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Exprission of cellulomonas biazotea cellobiase gene in E. coli (Cellulomonas biazotea cellobiase gene의 대장균에의 형질발현)

  • 박영길;연창석;최영길
    • Korean Journal of Microbiology
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    • v.26 no.1
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    • pp.6-12
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    • 1988
  • Cellobiase ($\beta$-glucosidase) is an enzyme of the cellulase system in cellulolytic microor-ganisms. The chromosomal DNA fragment which include cellobiase gene of Cellulomonas biazotea was cloned in Eschericia coli via plasmid pBR 322 vector. Restriction enzyme Sal I was used to obtain adequate size of fragments from C. biazotea. chromosomal DNA. The transformant of E. coli HB101 with recombinant plasmid pBG101 showed cellobiase activity, which is not ordinary in E. coli HB101. The enzyme activity of the transformant was as of 20% lower than that of C. biazotea.

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CYTOTOXICITY OF PATULIN AND ITS EFFECT ON THE LAMBDA DNA CLEAVAGE BY RESTRICTION ENDONUCLEASE

  • Lee, Kil-Soo
    • Toxicological Research
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    • v.7 no.2
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    • pp.157-163
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    • 1991
  • The effect of patulin, a mycotoxin, on the growth of Escherichia coli cell was investigated. E. coli cell elongation usually shown in SOS-response for DNA repair was induced by 20 mg of patulin per ml. After staining the E. coli chromosome with fluorescence dye(DAPI, 4', 6-diamino-2-phenyl-indole), chromosomal DNA partitioning was not affected by patulin. The observation indicateds that patulin acts as a DNA damaging agent which is effective for E. coli cell elongation introduced by the inhibition of septum formation.

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Expression and Characterization of Escherichia coli Adhesin Protein Linked to Cholera Toxin A2/B Subunits in Escherichia coli

  • Lee, Young-Hwa;Ryu, Dong-Kyun;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.309.2-309.2
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    • 2002
  • The FimH subunit of type l-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically coupled to the ctxa2b gene, which was then cloned into pMAL -p2E expression vector. The chimaeric construction of pMALfimH/ctxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

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Effect of Oregano (Origanum vulgare L.) on the Survival of Escherichia coli ) O157:H7 and Staphylococcus aureus 196E during Cold Storage (저온저장중 Oregano(Origanum vulgare L.)가 Eschetichia coli O157:H7과 Staphylococcus aureus 196E의 생존에 미치는 영향)

  • 박찬성;박금순;김미림
    • Korean journal of food and cookery science
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    • v.13 no.4
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    • pp.440-447
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    • 1997
  • The antibacterial effect of low concentrations of oregano (Origanum vulgare L.) in culture broth against Escherichia coli O157:H7 and Staphylococcus aureus 196E was tested at 35,5 and -20$^{\circ}C$. Tryptic soy broth (TSB) containing 0∼2% (w/v) of oregano was inoculated with 10$\^$6/∼10$\^$7/ CFU/$m\ell$ of E. coli or S. aureus and incubated at each temperature. The growth of E. coli was not inhibited at 0.1∼1.0% oregano and the growth occured at 2% oregano but only after a prolonged lag period. The death rate of E. coli after stationary phase was increased with increasing concentration of oregano in culture broth. The growth of S. aureus was inhibited with increasing concentration of oregano at 35$^{\circ}C$. Growth of S. aureus occured at the presence of 0.3∼0.5% oregano after a long lag period while the viability at 1.0∼2.0% was decreased during storage at 35$^{\circ}C$. During refrigerated storage at 5$^{\circ}C$, inhibition of E. coli or S. aureus was increased with the progress of time and increasing spice concentration. At the presence of 0.5∼2.0% oregano, E. coli and S. aureus were dead after 20 and 16 days of refrigerated storage, respectively. During frozen storage at -20$^{\circ}C$, the antibacterial activity of oregano against E. coli was increased with increasing storage time and spice concentration while the antibacterial activity against S. aureus was effective during the early period of storage, and no changes in the population of S. aureus occurred at different concentrations of oregano during frozen storage. Viable counts of E. coli were 1/3∼l/7 and S. aureus were 1/18∼l/46 of the control at 0.1% oregano in culture broth during frozen storage.

