• 한국식품위생안전성학회지
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• 제20권1호
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• pp.48-52
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• 2005

본 연구는 시판되는 증자 녹차의 에탄을 추출물과 에탄을 추출물로부터 분획한 분획물로부터 최근 병원성 식중독균으로서 문제시되고 있는 E. coli O157:H7균에 대한 항균성을 검색하였다. 에탄을 추출물로부터 그 유효성분을 조사하기 위하여 핵산, 에틸아세테이트, 클로로포름 및 물로서 분획한 후 MIC를 검색한 결과 에틸아세테이트 분획물이 $500{\mu}g/ml$의 농도로 가장 강한 활성을 나타내었으며, 헥산 분획물은 활성이 없었다. 가장 강한 항균력을 나타낸 에틸아세테이트 분회물의 농도별 항균력 및 pH에 대한 영향을 조사한 결과 균종에 따라 차이는 있지만 3균주 모두 $250-2000{\mu}g/disk$농도에서 생육이 억제되었다. 또한 E. coli O157:H7균은 산성조건에서는 생육이 가능하지만 pH 10 이상의 강한 알칼리 조건에서는 생육이 크게 억제되었다. $35^{\circ}C$에서 E. coli O157:H7 933과 932균에 대한 에틸아세테이트 분획물의 농도별 생육저해 효과를 검색한 결과 두 균주 모두 $250{\mu}g/ml,\;500{\mu}g/ml$의 농도에서는 12시간 이후부터 생육하기 시작하였으나 $1000{\mu}g/ml$의 농도에서는 생육이 크게 저해되었다.

• Journal of Microbiology and Biotechnology
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• 제30권1호
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• pp.109-117
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• 2020

Cre recombinase is widely used to manipulate DNA sequences for both in vitro and in vivo research. Attachment of a trans-activator of transcription (TAT) sequence to Cre allows TAT-Cre to penetrate the cell membrane, and the addition of a nuclear localization signal (NLS) helps the enzyme to translocate into the nucleus. Since the yield of recombinant TAT-Cre is limited by formation of inclusion bodies, we hypothesized that the positively charged arginine-rich TAT sequence causes the inclusion body formation, whereas its neutralization by the addition of a negatively charged sequence improves solubility of the protein. To prove this, we neutralized the positively charged TAT sequence by proximally attaching a negatively charged poly-glutamate (E12) sequence. We found that the E12 tag improved the solubility and yield of E12-TAT-NLS-Cre (E12-TAT-Cre) compared with those of TAT-NLS-Cre (TAT-Cre) when expressed in E. coli. Furthermore, the growth of cells expressing E12-TAT-Cre was increased compared with that of the cells expressing TAT-Cre. Efficacy of the purified TAT-Cre was confirmed by a recombination test on a floxed plasmid in a cell-free system and 293 FT cells. Taken together, our results suggest that attachment of the E12 sequence to TAT-Cre improves its solubility during expression in E. coli (possibly by neutralizing the ionic-charge effects of the TAT sequence) and consequently increases the yield. This method can be applied to the production of transducible proteins for research and therapeutic purposes.

• Food Science and Biotechnology
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• 제17권1호
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• pp.191-194
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• 2008

The antagonistic activity of probiotic strains (Bifidobacterium animalis BB-12, Bifidobacterium bifidum A, Bifidobacterium longum B6, Lactobacillus acidophilus ADH, Lactobacillus paracasei ATCC 25598, and Lactobacillus rhamnosus GG) against nalidixic acid resistant ($NA^R$) Escherichia coli O157:H7 MF1847, E. coli O157:H7 H2439, E. coli O157:H7 ATCC 43894, and E. coli O157:H7 C7927 was investigated using the agar-overlay, well diffusion, and broth culture tests. L. paracasei ATCC 25598 was the most effective probiotic strain in terms of in vitro antagonistic activity against $NA^R$ E. coli O157:H7, followed by L. rhamnosus GG, B. longum B6, and L. acidophilus ADH. The use of selected probiotic strains could be an effective pre-harvest intervention strategy to reduce the risk of $NA^R$ E. coli O157:H7 by maintaining a balanced microflora in animals and might provide many potential benefits in lieu of using antimicrobials.

