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Enhanced Heterologous Expression of Aspergillus niger Epoxide Hydrolase and Its Application to Enantioselective Hydrolysis of Racemic Epoxides (Aspergillus niger의 Epoxide Hydrolase 고효율 발현 및 라세믹 에폭사이드의 입체선택적 가수분해)

  • Lee, Soo Jung;Kim, Hee Sook;Lee, Eun Yeol
    • Applied Chemistry for Engineering
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    • v.17 no.5
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    • pp.557-560
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    • 2006
  • The epoxide hydrolase (EH) of Aspergillus niger LK was expressed to high levels in Escherichia coli based on codon usage. E. coli, Rosetta (DE3)PLysS, containing a large number of tRNAs for rare-codons was employed as a host strain. The recombinant E. coli expressing A. niger EH showed an enhanced enantioselective hydrolysis activity toward racemic styrene oxide. Enantiopure (S)-styrene oxide with a high enantiopurity of 99% ee was obtained from racemic substrates.

Antibacterial Activity of Powdered Spice against Escherichia coli and Staphylococcus aureus (향신료 분말의 Esdcherichia coli 와 Staphylococcus aureus 에 대한 항균작용)

  • 김미림;최경호;박찬성
    • Food Science and Preservation
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    • v.7 no.1
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    • pp.124-131
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    • 2000
  • Antibacterial activities of powdered spices(garlic , ginger, cinnamon and clove) against pathogenic Escherichia coli )157:H7 and Staphyloccus auresus were investigated. Spice powder was added in was exponetial phase of each bacterial culture . Growth inhibition was determined by the absorbance at 660nm and morphological changes of the cells were observed by transmission electron microscope (TEM). Ginger powder has the highest antibacterial activity, following cinnamon , clove and garlic has the least activity.Growth of Escherichia coli O157:H7 and Staphyloccus aureus were completely inhibited within 5 hours after addition of 1 % of garlic , 0.3% of ginger or cinnamon , 0.5% of clove powder on the exponential phase of the cells. Spice untreated cells of E. coli and S. aureus, the cytoplasm was entirely surrounded by rigid cell wall and cell walls formed a smooth layer well attached to the plasma membrane. In the cells of E. coli and S. aureus treated with spice powder, cell wall and plasma membrane were lysed and severely damaged. E.coli cells growth in the presence of spice powder showed plammolysis, the loss of electron dense material, the formation of extra cellular blebs and cytoplasm burst out from the cell. S .sureus cells grown in the presence of spice powder showed swell of cell wall, the loss of electron dense material , coagulation of cell cytoplasm and formation of extra cellular blebs. Severely damaged cells of S. aureus lost whole cytoplasm and left as ghost of the cell. Spice powder stimulated autolyssi and induced cell death.

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Stability of Recombinant Plasmids Carrying the stb Locus of E. coli IncFII NR1 Plasmid in E. coli and Yeast (대장균과 효모에서 Escherichia coli IncFII NR1 플라스미드의 stb 좌위를 포함하는 재조합 플라스미드의 안정성에 관한 연구)

  • Chung, Kung-Sook;Kim, Choon-Kwang;Kim, Kyu-Won
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.37-43
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    • 1993
  • The effect of stb locus of E. COLI IncFII plasmid NR1 on the stability of chimeric plasmids was investigated. First, we have isolated the stability locus (stb) from E. coli NR1 plasmid and then inserted into the three different vectors, pUC8, YRp17 and YEp24. By examining their stability in E. coli and yeast, we showed that the recombinant plasmids containing stb locus were resonably stable. Also, by comparing the amounts of the rDNA fragments per haploid genome with those of the plasmid fragments, we showed they copy number of recombinant plasmids was not increased. Consequently, the stb locus of E. coli IncFII plasmid NR1 stabilized the chimeric plasmids but did not affect the replication or copy number of plasmids.

