrations of carbon and nitrogen sources in the media on hydrogen production was investigated. The optimum carbon and nitrogen concentrations were 10 g/L of galactose and 5 g/L of peptone, respectively.

Antimicrobial Resistance and the Presence of Virulence Genes in Escherichia coli Strains Isolated from Ruditapes philippinarum in Gomso Bay, Korea (곰소만 해역의 바지락(Ruditapes philippinarum)에서 분리한 대장균 (Escherichia coli)의 항균제 내성 및 병원성 유전자의 보유성)

  • Kim, Tae-Ok;Eom, In-Seon;Park, Kwang-Ho;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.6
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    • pp.800-806
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    • 2016
  • In total, 151 Escherichia coli isolates from Ruditapes philippinarum in Gomso Bay were analyzed for their susceptibility to 18 different antimicrobial agents and for genes associated with virulence. For virulence genes, each strain of the isolates was positive for the enterotoxigenic E. coli (ETEC)-specific heat-stable toxin (estA), enteroinvasive E. coli (EIEC)-specific invasion-associated locus (iaa) gene and enteropathogenic E. coli (EPEC)-specific attaching and effacing (eae) gene. According to a disk diffusion susceptibility test, resistance to ampicillin was most prevalent (23.2%), followed by resistance to amoxicillin (22.5%), ticarcillin (20.5%), tetracycline (18.5%), nalidixic acid (12.6%), ciprofloxacin (10.6%), streptomycin (9.9%), and chloramphenicol (6.6%). More than 35.8% of the isolates were resistant to at least one antimicrobial agent, and 19.9% were resistant to four or more classes of antimicrobials; these were consequently defined as multidrug resistant. Minimum inhibitory concentration (MIC) ranges for the antimicrobial resistance of the 15 different antimicrobial agents of 54 E. coli strains were confirmed by varying the concentrations from $32-2,048{\mu}g/mL$. Overall, these results not only provide novel insights into the necessity for seawater and R. philippinarum sanitation in Gomso Bay but they also help to reduce the risk of contamination by antimicrobial-resistant bacteria.

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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Quantification of Inhibitory Impact of Heavy Metals on the Growth of Escherichia coli (중금속에 의한 대장균 성장 저해의 정량적 평가)

  • Jo, Gyeong-Suk;Gu, So-Yeon;Kim, Ji-Yeong;Ryu, Hui-Uk
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.341-346
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    • 2004
  • The quantification of the toxicological effects of the heavy metals such as Cu, Cd, Cr, Hg, and Zn on the growth of Escherichia coli was performed, and the variations of toxicwith exposure time were evaluated in adaptation procedure. The characteristics of growth inhibition on Escherichia coli by heavy metals were different with metal species, and critical concentration of each metal, which inhibited cell growth completely, were Cu of 3.5 mM, Zn of 2.5 mM, Cd of 1.5 mM, Cr of 1.2 mM, and Hg of 0.12 mM, respectively. The tolerance of E. coli against heavy metals, based on $EC_{50}$ values, increased in order of Cu > Zn > Cr > Cd > Hg. The slopes obtained from the relationship between ECso values and expose time corresponds to adaptability of test organisms to the toxicants. The adaptability of test organisms to the toxicants was much higher at higher slope values. Adaptability of E. coli on heavy metals increased in order to Zn > Cd > CU >> Cr> Hg.

Antimicrobial Resistance of Escherichia coli Isolates from Mussel Mytilus galloprovincialis Farms and Inland Pollution Sources in the Changseon Area, Korea (창선 해역의 지중해담치(Mytilus galloprovincialis) 양식장 및 육상오염원에서 분리한 대장균(Escherichia coli)의 항생제 내성)

  • Kwon, Soon Jae;Lee, Ka Jeong;Jung, Yeoun Joong;Park, Sang Gi;Go, Kyeong Ri;Yang, Ji Hye;Mok, Jong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.564-572
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    • 2016
  • In this study, we isolated and characterized Escherichia coli from mussels and inland pollution sources in or in proximity to the Changseon area on the southern coast of Korea in 2014. A total of 147 strains of E. coli were isolated from 54 mussels and 32 pollution-source samples. The susceptibility of the isolates to 24 antimicrobial agents was analyzed. The resistance of E. coli isolates to rifampin was highest at 100%, followed by cephalothin (98.6%), tetracycline (91.8%), amikacin (81.0%), ampicillin (79.6%), cefazolin (79.6%), streptomycin (73.5%), piperacillin (70.7%), gentamicin (37.4%), cefoxitin (35.4%), cefamandole (34.7%), tobramycin (29.9%), amoxicillin/clavulanic acid (24.5%), nalidixic acid (21.8%), trimethoprim (19.0%), chloramphenicol (17.7%), cefotaxime (12.9%), trimethoprim (10.9%), ceftazidime (10.2%), aztreonam (7.5%), imipenem (2.7%), cefepime (2.0%), and cefotetan (0.0%). In addition, the antimicrobial resistance of E. coli isolates from inland pollution sources was slightly greater than or similar to that of isolates from mussels.

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.

Interaction for Phosphotransfer between N-Terminal Domain of Enzyme I and HPr of E. coli Phosphoenolpyruvate:Sugar Phosphotransferase System.

  • Seok, Yeong-Jae
    • Proceedings of the Korean Biophysical Society Conference
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    • 1998.06a
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    • pp.12-12
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    • 1998
  • The interaction between the N-terminal domain of enzyme I (EIN) and the histidine-containing phosphocarrier protein HPr of the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system has been investigated by Isothermal Titration Calorimetry and heteronuclear magnetic resonance spectroscopy.(omitted)

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Genotype by Dietary Lysine Interaction for Growth and Response to Sheep Red Blood Cells and Escherichia coli Inoculation in Commercial Broiler Chicks

  • Praharaj, N.K.;Reddy, M.R.;Panda, A.K.;Rama Rao, S.V.;Sharma, R.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.8
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    • pp.1170-1177
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    • 2002
  • An experiment was conducted to assess the interaction between genotypes and dietary lysine content in commercial broiler chicks by measuring growth, and response to sheep red blood cells (SRBC) and Escherichia coli (E.coli) inoculation. Female chicks from four genotypes (A=Anak 2000; B=Hubbard; C=Cobb and D=Synthetic broiler) were fed four levels of lysine in diet from d old till the end of experiment. The lysine content of the diet was 9.61, 10.51, 11.41 and 12.31 g/kg. Body weights at 0, 14, 28 and 42 d of age and pen-wise feed intake till 14, 28 and 42 d of age were recorded. Production of antibody against SRBC and resistance to E.coli were measured at 5 d of post inoculation (PI) at 43 d of age. Also, response to phytohemaglutinin-P (PHA-P) was measured at 12 and 24 h of PI at 48 d of age. Genotype by dietary lysine interaction was significant for body weights at 14 and 28 d of age, but not at 42 d of age. Genotype by dietary lysine interaction was not significant for feed efficiency, for antibody titers against SRBC, and for air sac lesion score, relative bodyweight change, and relative weights of bursa and spleen in response to E.coli inoculation. However, a significant interaction was observed between the levels of lysine and dosage of SRBC for antibody titers. There was significant genotype by dietary lysine interaction for cutaneous basophilic hypersensitivity (CBH) response to PHA-P at 12 and 24 h of PI. It may be concluded that to obtain optimum body weight and immunity in commercial broilers the dietary lysine requirement may be recommended specific to the genotype.