• Title/Summary/Keyword: inactivation of E. coli

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Purification and Characterization of Aminoglycoside-Resistant inhibitior from methylotrophic Actinomycetes (Methanol 자화 방선균으로부터 Aminoglycoside 내성 저해물질의 정제 및 특성)

  • 김현수;신재욱
    • Microbiology and Biotechnology Letters
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    • v.27 no.3
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    • pp.215-222
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    • 1999
  • Methylotrophic actinomycetes No. 155 produced an aminoglycoside antibiotics(AG)-resistant inhibitor. We have previously reported that the inhibitor shows strong inhibition to sisomicin-resistant strain. In order to understand the functions of inhibitor and sisomicin-resistance, characterizations and purification of inhibitor were investigated. Strain No. 155 was tentatively identified as Nocardiopsis sp. based on morphological and some physiological characteristics. In the antimicrobial activity test, the addition of inhibitor to sisomicin showed a reduction effect of MIC on the test strains such as Gram(+), Gram(-) bacteria and yeasts. The combination of the inhibitor and various antibiotics revealed synergistic against E. coli K-12 and B. subtilis PCI 219. The induced intracellular proteins from sisomicin-resistant strain exhibited the sisomicin inactivation by invitro test. And the induced intracellular proteins were inactivated by addition of the inhibitor. The inhibitor compound was purified by anion exchange chromatography(Dowex-1) and HPLC using Asahipak ES-502C column. The purified inhibitor compound was detected in a single peak(above 98.5% purity) through the HPLC analysis.

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Helium dielectric barrier discharge-cold plasma treatment for microbiological safety and preservation of onion powder (유전체 방벽 방전 콜드 플라즈마 기술을 이용한 양파 분말 미생물 안전성 향상 및 품질 보존)

  • Won, Mee Yeon;Choi, Ha Young;Lee, Kwang Sik;Min, Sea Cheol
    • Korean Journal of Food Science and Technology
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    • v.48 no.5
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    • pp.486-491
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    • 2016
  • Efficacy of dielectric barrier discharge-cold plasma treatment (DBD-CPT) for microbial decontamination of onion powder was evaluated. Onion powder, inoculated with Escherichia coli O157:H7, Salmonella Enteritidis, or Listeria monocytogenes, was treated with helium DBD-CPT. DBD-CPT (9 kV, 20 min) inhibited E. coli O157:H7, S. Enteritidis, and L. monocytogenes by $1.4{\pm}0.5$, $2.3{\pm}0.3$, and $1.2{\pm}0.0log\;CFU/cm^2$, respectively. The inactivation levels of E. coli O157:H7, S. Enteritidis and L. monocytogenes increased by $2.2{\pm}0.1$, $2.5{\pm}0.1$ and $1.9{\pm}0.3log\;CFU/cm^2$, respectively, as water activity increased from 0.4 to 0.8, and increased by $2.3{\pm}0.4$, $2.1{\pm}0.1$ and $1.6{\pm}0.1log\;CFU/cm^2$, respectively, as the particle size increased from 0.3 to $1.0cm^2$. Neither the ascorbic acid and quercetin concentrations nor the color of onion powder was changed by DBD-CPT (p>0.05). These results demonstrate the potential for application of DBD-CPT in improving microbiological safety of onion powder while preserving the physicochemical properties.

Expression and Characterization of Thiol-Specific Antioxidant Protein, DirA of Corynebacterium diphtheriae (코리네박테리움 디프테리아 티올 특이성 항산화단백 DirA의 발현 및 특성)

  • Myung-Jai Choi;Kanghwa Kim;Won-Ki Choi
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.1-9
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    • 1998
  • A Corynebacterium diphtheriae iron-repressible gene dirA, that was homologous to TSA of Saccharomyces cerevisiae and AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase, was amplified with PCR and expressed in E. coli. The DirA purified from the transformed E. coli crude extracts prevented the inactivation of enzyme caused by metal-catalyzed oxidation (MCO) system containing thiols but not by ascorbate/Fe$^{3+}$/$O_2$ MCO system. The DirA concentration, which inhibited the inactivation of glutamine synthetase by 50% (IC$_{50}$) against MCO system, was 0.12 mg/ml. The multimeric forms of DirA were converted to the monomeric form in SDS-PAGE under the thioredoxin system comprised of NADPH, Saccharomyces cerevisiae thioredoxin reductase, and thioredoxin. Also, DirA showed thioredoxin dependent peroxidase activity. All of these results were consistent with the characteristics of a thiol specific antioxidant (TSA) protein having two conserved cysteine residues.