• Asian-Australasian Journal of Animal Sciences
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• 제22권2호
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• pp.238-245
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• 2009

Pure culture experiments were conducted to assess the bacteriostatic and bactericidal effects of phlorotannins (PT) isolated from Ascophyllum nodosum (brown seaweed) on Escherichia coli O157:H7. In Exp. 1, one non-O157:H7 strain (25922) and three strains of E. coli O157:H7 (3081, EDL933 and E318N) were cultured in M9 medium with PT included at 0 (control), 25, 50 or $100{\mu}g/ml$ (n = 3). Bacterial growth was monitored by $OD_{600}$ at 0, 4, 6, 12 and 24 h, and by dilution plating at 0, 4, 6 and 24 h. All strains were inhibited (p<0.001) by PT to varying degrees. At 50 or $100{\mu}g/ml$, PT prevented growth of all four strains. At $25{\mu}g\;PT/ml$, growth of 25922, 3081, E318N and EDL933 was inhibited for 6, 12 and 24 h, respectively, but 25922 and 3081 resumed growth by 12 and 24 h. Direct plating confirmed bactericidal effects of PT on all four strains at $100{\mu}g/ml$, and on EDL933 and E318N at $50{\mu}g/ml$. In Exp. 2, strains 25922 and 3081 were incubated with no tannins or with $50{\mu}g/ml$ of PT, purified condensed tannins (CT) from Quebracho (Schinopsis balansaei), or purified tannic acid from Rhus semialata (Anacardiaceae) as hydrolysable tannins (HT). Strain 3081 was unaffected by HT or CT, but was completely inhibited (p<0.001) by PT at 4, 6 and 24 h. Strain 25922 was unaffected by HT, slightly inhibited by CT, and almost eradicated by PT at 4 and 6 h. Transmission electron microscopy revealed tannin-mediated alterations to bacterial cell walls. Phlorotannins from A. nodosum exhibit growth-inhibiting and bactericidal effects in vitro against the strains of E. coli O157:H7 investigated. Anti-E. coli efficacy of A. nodosum PT is superior to that of terrestrial tannins purified from Quebracho and from Rhus semialata.

• 한국지역사회생활과학회지
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• 제18권2호
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• pp.293-300
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• 2007

10종의 한약재 MeOH 추출물과 EtOH 추출물의 E. coli에 대한 항균력을 조사한 결과 소목과 오배자의 MeOH 추출물과 EtOH 추출물은 5 mg/ml 수준에서 항균활성을 나타내었으며 목단피와 황금등은 10 mg/ml 이상에서 항균성을 보였다. 최소저해농도는 MeOH 추출물에서는 $1.4{\sim}8mg/ml$, EtOH 추출물에서는 $1.2{\sim}12mg/ml$ 농도에서 항균활성을 보였으며 분획층 중 EtOAc층에서 성장억제효과가 가장 높게 나타났다. 항균력이 우수한 것으로 확인된 소목과 오배자 추출물의 미생물 증식억제 효과를 조사하기 위해 증식배지에 각각의 추출물을 0, 100, 300 및 500 ppm의 농도로 첨가하여 균주의 증식을 조사 한 결과 배양 후 24시간에 소목 추출물 무첨가구의 $OD_{620}$값이 0.6인 반면 300ppm 이상의 추출물 첨가 시 $0.02{\sim}0.1$정도로 균증식이 현저히 억제되었고, 오배자 추출물 무첨가구의 $OD_{620}$값이 0.5인 반면 500ppm 이상의 추출물 첨가 시 $0.02{\sim}0.2$정도로 균증식이 현저히 억제되었다. 이상의 결과로 볼 때 질병을 일으키는 E. coli의 감염을 예방하거나 또는 치료할 수 있는 한약재 추출물의 개발 가능성을 확인할 수 있었다.