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Efficient Treatment Methods for Reducing Escherichia coli Populations in Commercially-Available Red Pepper Powder in Korea (국내 유통 고춧가루의 병원성 대장균 오염 및 대장균 저감화 방법)

  • Song, Young-Jin;Park, Se-Won;Chun, Se-Chul;Choi, Mi-Jung;Chung, Koo-Chun;Lee, Si-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.6
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    • pp.875-880
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    • 2012
  • This study was conducted to investigate the level of contamination of pathogenic Escherichia (E.) coli in 50 types of red pepper powders collected domestically. Pathogenic E. coli was confirmed using real-time PCR to confirm the 4 types of EAEC, EPEC, EHEC and ETEC. One sample out of 50 was contaminated with pathogenic E. coli. The type of pathogenic E. coli detected in the sample was EAEC. This study was also conducted to determine the effect of alcohol treatment on the reduction of E. coli populations in red pepper powder. The amount of E. coli in the control was $1.2{\times}10^6$ cfu/mL. The amount of E. coli in 10 minutes immersion treatment with 10% alcohol was $1.1{\times}10^6$ cfu/mL. In samples treated with over 20% alcohol, E. coli was not detected. This showed that 10 minutes of immersion in over 20% alcohol might be effective to reduce E. coli. This study was also conducted to determine the effect of UV irradiation on E. coli reduction. The number of E. coli in the control group was $5.0{\times}10^5$ cfu/mL. However, the number of E. coli in 45 min of the UV irradiated sample decreased to $1.0{\times}10^3$ cfu/mL, by $10^2$ cfu/mL. In contrast, E. coli was not detected in an over 60 min UV irradiated sample in $10^{-2}dilution$. This study showed that over 20% alcohol treatment and UV irradiation for 60 min was effective to control E. coli in red pepper powder.

Antimicrobial Activity of Sodium chlorate and Korean Herbal Extracts against Mice infected with Escherichia coli O157:H7 (Escherichia coli O157:H7에 감염된 마우스에 대한 염소산나트륨과 한약재 복합추출물 합제의 항균효과)

  • Cha, Chun-Nam;Lee, Yeo-Eun;Son, Song-Ee;Park, Eun-Kee;Choi, Hyun-Ju;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.81-86
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    • 2012
  • The present study was evaluated the antibacterial effect of the combination of $Coptidis$ $rhizoma$, $Glycyrrhiza$ $uralensis$ Fischet, $Schizandra$ $chinensis$ and $Corni$ $Fructus$(1:1:1) extracts(CGSC10). Furthermore, the effectiveness of CGSC10, sodium chlorate, and the combination of CGSC10 and sodium chlorate(CGSCS10) against $E.$ $coli$ O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 5, 10, and 20% CGSC10 was inhibited the growth of $E.$ $coli$ O157:H7 by 34.7, 60.2, and 76.4%, respectively. For 7 days after single challenge with $E.$ $coli$ O157:H7, forty female ICR mice were divided into four experimental groups which were administered in drinking water with saline, 10% CGSC10, 15 mM sodium chlorate, and CGSCS10, respectively. On the 3rd day, the number of $E.$ $coli$ O157:H7 in mouse feces was significantly decreased by administration of CGSC10, 15 mM sodium chlorate, and CGSCS10 ($p$ < 0.001). On the 7th day-after administration, CGSC10, sodium chlorate, and CGSCS10 were decreased the number of $E.$ $coli$ O157:H7 by 27.1, 67.7, and 83.3%, respectively. According to the results of the present study, administration of CGSCS10 to mice can reduce the severity of $E.$ $coli$ O157:H7 infection. In addition, it is suggested that CGSCS10 represents a good candidate for the treatment of enteric infections in domestic animals.

Susceptibility tests of oral antibiotics including cefixime against Escherichia coli, isolated from pediatric patients with community acquired urinary tract infections (소아 원외 요로감염 환아에서 분리된 E. coli에 대한 cefixime을 포함한 경구 항생제의 감수성 연구)

  • Lee, Soo Young;Lee, Jung Hyun;Kim, Jong Hyun;Hur, Jae Kyun;Kim, Sun Mi;Ma, Sang Hyuk;Kang, Jin Han
    • Clinical and Experimental Pediatrics
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    • v.49 no.7
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    • pp.777-783
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    • 2006
  • Purpose : Urinary tract infection(UTI) is one of the most frequent infections in children. E. coli is the most frequent etiological micropathogen in pediatric community UTI, and E. coli has developed resistance to many antibiotics, highlighting the need for regular surveys of this organism resistant patterns in the community. The aim of this study was to determine the oral antibiotic susceptibility patterns of E. coli, isolated from pediatric patients with uncomplicated community acquired UTI. Methods : E. coli isolates, obtained from pediatric patients with uncomplicated community acquired UTI between October in 2004 to September in 2005. And minimal inhibitory concentrations(MICs) of oral aminopenicillins and beta-lactamase inhibnitors(ampicillin, amoxacillin, ampicillin-sulbactam), oral cephalosporins(cefaclor, cefixime) and sulfa drug(trimethoprime-sulfamethoxazole) were performed according to the National Committee for Clinical Laboratory Standards(NCCLS) guide line. Results : Total 211 organisms were isolated from pediatric out-patients with community UTI. E. coli was the most common organism(89 percent), followed by E. fecalis, Proteus species, S. aureus, M. morganii, and P. aeruginosa. The resistant rates of aminopenicillins and beta-lactamase inhibitors, cefaclor and sulfa drug to E. coli were very high. But, the resistant rate of cefixime was markedly low, and ESBL strains were isolated with small rates. Conclusion : Our study results suggest that aminopenicillins, cefaclor and sulfa drug may not be useful as first line empirical antibiotics to treat pediatric patients with community UTI in Korea. But, 3rd generation cephalosporin such as cefixime can be used as effective second line antibiotics after primary treatment failure, also may be useful as an empirical first line antibiotic. Finally, we conclude that a continuous surveillance study to monitor susceptibility patterns of E. coli in community UTI will be needed for the standard guide lines of empirical oral antibiotic treatment.