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Sterilization of Rapeseed Sprouts by Intense Pulsed Light Treatment (고강도 광원을 이용한 새싹 채소의 살균)

  • Park, Heeran;Cha, Gyung-Hee;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.48 no.1
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    • pp.36-41
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    • 2016
  • In this study, the effects of intense pulsed light (IPL) treatment on microbial inactivation and quality in rapeseed sprouts were investigated. Untreated rapeseed sprouts exhibit a high level of total aerobic bacteria (TAB) ($1.2{\times}10^7CFU/g$), coliform bacteria (coliform) ($3.3{\times}10^6CFU/g$), and pathogenic E. coli (PE) ($2.1{\times}10^5CFU/g$). The microorganisms found on rapeseed sprouts decreased with exposure to increasing light intensity and treatment time. The greatest reduction in microbial content was observed with a treatment of 1000 V, 5 pps for 10 min, where TAB, coliform, and PE levels decreased to 1.0 log CFU/g, 1.6 log CFU/g, and 1.8 log CFU/g, respectively. In agreement with these data, the microbial inactivation rate increased with the increase in the distance between the light source and the samples during IPL treatment. After IPL treatment of rapeseed sprouts, water content and vitamin C content decreased.

Removal Characteristics of Total Coliforms in a Rotating Activated Bacillus Contactor Process (회전식 부착 바실러스를 이용한 하수고도처리 공정에서의 총대장균군 제거 특성)

  • Kim, Eung-Ho;Cho, Yeon-Je;Park, Seong-Joo;Shin, Kwang-Soo;Yim, Soo-Bin;Park, Hyun-Ju
    • Journal of Korean Society on Water Environment
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    • v.21 no.1
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    • pp.73-78
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    • 2005
  • This study was performed to examine the disinfection capability of a Rotating Activated Bacillus Contactor (RABC) system, in which the predominant species, Bacillus sp. was expected to have a removal or inactivation effect of total coliforms. In a settling test with mixtures of E. coli and Bacillus sp., a high removal of E. coli was observed at $20{\sim}40^{\circ}C$, while insignificant removal at $10^{\circ}C$. In a batch test, a 4.5% addition of Bacillus sp. to activated sludge considerably enhanced the removal effect of total coliforms, indicating Bacillus sp. played an important role in improving the settlability of the sludge and coliforms. In a pilot scale RABC system, the concentration of total colifroms reduced remarkably in the settling tank, suggesting that total coliforms in the RABC process were eliminated through coagulation and precipitation, probably due to extracellular polymeric substance (EPS) of Bacillus sp. The fraction of Bacillus sp. in the total cell count in the RABC process was in the range of 4.5%~6.3%. The majority (75%) of the Bacillus sp. in the RABC process was Bacillus subtilis which is known to enhance coagulation and precipitation by producing EPS. Hence, an adoption of a RABC process might be able to eliminate the disinfection unit process from a wastewater treatment system.

