• Title/Summary/Keyword: E-coli

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Development of Recombinant Escherichia coli Expressing Rhodotorula glutinis Epoxide Hydrolase (Rhodotorula glutinis의 epoxide hydrolase 고효율 발현 유전자 재조합 Escherichia coli 생촉매 개발)

  • Lee Soo-Jung;Kim Hee-Sook
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.415-419
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    • 2006
  • The epoxide hydrolase (EH) of Rhodotorula glutinis which has a high enantioselectivity against aromatic epoxide substrates was expressed to high levels in Escherichia coli based on codon usage. We analysed the Preference of codon usage between the yeast, R. glutinis, and bacteria, E. coli. E. coli, Rosetta(DE3)pLysS, harbors pRARE plasmid with tRNA genes for rare-codons was employed as a host strain. The recombinant E. coli expressing R. glutinis EH showed an enhanced enantioselective hydrolysis activity toward racemic styrene oxide. Enantiopure (S)-styrene oxide with a high enantiopurity of 99% ee (enantiomeric excess) was obtained from racemic substrates.

Enterotoxin Production and Plasmid Detection of Citrate Utilizing Escherichia coli Isolated from Cattle (우(牛) 유래(由來) Citrate이용(利用) 대장균(大腸菌)의 장독소(腸毒素) 산생능(産生能) 및 Plasmid DNA)

  • Chae, Tae-chul;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.28 no.1
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    • pp.59-65
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    • 1988
  • This paper deals with the 0 groups of citrate utilizing variants of Escherichia coli ($Cit^+$ E. coli) isolated from cattle, the production of colicin, hemolysin, K99 antigen, heat stable enterotoxin, and the isolation of plasmid DNA. Among 42 $Cit^+$ E. coli, 12 strains were 020, 9 strains 08, 5 strains 045, 3 strains 0115, 1 strain 064, 1 strain 0139 and remaining strains(11) were untypable. Thirty-nine(81.3%) out of 48 $Cit^+$ E. coli were produced colicin and 13(27.0%) were produced hemolysin. Of 12 $Cit^+$ E. coli bearing K99 antigen, 6(50.0%) were produced heat stable enterotoxin. In gel electrophoresis for the isolation of plasmid DNA, the number of plasmids varied from 1 to 7 in 10 $Cit^+$ E. coli. It's molecular weight ranged from 2 to 50 Mdalton, and 50 Mdalton plasmid was commonly existed in all strains.

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Antibiotic Resistance of Pathogenic Escherichia coli Isolated from Piglets with Diarrhea (설사자돈 유래 대장균의 항생물질 내성에 관하여)

  • 조광현;박노찬;권헌일;김이준;박덕상
    • Korean Journal of Veterinary Service
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    • v.15 no.2
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    • pp.134-143
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    • 1992
  • The present study was conducted to investigate the biochemical characteristics and anti-biotic resistance of Escherichia coli(E. coli) isolated from piglets with diarrhea in Kyongbuk province during the Period from February to November 1991. 368 E. coli strains were isolated from 382 piglets with diarrhea and the biochemical and cultural reaction were compared with the classification criteria of Edwards and Ewing. Tetracycline and sulfadimethoxine were found to be highly ineffective at in vitro inhibition of the E. coli of piglets origin. The majority of E. coli were susceptible to amikacin, chloramphenicol and gentamicine. 89 (89.0%) of 100 strains of E. coil were resistant to one or more drugs. The organisms resistant to 20 or 3 drugs were 54(60.6%) of 89 strains, whereas 16(17.9%) strains were found to be resistant to one drug. 55(61.8%) out of 89 drug resistance strains carried R factors($R^+$) which were transfer-able to the recipients by conjugation.

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Fed-batch Culture of Recombinant E.coli for the Production of Penicillin G Amidase (Penicillin G Amidase생산을 위한 재조합 대장균의 유가배양에 관한 연구)

  • Lee, Sang-Mahn
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.314-319
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    • 2008
  • Penicillin G amidase (PGA, benzylpenicillinaminohydrolase, EC 3.5.1.11) is industrially important enzyme which converts penicillin G to 6-aminopenicillanic acid (6-APA) and phenylacetic acid (PAA). The PGA in E. coli ATCC 11105 is secreted into the periplasm after removing signal sequences and becomes heterodimer which composed of two subunits, small subunit (24 kDa) and large subunit (65 kDa). In this study, the PGA gene was obtained from E. coli ATCC 11105 using PCR (polymerase chain reaction) technique. The active PGA was successfully secreated into periplasm in E. coli BL2 1(DE3) harboring pET-pga plasmid. The optimized fed-batch fermentation, consisting of a three-step shift of culture temperature from $37^{\circ}C$ to $22^{\circ}C$, gave a productivity of 19.6 U/mL with a cell growth of 62 O.D. at 600 nm.