Prevalence and Classification of Escherichia coli Isolated from bibimbap in Korea (비빔밥에서 분리한 대장균의 오염도 조사 및 특성 연구)

  • Lee, Da-Yeon;Lee, Joo-Young;Wang, Hae-Jin;Shin, Dong-Bin;Cho, Yong-Sun
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.126-131
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    • 2015
  • Pathogenic Escherichia coli is recognized as an important cause of diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome worldwide. This study was conducted to investigate the prevalence E. coli contamination in the Korean traditional food bibimbap. E. coli were isolated from 84 of 1142 (7.3%) bibimbap investigated from 2005 to 2011. Antibiotic resistance profiling demonstrated that 6 of the 84 isolates (7.2%) showed multiple drug resistance. Fifteen virulence genes specific for pathogenic E. coli such as Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli (EAEC) were examined by multiplex PCR for mixed bacterial cultures derived from bibimbap samples. The EPEC virulence gene (ent) was detected in 5 strains (5.9%), while ETEC, EAEC, and EIEC were not detected. STEC serotypes O103 (1.2%), O91 (1.2%), and O128 (6.0%) were found, but other serogroups such as O26, O157, O145, O111 and O121 were not detecded. Automated Repetitive-Sequence-Based PCR analysis showed different patterns.

Antimicrobial Activities of Green Tea Extract and Fractions on the E. coli O157:H7 (Escherichia coli O157:H7균에 대한 녹차 추출물 및 분획물의 항균효과)

  • Cho Sun-Young;Choi Jae-Ho;Ham Seung-Shi;Oh Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.20 no.1
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    • pp.48-52
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    • 2005
  • This study was investigated to determine the antimicrobial activity of ethanol extract of green tea, and of solvent fractionated ethanol extract on Escherichia coli O157:H7. MIC value of ethanol extracts and solvent fractionated ethanol extract (hexane, ethyl-acetate, chloroform and water) were $2000{\mu}g/ml,\;0,\;500{\mu}g/ml,\;2000{\mu}g/ml,\;1000{\mu}g/ml$, respectively. The antimicrobial activity of ethyl acetate fraction against E. coli O157 :H7 ranged from $250-2000{\mu}g/disk$. The antimicrobial activity was significantly increased as concentration dependent. When E. coli O157:H7 was pH-adjusted TSB containing $1000{\mu}g/ml$ ethyl acetate, E. coli O157:H7 was significantly inhibited at initial pH of 10, whereas the pathogen grew well in the presence of pH 4.5-pH 9. The inhibitory effect of the ethyl acetate fraction on the growth of E. coli O157 :H7 was investigated. Growth of both strain in the tested conditions were rapidly occurred up to 12 h, but no growth was occurred in the presence of $250-1000{\mu}g/mL$ of ethyl acetate fraction for 72 h.

Functional Characterization of sll1556 of Synechocystis sp. PCC6803 as Type II Isopentenyl Diphosphate Isomerase (Type II Isopentenyl Diphosphate Isomerase로서 Synechocystis sp. PCC6803의 sll1556의 작용 특성)

  • Cho, Kab-Yeon
    • The Korean Journal of Food And Nutrition
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    • v.23 no.4
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    • pp.526-530
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    • 2010
  • Isopentenyl diphosphate(IPP) isomerization to dimethylallyl diphosphate(DMAPP) is an important step for the efficient production of isoprenoids such as lycopene, ${\beta}$-carotene, astaxanthin, etc. The type II isopentenyl diphosphate isomerase gene from Synechocystis sp. PCC6803(sll1556, Syidi2) was cloned and expressed in Escherichia coli $DH5{\alpha}$. When E. coli $DH5{\alpha}$ harboring lycopene synthesis genes, crtE, crtB, and crtI and mevalonate pathway genes, MvK1, MvK2, and Mvd, was cultured on LB medium containing mevalonate, the strain grew very slowly be due to the toxicity of isopentenyl diphosphate derived from mevalonate. When Syidi2 was introduced to E. coli $DH5{\alpha}$ harboring the lycopene synthesis genes and mevalonate pathway genes, growth on mevalonate medium was fully restored and the colony showed red color indicating lycopene formation. The growth rate of the mutant strain, E. coli $DH5{\alpha}$(idi::${\Delta}km$), was very slow because of IPP accumulation and DMAPP deprivation. Ultimately the idi mutant was complemented by introducing the Syidi2 gene.