• Title/Summary/Keyword: C. coli

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5-Aminolevulinic Acid Biosynthesis in Escherichia coli Coexpressing NADP-dependent Malic Enzyme and 5-Aminolevulinate Synthase

  • Shin, Jeong-Ah;Kwon, Yeong-Deok;Kwon, Oh-Hee;Lee, Heung-Shick;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1579-1584
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    • 2007
  • 5-Aminolevulinate (ALA) synthase (E.C. 2.3.1.37), which mediates the pyridoxal phosphate-dependent condensation of glycine and succinyl-CoA, encoded by the Rhodobacter sphaeroides hemA gene, enables Escherichia coli strains to produce ALA at a low level. To study the effect of the enhanced C4 metabolism of E. coli on ALA biosynthesis, NADP-dependent malic enzyme (maeB, E.C. 1.1.1.40) was coexpressed with ALA synthase in E. coli. The concentration of ALA was two times greater in cells coexpressing maeB and hemA than in cells expressing hemA alone under anaerobic conditions with medium containing glucose and glycine. Enhanced ALA synthase activity via coupled expression of hemA and maeB may lead to metabolic engineering of E. coli capable of large-scale ALA production.

Study of Anti-bacterial Properties for Impregnated Activated Carbon by Silver Nano-particles (은나노 입자가 첨착된 활성탄의 항균특성에 관한 연구)

  • Lee, Chul-Jae;Kim, Dong-Yeub;Kim, Byung-So
    • Applied Chemistry for Engineering
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    • v.18 no.4
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    • pp.396-399
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    • 2007
  • In present work, the anti-bacterial effect of silver/activated carbon (Ag/C) composites prepared by the ${\gamma}$-irradiation of $AgNO_3$ solution on Escherichia coli (E. coli) has been studied. Characteristics of the Ag/C composites were identified by scanning electron microscopy (SEM), X-ray diffraction (XRD) and atomic absorption spectroscopy (AAS). The inhibitory concentration of E. coli was found to be 0.387 ppm and the sterilizing concentration for the tested organism was 1.017 ppm. These results support the evidence that Ag/C composites have strong antibacterial activity to E. coli.

Antimicrobial Properties of Turmeric (Curcuma longa L.) Rhizome-Derived ar-Turmerone and Curcumin

  • Lee, Hoi-Seon
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.559-563
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    • 2006
  • The growth responses of six bacterial strains exposed to materials extracted from turmeric (Curcuma longa) rhizomes were examined using impregnated paper disk agar diffusion. Methanol extracts of turmeric rhizomes exhibited strong inhibitory activity against Clostridium perfringens and weak inhibitory activity toward Escherichia coli at 5 mg/disk. However, in tests conducted with Bifidobacterium adolescentis, B. bifidum, B. longum, and Lactobacillus casei, the methanol extract showed no inhibitory response. The biologically active constituent isolated from the turmeric rhizomes extracts was characterized as ar-turmerone using various spectroscopic analyses including EI-MS and NMR. The responses varied according to the dosage, chemicals, and bacterial strain tested. At 2 and 1 mg/disk, ar-turmerone strongly inhibited the growth of C. perfringens and moderately inhibited the growth of E. coli without any adverse effects on the growth of four lactic acid-bacteria. Of the commercially available compounds originating from turmeric rhizomes, curcumin exhibited strong and moderate growth inhibition against C. perfringens at 2 and 1 mg/disk, respectively, and weak growth inhibition against E. coli at 1 mg/disk. However, little or no activity was observed for borneol, 1,8-cineole, and sabinene against all six bacteria strains tested. The observed inhibitory activity of the turmeric rhizome-derived curcumin and ar-turmerone against C. perfringens and E. coli demonstrate one of the important pharmacological activities of turmeric rhizomes.

Development and Characterization of Expression Vectors for Corynebacterium glutamicum

  • Lee, Jinho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.70-79
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    • 2014
  • In an attempt to develop a variety of expression vector systems for Corynebacterium glutamicum, six types of promoters, including $P_{tac}$, $P_{sod}$, $P_{sod}$ with a conserved Shine-Dalgarno (SD) sequence from C. glutamicum, $P_{ilvC}$, $P_{ilvC}$ with a conserved SD-1 ($P_{ilvC-M1}$), and $P_{ilvC}$ with a conserved SD-2 ($P_{ilvC-M2}$), were cloned into a modified shuttle vector, pCXM48. According to analysis of promoter strength by quantitative reverse transcription PCR, $P_{sod}$ and $P_{sod-M}$ were superior to tac and ilvC promoters in terms of transcription activity in C. glutamicum. All of the promoters have promoter activities in Escherichia coli, and $P_{sod-M}$ displayed the highest level of transcriptional activity. The protein expression in constructed vectors was evaluated by measuring the fluorescence of green fluorescent protein (GFP) and SDS-PAGE. C. glutamicum harboring plasmids showed GFP fluorescence with an order of activity of $P_{ilvC}$ > $P_{ilvC-M1}$ > $P_{sod}$ > $P_{ilvC-M2}$ > $P_{sod-M}$, whereas all plasmids except pCSP30 with $P_{sod}$ displayed fluorescence activities in E. coli. Of them, the strongest level of GFP was observed in E. coli with $P_{sod-M}$, and this seems to be due to the introduction of the conserved SD sequence in the translational initiation region. These results demonstrate that the expression vectors work well in both C. glutamicum and E. coli for the expression of target proteins. In addition, the vector systems harboring various promoters with different strengths, conserved SD sequences, and multiple cloning sites will provide a comfortable method for cloning and gene expression, and consequently contribute to the metabolic engineering of C. glutamicum.