Construction and Characterization of the Vibrio parahaemolyticus Collagenase Inactivated Mutant (Vibrio parahaemolyticus collagenase 불활성화 돌연변이체의 제조 및 특성)

  • 이재원;전인준;강호영;차재호
    • Journal of Life Science
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    • v.14 no.2
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    • pp.362-367
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    • 2004
  • For better understanding of the host infection mechanism of Vibrio, a Vibrio parahaemolyticus collagenase mutant was generated by insertional inactivation of a vppC gene encoding extracellular collagenase. A recombinant DNA containing vppC::nptII was cloned into a suicide plasmid pDMS197, resulted in pVCM03. The recombinant suicide plasmid pVCM03 contained in E. coli $\chi$7213 was transferred to a wild-type V. parahaemolyticus 04 through conjugation. The recombinant vppC::nptII DNA in pVCM03 was exchanged with wild-type allele by homologous recombination resulting vppC mutant, V. parahaemolyticus CM. The mutant was selected and screened on TCBS media containing 10% sucrose and kanamycin. The mutation by allele exchange was confirmed with the comparison of the size of DNAs amplified by PCR. V. parahaemolyticus CM showed at least 4-fold less collagen-degrading activity than those of wild-type, and the mutant exhibited less cytotoxicity than that of wild-type in MTT assay.

An aluminum-based reflective nanolens array that enhances the effectiveness of a continuous-flow ultraviolet disinfection system for livestock water

  • Changhoon Chai;Jinhyung Park
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.258-270
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    • 2023
  • Climate change has worsened droughts and floods, and created conditions more likely to lead to pathogen contamination of surface water and groundwater. Thus, there is a growing need to disinfect livestock water. Ultraviolet (UV) irradiation is widely accepted as an appropriate method for disinfecting livestock water, as it does not produce hazardous chemical compounds and kills pathogens. However, UV-based disinfection inevitably consumes electricity, so it is necessary to improve UV disinfection effectiveness. Aluminum-based reflective nanolens arrays that enhanced the effectiveness of a continuous-flow UV water disinfection system were developed using electrochemical and chemical processes, including electropolishing and two-step anodization. A continuous UV disinfection system was custom designed and the parts were produced using a three-dimensional printer. Electropolished aluminum was anodized at 40 and 80 V in 0.3 M oxalic acid, at 120 and 160 V in 1.0 M phosphoric acid, and at 200 and 240 V in 1.5 M citric acid. The average nanolens diameters (D) of the aluminum-based reflective nanolens arrays prepared using 40, 80, 120, 160, 200, and 240 V anodization were 95.44, 160.98, 226.64, 309.90, 296.32, and 339.68 nm, respectively. Simple UV reflection behind irradiated water disinfected Escherichia coli O157:H7 in water more than did the non-reflective control. UV reflection and focusing behind irradiated water using an aluminum-based reflective nanolens array disinfected E. coli O157:H7 more than did simple UV reflection. Such enhancement of the UV disinfection effectiveness was significantly effective when a nanolens array with D 226.64 nm, close to the wavelength of the irradiated UV (254 nm), was used.

Effect of Scutellariae Radix as a Novel Antibacterial Herb on the ppk(Polyphosphate Kinase) Mutant of Salmonella typhimurium

  • Hahm, Dae-Hyun;Yeom, Mi-Jung;H.Lee, Eun-Joo;Shim, In-Sop;Lee, Hye-Jung;Kim, Hong-Yeoul
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1061-1065
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    • 2001
  • The antibacterial effects of water extracts of Scutellariate Radix (a dried root of Scutellaria baicalensis GEORGI) and its major flavonoid components, Baicalin and Baicalein, on Salmonella typhimurium, a representative enteric pathogen, were studied. Through a Kriby-Bauer disc analysis, the growth-inhibition activity of Scutellariae Radix against. S. typhimurium was found to be compatible with commercial antibiotics, such as ampicillin, chloramphenicol, and streptomycin. In contrast, the growth of a nonpathogenic E. coli strain was unaffercted by Scutellariae Radix. To examine the effect of polyphosphate kinase (ppk), a putative virulence factor, on the antibacterial activity of Scutellariae Radix, the growth profile of a ppk mutant of S. typhimurium was investigated in a tryptic soy broth containing different concentrations of water extracts of Scutellariae Radix. The ppk mutant was able to grow in 6 mg/ml of water extracts of Scutellariae Radix, whereas in 6 mg/ml of water extracts of Scutellariae Radix, whereas the wild-type could not, implying that the inactivation of ppk made S. typhimurium more resistant to the antibacterial activity of Scutellariae Radix. No enhanced resistance was observed in a ppk mutant of S. typhimurium complemented with a ppk expression vector. The attenuation of the virulence by ppk inactivation was also observed in a virulence assay using BLAB/c mice. Neither Baicalin nor Baicalein exhibited any growth-inhibition activity against S. typhimurium. The water extracts of Scutellariae Radix stimulated the transcription of ppk, especially in the early growth-stage of S. typhimurium.