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ATP-independent Thermoprotective Activity of Nicotiana tabacum Heat Shock Protein 70 in Escherichia coli

  • Cho, Eun-Kyung;Bae, Song-Ja
    • BMB Reports
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    • v.40 no.1
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    • pp.107-112
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    • 2007
  • To study the functioning of HSP70 in Escherichia coli, we selected NtHSP70-2 (AY372070) from among three genomic clones isolated in Nicotiana tabacum. Recombinant NtHSP70-2, containing a hexahistidine tag at the amino-terminus, was constructed, expressed in E. coli, and purified by $Ni^{2+}$ affinity chromatography and Q Sepharose Fast Flow anion exchange chromatography. The expressed fusion protein, $H_6NtHSP70$-2 (hexahistidine-tagged Nicotiana tabacum heat shock protein 70-2), maintained the stability of E. coli proteins up to 90$^{\circ}C$. Measuring the light scattering of luciferase (luc) revealed that NtHSP70-2 prevents the aggregation of luc without ATP during high-temperature stress. In a functional bioassay (1 h at 50$^{\circ}C$) for recombinant $H_6NtHSP70$-2, E. coli cells overexpressing $H_6NtHSP70$-2 survived about seven times longer than those lacking $H_6NtHSP70$-2. After 2 h at 50$^{\circ}C$, only the E. coli overexpressing $H_6NtHSP70$-2 survived under such conditions. Our NtHSP70-2 bioassays, as well as in vitro studies, strongly suggest that HSP70 confers thermo-tolerance to E. coli.

Electric Field Analysis and Removal Characteristics of Escherichia Coli for Water Discharge Tube with Globular SiO$_2$ (구형 SiO$_2$를 갖는 수방전관의 전계 해석 및 대장균 제거 특성)

  • 이동훈
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.53 no.2
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    • pp.103-108
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    • 2004
  • This paper shows the simulation of electric field distribution and removal characteristics of Escherichia coli for water discharge tube with globular $SiO_2$. At the experiments of the removing Escherichia coli used the discharge tube with globular dielectric($SiO_2$), because the electric field is increased when applied voltage is increased, the removed ratio of Escherichia coli was related with increasing of applied voltage. When a passing number of test water in water discharge tube is increased, the removed ratio of Escherichia coli is increased because passing number of territory with electric field is increased. When diameter of globular dielectric($SiO_2$) is increased, the removed time of Escherichia coli was decreased because electric field for dielectric polarization of globular dielectric($SiO_2$) was increased. Also, the removed ratio of Escherichia coli of the water discharge tube with globular dielectric($SiO_2$) was measured higher than the removal ratio of the discharge tube without globular dielectric($SiO_2$)

Characterization of Diarrheagenic Escherichia coli Isolated from Fresh Beef, Pork, and Chicken Meat in Korean Markets

  • Cho, Yong Sun;Koo, Min Seon;Jang, Hye Jin
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.121-128
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    • 2020
  • Diarrhea is a major public health concern associated with pathogenic Escherichia coli infections. Food-borne pathogenic E. coli can lead to large diarrheal outbreaks and hence, there is a need to estimate the frequency of pathogenic E. coli load in the various types of meat available in markets. In the present study, we classified and characterized diarrheagenic E. coli isolates collected from 399 raw meat samples from retail sources in Korea. Shiga toxin-producing E. coli (STEC) were detected in 11 (9.7%) samples, including nine strains (8.0%) in beef and two strains (1.8%) in chicken. The frequency of the detected virulence markers were as follows: astA, 28.3%; escV,18.6%; eaeA,17.7%; ent, 7.0%; EHEC-hly, 4.4%; stx1, 3.5%; and stx2, 3.5%. We did not observe any typical EPEC, EIEC, or ETEC virulence determinants in any of the samples. The STEC serotype O26 was detected in one sample, but no other serogroups (O91, O103, O128, O157, O145, O111, and O121) were found. Further research is needed to better understand the virulence mechanism of STEC serotypes, their ecology, and prevalence in animals, food, and the environment. These results will help improve risk assessment and predict the sources of food poisoning outbreaks.

Effect of Bifidobacterium longum on Growth Inhibition of Enterotoxigenic Escherichia coli $A_2$ (장내세균 Bifidobacterium longum에 의한 병원성 Escherichia coli $A_2$의 생육저해)

  • 성문희;신현정;강국희
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.203-207
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    • 1985
  • Bifidobacteria are normal inhabitants of the intestinal tract of humans. Using Bif. longum isolated from feces of Korean adult and Bifidus preparation, we observed the growth inhibitory actions of these organisms toward E. coli $A_2$ causing bacterial diarrhea. Bif. longum SKD-2001 SKD-2004 inhibited the growth of E. coli $A_2$ drastically. It is supposed that the mechanism of the growth inhibitory actions is due to acid conditions created by Bif. longum.