Survival of Escherichia coli O157:H7 and Salmonella ser. typhimurium in Fermented Milk Products (발효유제품내에서 Escherichia coli O157:H7과 Salmonella ser. typhimurium의 생존)

  • 김현욱;안영태;임정현;강호진;장영호
    • Journal of Food Hygiene and Safety
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    • v.12 no.3
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    • pp.175-180
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    • 1997
  • Escherichia coli O157:H7 and Salmonella ser. typhimurium are pathogens involved in food poisoning in numerous countries. This study aimed to obtain knowleges on the survival of E. coli O157:H7 KSC 109 and S. ser. typhimurium ATCC 14028 in fermentedmilk products which were on sale in Suwon Yakult supplier. To the final concentration of 103~104 cfu/$m\ell$ of E. coli O157:H7 KSC 109 or S. wer. typhimurium ATCC 14028 in the fermented milks, Metchnikoff, Ace, Yakult, Mastoni and Super 100 were inoculated with these pathogens and then were stored at 4$^{\circ}C$ and viable cells of these pathogens were periodically counted. The results showed that the survival of two pathogens differed in the different types of fermented milks tested. Number of suriviving E. coli O157:H7 KSC 109 and S. ser. typimurium ATCC 14028 cells (initial inoculum, 103~104 cfu/$m\ell$) were decreased to 101, 102 cfu/$m\ell$ in Ace after 100 hours, and were decreased gradually to 101 cfu/$m\ell$ in Yakult after 250 hours. In the other fermented milks, viable cells of E. coli O157:H7 KSC 109 was not drastically decreased but those of S. ser. typhimurium ATCC 14028 was decreased gradually to 102 (Mastoni), and to 101 cfu/$m\ell$ (Super 100) after 250 hours. It appeared that S. ser. typhimurium ATCC 14028 was more susceptible than E. coli O157:H7 KSC 109 at low pH. Vibale cells of E. coli O157:H7 KSC 109 was not drastically decreased in most of fermented milks tested except Ace and Yakult, but in general, S. ser. typhimurium ATCC 14028 was drastically decreased in most of the fermented milks. The major ingibition factor against these pathogens in the fermented milks during storage at 4$^{\circ}C$ appeared to be the acidity and the metabolites produced by the starters bacteria used in fermented milk products.

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Prevalence of virulence and cytolethal distending toxin (CDT) genes in thermophilic Campylobacter spp. from dogs and humans in Gyeongnam and Busan, Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.54 no.1
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    • pp.39-48
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    • 2014
  • The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barr$\acute{e}$ syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted $R^2$= 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

Repression of Escherichia coli serC-aroA Operon by Aromatic Amino Acids (방향족 아미노산에 의한 대장균 serC-aroA Operon의 발현 억제)

  • Hwang, Woo-Gil;Sa, Jae-Hoon;Kim, Kyung-Hoon;Lim, Chang-Jin
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.109-114
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    • 1994
  • The Escherichia coli aroA and serC genes constitute a mixed-function operon which involves in two different amino acid biosynthetic pathways. The regulation of expression of serC-aroA operon was evaluated through the use of a serC-araA-lacZ fusion plasmid pWH2. The expression of the serC-aroA operon was decreased by aromatic amino acids such as tyrosine, tryptophan, and phenylalanine. The repressible effects were diminished in E. coli tyrR of trpR strain, indicating the involvemnt of TyrR of TrpR protein in the repression. Tyrosine was competitie with cAMP in the influence on the expression of the serC-AroA operon. From these data, it was suggested that the serC-aroA operon is controlled by aromatic amino acids in a negative manner.

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Inverse Correlation between Extracellular DNase Activity and Biofilm Formation among Chicken-Derived Campylobacter Strains

