• Journal of Microbiology and Biotechnology
• /
• v.17 no.3
• /
• pp.474-480
• /
• 2007

Fusarium wilt of cucumbers was effectively controlled by Escherichia coli expressing an endochitinase gene (chiA), and the rate was as effective (60.0%) as the wild-type strain S. proteamaculans 3095 (55.0%) where the gene was cloned. However, live cells of soil inoculated E. coli host harboring the chiA gene did not proliferate but declined 100-fold from $10^8$ CFU during the first week and showed less than 10 cells after day 14, suggesting that E. coli was able to express and produce the chitinase enzyme to the soil even as the population was gradually decreasing. Because the majority of the strains was alive for only a short period of time and the Fusarium-affected seedlings showed symptoms of wilting within 7-10 days, it seems that the pathogen control was decided early after the introduction of the biocontrol agent, eliminating the survival of the antagonist. These results indicated that soil inoculated E. coli could sufficiently express and produce the recombinant protein to control the pathogen, and root or soil colonization of the antagonist might not be a significant factor in determining the efficacy of biological control.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.6
• /
• pp.765-770
• /
• 2014

This study aimed to characterize CTX-M producers of urinary E. coli and K. pneumoniae isolates and to determine the prevalence of plasmid-mediated antimicrobial resistance genes among them. Minimum inhibitory concentrations (MICs) were determined, and PCR and sequencing were performed. Among the 42 (82.3%) E. coli and 24 (77.4%) K. pneumoniae isolates containing $bla_{CTX-M}$, $bla_{CTX-M-14}$ and $bla_{CTX-M-15}$ were detected in 23 and 19 E. coli isolates, respectively, and in 7 and 17 K. pneumoniae isolates, respectively. CTX-M producers of urinary E. coli and K. pneumoniae were resistant to multiple antibiotics and contained other antimicrobial resistance genes. CTX-M-15 producers contained more antimicrobial resistance genes than did CTX-M-14 producers.

• Fisheries and Aquatic Sciences
• /
• v.14 no.3
• /
• pp.226-229
• /
• 2011

This study was conducted to develop a proficiency test (PT) specimen for the enumeration of Escherichia coli, a sanitary indicative bacterium, in molluscan bivalve shellfish. The common mussel Mytilus edulis was chosen as a matrix for the PT specimen. Mussels were artificially bioaccumulated for approximately 3 h with E. coli culture. After determining the homogeneity of samples, samples were distributed to 17 participants involved in the proficiency testing program. The enumeration of E. coli was performed by the most probable number method of the American Public Health Association. Statistical evaluations of the results obtained from inter- and intra-laboratory variation indicated no significant differences in the accuracy of these techniques between participants, indicating z-scores of ${\leq}{\pm}2$ and suggesting that preparation of the PT specimen for enumerating E coli in shellfish was successful.

• Microbiology and Biotechnology Letters
• /
• v.23 no.5
• /
• pp.550-555
• /
• 1995

A strain of Pseudomonas sp. isolated from soil was shown to produce a high level of extracellular endo-inulinase. In this work, the endo-inulinase gene (inu1) of the bacterial strain was cloned into the plasmid pBR322 by using EcoRI restriction endonuclease and E. coli HB101 as a host strain. One out of 7, 000 transformants obtained from the above cloning experiment formed a clear zone around its colony on the selective medium supplemented with 2.0% inulin after a prolonged incubation at 37$\circ$C and subsequent cold shock treatment. The functional clone was found to carry a recombinant plasmid (pKMG50) with a 3.7 kb genomic insert containing the genetic information for the inulinase activity. The inulinase from E. coli HB101/pKMG50 was proved to be an endo-acting enzyme and produced constitutively in the recombinant E. coli cells. Zymogram of the enzyme from the recombinant cells with inulin substrate indicated that the molecular mass of the active protein was 190 Kd, while that of the endo-inulinase from the Pseudomonas strain was 170 Kd. This size discrepancy suggested that the inulinase from the recombinant E. coli HB101 cells might be the initial product of translation, not the mature form produced in the strain of Pseudomonas sp..

• Journal of Microbiology and Biotechnology
• /
• v.30 no.10
• /
• pp.1552-1558
• /
• 2020

With an increase in the consumption of non-heated fresh food, foodborne shiga toxin-producing Escherichia coli (STEC) has emerged as one of the most problematic pathogens worldwide. Endolysin, a bacteriophage-derived lysis protein, is able to lyse the target bacteria without any special resistance, and thus has been garnering interest as a powerful antimicrobial agent. In this study, rV5-like phage endolysin targeting E. coli O157:H7, named as LysECP26, was identified and purified. This endolysin had a lysozyme-like catalytic domain, but differed markedly from the sequence of lambda phage endolysin. LysECP26 exhibited strong activity with a broad lytic spectrum against various gram-negative strains (29/29) and was relatively stable at a broad temperature range (4℃-55℃). The optimum temperature and pH ranges of LysECP26 were identified at 37℃-42℃ and pH 7-8, respectively. NaCl supplementation did not affect the lytic activity. Although LysECP26 was limited in that it could not pass the outer membrane, E. coli O157: H7 could be effectively controlled by adding ethylenediaminetetraacetic acid (EDTA) and citric acid (1.44 and 1.14 log CFU/ml) within 30 min. Therefore, LysECP26 may serve as an effective biocontrol agent for gram-negative pathogens, including E. coli O157:H7.