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Removal characteristic of Escherichia coli for Discharge tube with globular $SiO_2$ (구형 $SiO_2$를 갖는 방전관의 수중 세균제거특성)

  • Lee, Dong-Hoon;Park, Hong-Jae;Park, Jae-Youn
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2003.11a
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    • pp.546-549
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    • 2003
  • This paper was shown removal characteristic of escherichia coli for discharge tube with globular $SiO_2$. At the result of the removal characteristic experiments of escherichia coli using the discharge tube with globular $SiO_2$, because the electric field is also increased when input voltage is increased, the removal characteristic of escherichia coli was appeared relation connection to input voltage. If a passing number of test water in discharge tube is increased, the removal ratio of escherichia coli is increased because passing number of electric field section is increased. When diameter of globular $SiO_2$ is increased, the removal time of escherichia coli is to be decreased because dielectric polalization of globular dielectric($SiO_2$), Also, the removal ratio of escherichia coli of the discharge tube with globular dielectric($SiO_2$) is appeared higher than the removal ratio of the discharge tube without globular dielectric($SiO_2$).

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Inactivation of Escherichia coli and MS2 coliphage by Cu(II)-activated peroxomonosulfate in natural water

  • Kim, Hyung-Eun;Lee, Hye-Jin;Kim, Min Sik;Choi, Joon-Young;Lee, Changha
    • Membrane and Water Treatment
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    • v.10 no.3
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    • pp.231-237
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    • 2019
  • Peroxymonosulfate (PMS) in combination with Cu(II) was examined to inactivate E. coli and MS2 coliphage in natural water. The combined system (i.e., the Cu(II)/PMS system) caused a synergistic inactivation of E. coli and MS2, in contrast with either Cu(II) or PMS alone. Increasing the concentration of PMS enhanced the inactivation of E. coli and MS2, but after a certain point, it decreased the efficacy of the microbial inactivation. In the Cu(II)/PMS system, adding reactive oxidant scavengers marginally affected the E. coli inactivation, but the inhibitory effects of copper-chelating agents were significant. Fluorescent assays indicated that the Cu(II)/PMS system greatly increased the level of reactive oxidants inside the E. coli cells. The sequential addition of Cu(II) and PMS inactivated more E. coli than did adding the two simultaneously; in particular, the inactivation efficacy was much higher when Cu(II) was added first. The observations from the study collectively showed that the microbial inactivation by the Cu(II)/PMS system could be attributed to the toxicity of Cu(I) as well as the intracellular oxidative stress induced by Cu(III) or radical species.

Molecular characteristics of ESBL-producing Escherichia coli isolated from chickens with colibacillosis

  • Yoon, Sunghyun;Lee, Young Ju
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.37.1-37.8
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    • 2022
  • Background: Avian pathogenic Escherichia coli (APEC) causes colibacillosis, resulting in significant economic losses in the poultry industry. Objectives: In this study, the molecular characteristics of two extended-spectrum beta-lactamase (ESBL)-producing APEC isolates were compared with previously reported ESBL-producing E. coli isolates. Methods: The molecular characteristics of E. coli isolates and the genetic environments of the ESBL genes were investigated using whole genome sequencing. Results: The two ESBL-producing APEC were classified into the phylogenetic groups C and B1 and ST410 and ST162, respectively. Moreover, the ESBL genes of the two isolates were harbored in different Inc plasmids. The EC1809182 strain, harboring the blaCTX-M-55 gene on the plasmid, exhibited extensive homology to IncFIB (98.4%) and IncFIC(FII) (95.8%). The EC1809191 strain, harboring the blaCTX-M-1 gene, was homologous to IncI1-I (Gamma) (99.3%). All chromosomes carried the multidrug transporter, mdf(A) gene. Mobile genetic elements, adjacent to CTX-M genes, facilitated the dissemination of genes in the two isolates, analogous to other ESBL-producing E. coli isolates. Conclusions: This study clarifies the transmission dynamics of CTX-M genes and supports strengthened surveillance to prevent the transmission of the antimicrobial-resistant genes to humans via the food chain.