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Inactivation of Microorganisms and Browning Enzymes in Angelica keiskei Juice Using High Hydrostatic Pressure (초고압을 이용한 신선초 녹즙의 살균 및 갈색화 효소의 불활성화)

  • Lee, Dong-Un;Park, Ji-Yong;Lee, Yun-Bom;Yeo, Ick-Hyun
    • Korean Journal of Food Science and Technology
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    • v.27 no.6
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    • pp.991-996
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    • 1995
  • Effects of high hydrostatic pressure on microorganisms and browning enzymes in Angelica keiskei juice were investigated using response surface methodology. The optimum process condition for maximum reduction of total aerobes was $5700\;kg_f/cm^2$ (558.6 MPa) pressure and 7.16 min process time, and 3.44 log cycle reduction of total aerobes was predicted at the optimum condition. E. coli, initially $8.8{\times}10^3\;CFU/ml$, was completely inactivated by high hydrostatic pressure at all process conditions ($3800{\sim}6700\;kg_f/cm^2\;pressure;\;3{\sim}17\;min\;process\;time$). Polyphenol oxidase and peroxidase were partly inactivated by the high hydrostatic pressure. It was also indicated that inactivation of microorganisms and browning enzymes by hydrostatic pressure is dependent on pressure rather than process time.

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Inactivation of Pathogenic Bacteria by Addition of Thermophilic Bacteria in the Thermophilic Aerobic Oxidation(TAO) System (고온호기산화장치의 고온미생물 첨가에 의한 병원성 미생물의 불활성화)

  • Lee W. I.;H. Tsujii;T. Maki;Lee M. G.
    • Journal of Animal Environmental Science
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    • v.10 no.2
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    • pp.111-118
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    • 2004
  • This study analyzed temperature increase, microorganism changes, and inactivation of pathogenic microorganisms in pig slurry when treated with thermophilic microorganisms in Thermophilic Aerobic Oxidation(TAO) system. An amount of $6 m^3$ of pig slurry was treated in an $18 m^3(3.0\times2.5\times2.4 m)$ reactor for 5 to 7 days in two groups: the control of pig slurry only and the treatment of pig slurry with 6 liters of thermophilic microorganism(Bacillus sp.). To study the microorganism changes in the reactor, the populations of aerobic mesophilic microorganisms, thermophilic microorganisms and general pathogens were analyzed. To study the inactivation of pathogenic microorganisms, the levels of E. coli, Salmonella sp, Crytosporidium parvum and Giardia lamblia were analyzed. The temperature inside the reactor ranged from 18 to $62^{\circ}C$ for the control while far the treatment group it ranged from 18 to $66^{\circ}C$, showing a slightly higher array. With regard to changes in microorganisms, both mesophilic and thermophilic organisms decreased from $3.1\times10^6$ to $1.2\times10^2$ CFU/ml and from $1.0\times10^4$ to $8.0\times10^1$ CFU/ml, respectively, in the control. In the treatment, on the other hand, mesophilic organisms decreased from $3.0\times10^8$ CFU/ml to $8.6\times10^5$ CFU/ml while thermophilic organisms increased sharply from $2.0\times10^6$ to $1.2\times10^8$ CFU/ml. For pathogens, Salmonella and Giardia were not detected either before or after the treatment, while E. coli and C. parvum were found to be $10^5$ CFU/ml each before treatment and negative after it. From this experiment, it was concluded that thermophilic microorganisms could effectively sanitize liquid compost by generating high temperature in the TAO system, which in turn would inhibit the growth of pathogenic organisms.

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