E. coli Disinfection Using a Multi Plasma Reactor (멀티 플라즈마 반응기를 이용한 E. coli 소독)

  • Kim, Dong-Seog;Park, Young-Seek
    • Journal of Environmental Health Sciences
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    • v.39 no.2
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    • pp.187-195
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    • 2013
  • Objectives: For the practical application of the dielectric barrier discharge plasma reactor, a plasma reactor able to manage large volumes of water is needed. This study investigated the possibility of the practical application of a multi-plasma reactor which is a scaled-up version of a single plasma reactor. Methods: The multi-plasma reactor consists of several high-voltage transformers and plasma modules (discharge, ground electrodes and quartz dielectric tubes). The effects of water characteristics such as voltage (30-120 V), air flow rate (1-5 l/min), number of high-voltage transformers and plasma modules, and water quality on Escherichia coli (E. coli) disinfection and decrease of COD and $UV_{254}$ absorbance were investigated. Results: The experimental results showed that at a voltage of over 80 V, most of the E. coli were disinfected within 90 seconds. E. coli inactivation was not affected by the air flow rate. E. coli disinfection in the multiplasma process showed the traditional log-linear form of the disinfection curve. E. coli inactivation performance by transformer 3-Reactor 5 and transformer 3-Reactor 3 were similar. The disinfection performance of the UV process was affected by artificial sewage water. However, the plasma process was less affected by the artificial sewage within the standards for effluent water quality. Conclusions: Disinfection performance with several low voltages and plasma modules of three to five in number applied to the plasma process was higher than that concentrating a small amount of high voltage through a single plasma reactor. Removal of COD, $UV_{254}$ absorbance, and E. coli disinfection with the plasma process were better than with the UV process.

An Epidemiologic Study on Hosts and Pathogens of Urinary Tract Infection in Urban Children of Korea (2012-2017)

  • Yoo, Yeong Myong;Park, Byeong Sub;Lee, Shin Young;Park, Kyu Jung;Jung, Hyun Joo;Pai, Ki Soo
    • Childhood Kidney Diseases
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    • v.23 no.1
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    • pp.29-35
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    • 2019
  • Purpose: We aimed to determine characteristics of host, causative organisms, and antibiotic susceptibility of bacteria in pediatric patients with UTI living in metropolitan area of Korea. Methods: Retrospective investigation was done for the causative organisms of UTI in 683 pediatric cases treated at Ajou University Hospital from 2012 to 2017. Patients were classified into Escherichia coli and non-E. coli group, where E. coli group was subdivided into ESBL(+) and ESBL(-) groups based on whether the bacteria could produce extended spectrum beta-lactamase (ESBL). Antibiotic susceptibility of the causative organism was also determined. Results: A total of 683 UTIs occurred in 550 patients, of which 463 (67.8%) were first-time infection and 87 (32.2%) were recurrent ones (2-7 recurrences, 2.52 average), and 64.9% were male and 35.1% were female. The most common causative organism was E. coli (77.2%) and ESBL(+) E. coli was found in 126 cases. The susceptibility of E. coli to 3rd or 4th generation cephalosporin was relatively higher than that to ampicillin or amoxicillin/clavulanic acid. ESBL(+) E. coli showed higher resistance rate to 3rd or 4th generation cephalosporin than ESBL(-) E. coli. Conclusion: New treatment guideline should be considered due to the incidence of ESBL(+) E. coli increased up to one quarter of UTI cases.

Optimization of Electro-UV-Ultrasonic Complex Process for E. coli Disinfection using Box-Behnken Experiment (Box-Behnken법을 이용한 E. coli 소독에서 전기-UV-초음파 복합 공정의 최적화)

  • Kim, Dong-Seog;Park, Young-Seek
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.3
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    • pp.149-156
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    • 2011
  • This experimental design and response surface methodology (RSM) have been applied to the investigation of the electro-UV-ultrasonic complex process for the disinfection of E. coli in the water. The disinfection reactions of electro-UV-ultrasonic process were mathematically described as a function of parameters power of electrolysis ($X_1$), UV ($X_2$), and ultrasonic process ($X_3$) being modeled by use of the Box-Behnken technique, which was used for fitting 2nd order response surface model. The application of RSM yielded the following regression equation, which is empirical relationship between the residual E. coli number (Ln CFU) in water and test variables in coded unit: residual E. coli number (Ln CFU) = 23.69 - 3.75 Electrolysis - 0.67 UV - 0.26 Ultrasonic - 0.16 Electrolysis UV + 0.05 Electrolysis Ultrasonic + 0.27 $Electrolysis^2$ + 0.14 $UV^2$ - 0.01 $Ultrasonic^2$). The model predictions agreed well with the experimentally observed result ($R^2$ = 0.983). Graphical 2D contour and 3D response surface plots were used to locate the optimum range. The estimated ridge of maximum response and optimal conditions for residual E. coli number (Ln CFU) using 'numerical optimization' of Design-Expert software were 1.47 Ln CFU/L and 6.94 W of electrolysis, 6.72 W of UV and 14.23 W of ultrasonic process. This study clearly showed that response surface methodology was one of the suitable methods to optimize the operating conditions and minimize the residual E. coli number of the complex disinfection.