  • Jung, Gi Hoon;Lim, Eun Seob;Woo, Min-Ah;Lee, Joo Young;Kim, Joo-Sung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.1942-1951
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    • 2017
  • Campylobacter jejuni and Campylobacter coli are important foodborne pathogenic bacteria, particularly in poultry meat. In this study, the presence of extracellular DNase activity was investigated for biofilm-deficient Campylobacter strains versus biofilm-forming Campylobacter strains isolated from chickens, to understand the relationship between extracellular DNase activity and biofilm formation. A biofilm-forming reference strain, C. jejuni NCTC11168, was co-incubated with biofilm non-forming strains isolated from raw chickens or their supernatants. The biofilm non-forming strains or supernatants significantly prohibited the biofilm formation of C. jejuni NCTC11168. In addition, the strains degraded pre-formed biofilms of C. jejuni NCTC11168. Degradation of C. jejuni NCTC11168 biofilm was confirmed after treatment with the supernatant of the biofilm non-forming strain 2-1 by confocal laser scanning microscopy. Quantitative analysis of the biofilm matrix revealed reduction of extracellular DNA (16%) and proteins (8.7%) after treatment. Whereas the biofilm-forming strains C. jejuni Y23-5 and C. coli 34-3 isolated from raw chickens and the C. jejuni NCTC11168 reference strain showed no extracellular DNase activity against their own genomic DNA, most biofilm non-forming strains tested, including C. jejuni 2-1, C. coli 34-1, and C. jejuni 63-1, exhibited obvious extracellular DNase activities against their own or 11168 genomic DNA, except for one biofilm non-former, C. jejuni 22-1. Our results suggest that extracellular DNase activity is a common feature suppressing biofilm formation among biofilm non-forming C. jejuni or C. coli strains of chicken origin.

Growth Inhibition of Listeria monocytogenes and Escherichia coli O157:H7 in Naengmyon-Broth by addition of Antibacterial Dongchimi-Juice (항균성 동치미액의 첨가에 의한 냉면국물 중의 Listeria monocytogenes 및 Escherichia coli O157:H7 생육억제)

  • 박상희
    • The Korean Journal of Food And Nutrition
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    • v.12 no.2
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    • pp.133-141
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    • 1999
  • Juice of Dongchime a Korean traditional vegetable food fermented with lactic acid bacteria has been traditionary used as broth for Naengmyon a Korean cold noodles with broth. This study was carried out to demonstrate the growth inhibition of two food born pathogens Listeria monocytogenes and Escherichia coli O157:H7 in Naengmyon-broth containing Dongchimi-juice fermented with high antibacterial lactic acid bacteria Lactobacillus homohiochii B21 and Leuconostoc mesenteroides C16. Naengmyon-broth were made with beef broth and Dongchimi-juice fermented with lactic acid bacteria and the changes in viable cell counts of the inoculated pathogens in Naengmyon-broths were investigated during storage at 2$0^{\circ}C$ and 1$0^{\circ}C$. In Naengmyon-broth of 100% Dongchimi-juice stored at 2$0^{\circ}C$ the numbers of Listeria monocyto-genes and Escherichia coli O157:H7 were rapidly decreased from 106CFU/ml to 106CFU/ml in 8 hours and 40 hours respectively. In Naengmyon-broth containing 50% Dongchimi-juice their numbers were also rapidly decreased though the decreasing rates were not so fast as those in 1005 Dongchimi-juice. In Naengmyon-broth containing 10% Dongchimi-juice the growths of the two pathogens were markedly inhibited compared with those in 100% beef broth though some growths were occurred in early phase. But in Naengmyon-broth of 100% beef broth their growths were very fast from early. Antibacterial ac-tivity of the Dongchimi-juice was more distinct at 2$0^{\circ}C$ than at 1$0^{\circ}C$ and was more active against Listeria monocytogenes than against Escherichia coli 157:H7.

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Cloning, Nucleotide Sequence and Expression of Gene Coding for Poly-3-hydroxybutyric Acid (PHB) Synthase of Rhodobacter sphaeroides 2.4.1

  • Kim, Ji-Hoe;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.7 no.4
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    • pp.229-236
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    • 1997
  • A gene, $phbC_{2.4.1}$ encoding poly-3-hydroxybutyric acid (PHB) synthase of Rhodobacter sphaeroides 2.4.1 was cloned by employing heterologous expression in Escherichia coli. R. sphaeroides chromosomal DNA partially digested with MboI was cloned in pUC19 followed by mobilization into E. coli harbouring $phbA,B_{AC}$ in pRK415, which code for ${\beta}$-ketothiolase and acetoacetyl CoA reductase of Alcaligenes eutrophus, respectively. Two E. coli clones carrying R. sphaeroides chromosomal fragment of $phbC_{2.4.1}$ in pUC19 were selected from ca. 10,000 colonies. The PHB-producing colonies had an opaque white appearance due to the intracellular accumulation of PHB. The structure of PHB produced by the recombinant E. coli as well as from R. sphaeroides 2.4.1 was confirmed by [$H^{+}$]-nuclear magnetic resonance (NMR) spectroscopy. Restriction analysis of the two pUC19 clones revealed that one insert DNA fragment is contained as a part of the other cloned fragment. An open reading frame of 601 amino acids of $phbC_{2.4.1}$ with approximate M.W. of 66 kDa was found from nucleotide sequence determination of the 2.8-kb SaiI-PstI restriction endonuclease fragment which had been narrowed down to support PHB synthesis through heterologous expression in the E. coli harbouring $phbA,B_{AC}$. The promoter (s) of the $phbC_{2.4.1}$ were localized within a 340-bp DNA region upstream of the $phbC_{2.4.1}$ start codon according to heterologous expression analysis.

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