• Environmental Engineering Research
• /
• v.21 no.2
• /
• pp.211-218
• /
• 2016

Cheonggye-cheon is a partly restored urban stream located in central Seoul. We monitored fecal contamination using three different fecal indicators, total coliforms (TC), fecal coliforms (FC) and E. coli, to assess differences in each indicator on days of varying weather conditions. Presumptive TC, FC and E. coli colonies were identified by their 16S rRNA sequences. The results showed that enumeration of E. coli provided a better reflection of fecal contamination of the stream than TC and FC. The main sources of contamination were the inflow of fecal-polluted groundwater from the vicinity of a subway line and two inflowing streams. The fecal contamination was worsened on days with heavy rain because untreated sewage from a collecting facility flowed into the stream. Moreover, growth potential of fecal indicator (E. coli) in situ induced by algal exudates was measured. Our results suggest that an appropriate standard based on E. coli rather than TC and FC should be established for improving water quality management strategies of Cheonggye-cheon in the future.

• YAKHAK HOEJI
• /
• v.41 no.3
• /
• pp.359-364
• /
• 1997

Resistance gene mphK was cloned from Escherichia coli 209K strain which is highly resistant to erythromycin (EM). By using the cloned plasmid pGE64, E. coli NM522 was transformed. The comparison of macrolide-phosphotransferase K [MPH(K)] activity between E. coli 209K and E. coli NM522(pGE64) showed that the total enzyme activity of MN522(pGE64) was fifty-fole higher than that of 209K. To identify characteristics of MPH(K) more precisely. MPH(K) was isolated and purified from the NM522 (pGE64). The final purification f MPH(K) through several stages of purification process was 89 fole and the overall recovery was 11%. This enzyme was monomer with the molecular weight of 34 kDa and its isoelectric point (pI) was 5.0. The optimal pH and temperature for activity were 8.0 and $40^{\circ}C$, respectively.

• Korean Journal of Veterinary Research
• /
• v.30 no.3
• /
• pp.287-295
• /
• 1990

The purpose of this study was the examination for presence of pilus antigen, O serogroups, colicin production, antibiotic susceptibility and plasmid profiles among E coli isolated from diarrheal piglets and fattening pigs in Taegu province. Of 145 E coli isolated, 98 strains (67.4%) possesed pilus antigens which belonged to either K88 (47.6%), K99 (11.7%) or 987P (8.3%) types. Fifty-nine strains (40.7%) were classified into tenO serogroups and their types were O8 (22.0%), O20(16.9%), O141(15.3%), O9(10.2%), O45(10.2%), O139(8.5%), O064(6.8%), O149(5.0%), O157(3.4%), and O115(1.7%). Thirty-three strains (22.8%) were colicinogenic and 6 strains (4.1%) were hemolytic. One hundred and thirty-nine strains (95.9%) of 145 E coli isolates were resistant to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, rifampicin and nalidixic acid, alone or in combination thereof. Ninety strains (64.7%) of 139 drug resistant strains carried R factor (R) which were transferable to the recipient by conjugation. In gel electrophoresis for the isolation of plasmid DNA, the number of plasmid DNA band varied from 2 to 11 in 16 E coli with pilus antigen. It's molecular weight ranged from 1.0 to 60.0 megadalton.

• YAKHAK HOEJI
• /
• v.35 no.2
• /
• pp.61-72
• /
• 1991

These studies were made to assess the present stage of resistance to antibiotics, incidence and transferability of R-factors against E. coli. From March to July 1987, 59 strains of E. coli were isolated from specimens of patients collected at university hospitals in Seoul, 64 strains from stools of domestic animals and 66 strains from drainages in Seoul. These specimens were tested for resistance to 12 kinds of antimicrobial agents by means of agar dilution method. Using Muller-Hinton agar for the assay of drug resistance and tryptic soy broth as propagating medium for conjugation. The strains of E. coli were found to be resistant to one or more antibiotics and were considered to be potential donors of R-plasmid. The resistant strains of E. coli isolated from patients, domestic animals and drainages were found to be 55(93%), 33(52%) and 31(47%), respectively. Resistance to Tc, Ap and Cb was the highest in those isolated from patients and drainages, and resistance to Tc, Cm and Sm was the highest in those isolated from domestic animals. In the transfer test of drug resistance by conjugation method, 17 strains (47%) isolated from patients, 15(54%) isolated from domestic animals and 15(56%) isolated from drainages showed positive results, transperable resistant plasmid molecules with variable range in each strain.

• Molecules and Cells
• /
• v.26 no.6
• /
• pp.616-620
• /
• 2008

Maintaining redox balance is one of the crucial requirements for a cell to endure stress from the outside. Dehydroascorbate reductase (DHAR; EC 1.8.5.1) plays an important role in the ascorbate-glutathione cycle; one of the major ROS scavenging systems in most known biological systems. A cDNA clone of the DHAR gene from Oryza sativa (OsDHAR) was isolated and overexpressed in Escherichia coli BL21 (DE3) strain from the pET-28a(+) expression vector. The OsDHAR transformed E. coli cells showed significantly higher DHAR activity and a lower level of ROS than the E. coli cells transformed by an empty pET-28a(+) vector. Also, the DHAR-overexpressing E. coli strain was more tolerant to oxidant- and heavy metal-mediated stress conditions than the control E. coli strain. The results suggest that the overexpressed rice DHAR gene effectively functions in a prokaryotic system and provide protection to various oxidative stresses.