Prevalence of CTX-M-type Extended-Spectrum $\beta$-Lactamases Producing Escherichia coli and Klebsieilla pneumoniae Isolates in General Hospitals in 2005 (임상에서 분리된 CTX-M형 Extended-Spectrum $\beta$-Lactamases를 생산하는 Escherichia coli와 Klebsiella pneumoniae의 유행)

  • Kim, Yun-Tae;Kim, Tae-Un
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.342-351
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    • 2006
  • The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.

Multiplication of Escherichia coli DH5α::gfp on Strawberry Fruit Surface (딸기과실 표면에서 Eschercia coli DH5α::gfp 증식)

  • Yun, Hyejeong;Park, Kyeonghun;Ryu, Kyoung Yul;Yun, Jong-Chul;Kim, Byung Seok
    • Food Science and Preservation
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    • v.20 no.2
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    • pp.250-256
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    • 2013
  • To verify the multiplication of microorganisms on the surface of strawberries, the fate of E. coli $DH5{\alpha}::gfp$ at different temperatures, times and strawberry extract concentrations were measured. The population of E. coli $DH5{\alpha}::gfp$ rapidly increased by 7.36~7.78 log CFU/g at $25{\sim}30^{\circ}C$ for 24 hr and slowly increased by 6.49~8.49 log CFU/g at $10{\sim}20^{\circ}C$ for 48 hr. However, E. coli $DH5{\alpha}::gfp$ did not grow at $10{\sim}15^{\circ}C$ on the surface of the strawberries, regardless of the contact times with the bacterial suspension. E. coli $DH5{\alpha}::gfp$ reached 1.52~3.26 log CFU/g at $20^{\circ}C$ as the contact frequency increased from two to six times. The contact frequencies did not significantly differ. In the case of the six-time contact on the surface of the strawberry at 25 and $30^{\circ}C$, the E. coli $DH5{\alpha}::gfp$ increased by 5.17 and 5.01 log CFU/g. The effects of the strawberry extracts on the growth of E. coli $DH5{\alpha}::gfp$ showed that sterilization and non-sterilization do not affect the growth of microorganisms for 96 hr. In the minimal broth, the growth of E. coli $DH5{\alpha}::gfp$ increased by 1 log CFU/g for 96 hr. In less than 50 percent of the strawberry extracts, the growth rate of E. coli $DH5{\alpha}::gfp$ was higher than in the control and increased by 4 and 5 log CFU/g at 50 and 25 percent of strawberry extracts, respectively. Therefore, E. coli $DH5{\alpha}::gfp$ can multiply and survive on the surface of strawberries when it comes into contact with the fruit extract.

Cloning of Bacillus amyloliquefaciens amylase gene using YRp7 as a vector I. Expression of cloned amylase gene in Escherichia coli (YRp 7 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I. Escherichia coli에서의 발현)

  • 서정훈;김영호;전도연;홍순덕;조윤래
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.161-168
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    • 1986
  • A 1.95Kb Sau3Al fragment coding for $\alpha$-amylase from Bacillus amyloliquefaciens was isolated by the shotgun method using Escherichia coli as a host. The genome of Bacillus amyloliquefaciens was partially digested with the restriction endonuclease Sau3Al and joined to plasmid YRp7 cleaved with the restriction endonuclease BamHI. The $\alpha$-amylase gene present in a 1.95Kb insert was stably maintained and expressed in Escherichia coli. The amount of $\alpha$-amylase activity produced by Escherichia coli containing the hybrid plasmid pEA24 was about 65% of the activity produced by the donor Bacillus amyloliquefaciens strain. The properties of $\alpha$-amylase produced by Escherichia coli were very similar to those produced by Bacillus amyloliquefaciens as based on optimum temperature, pH, and effect of CaCl$_2$ concentration. About 70% of the $\alpha$-amylase produced by Escherichia coli was localized in the periplasmic space, whereas the remaining enzyme was localized in the inner part of the cell.

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