• 한국축산식품학회지
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• 제32권1호
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• pp.62-67
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• 2012

This study calculated kinetic parameters of Escherichia coli O157:H7 and developed a probabilistic model to estimate growth probabilities of E. coli O157:H7 on polyethylene cutting boards as a function of temperature and time. The surfaces of polyethylene coupons ($3{\times}5$ cm) were inoculated with E. coli O157:H7 NCCP11142 at 4 Log $CFU/cm^2$. The coupons were stored at 13 to $35^{\circ}C$ for 12 h, and cell counts of E. coli O157:H7 were enumerated on McConkey II with sorbitol agar every 2 h. Kinetic parameters (maximum specific growth rate, Log $CFU/cm^2/h$; lag phase duration, h; lower asymptote, Log $CFU/cm^2$; upper asymptote, Log $CFU/cm^2$) were calculated with the modified Gompertz model. Of 56 combinations (temperature${\times}$time), the combinations that had ${\geq}$0.5 Log $CFU/cm^2$ of bacterial growth were designated with the value of 1, and the combinations that had increases of <0.5 Log $CFU/cm^2$ were given the value 0. These growth response data were fitted to the logistic regression to develop the model predicting probabilities of E. coli O157:H7 growth. Specific growth rate and growth data showed that E. coli O157:H7 cells were grown at $28-35^{\circ}C$, but there were no obvious growth of the pathogen below $25^{\circ}C$. Moreover, the developed probabilistic model showed acceptable performance to calculate growth probability of E. coli O157:H7. Therefore, the results should be useful in determining upper limits of working temperature and time, inhibiting E. coli O157:H7 growth on polyethylene cutting board.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2060-2065
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• 2016

Characteristics of E. coli O157:H7-specific infection bacteriophages (O157 coliphages) and broad-host-range bacteriophages for other E. coli serotypes (broad-host coliphages) were compared. The burst sizes of the two groups ranged from 40 to 176 PFU/infected cell. Distributions of the virulence factors stx1, stx2, ehxA, and saa between the two groups were not differentiated. Broad-host-range coliphages showed lower stability at $70^{\circ}C$, in relation to O157 coliphages. However, O157 coliphages showed high acid and ethanol tolerance by reduction of only 22% and 11% phages, respectively, under pH 3 and 70% ethanol for 1 h exposure. Therefore, these results revealed that the O157 coliphages might be more stable under harsh environments, which might explain their effective infection of the acid-tolerant E. coli O157:H7.

• 생명과학회지
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• 제21권12호
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• pp.1710-1715
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• 2011

Nano plasma ion (NPi) generator에서 발생한 NPi의 미생물에 대한 살균 효과를 측정하기 위해 미생물 종류, 조사 시간, 챔버 용적, 이온 수량, 거리에 따라 실험 하였다. 먼저 6종의 미생물 Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella pneumoniae, Staphylococcus aureus, Bacillus subtilis를 대상으로 실험한 결과 미생물 종류에 따라 각각 다른 감소율을 나타냈으며, 그람 음성균인 E. coli가 96.57%로 가장 높았고, 그람 양성균 중 포자를 생성하는 B. subtilis가 57.41%로 가장 낮았다. 그리고 NPi 조사시간에 따라 살균력 측정한 결과, 반응 초기에 대부분의 미생물이 사멸하였으며 이후 서서히 증가하였다. 또한 챔버의 크기에 따른 E. coli의 감소율을 비교하였으며 $0.005m^3$부터 $30m^3$까지 5개 챔버에서 NPi를 2시간 조사한 결과 용적이 증가함에 따라 포화이온 농도는 낮아졌고 이와 함께 살균력도 감소하였다. 이에 $1m^3$ 챔버에 NPi generator를 추가로 설치하여 포화 이온농도에 따른 E. coli의 감소율을 알아보았고 포화 이온 농도가 증가함에 따라 감소율도 함께 증가하는 것으로 나타났다. 마지막으로 NPi generator의 거리에 따른 E. coli의 감소율을 확인하였고 이온이 직접적으로 분출되는 부분의 99.19%를 제외한 나머지 위치에서 팬에 의한 이온 순환으로 포화농도가 비슷하게 유지 되었으며 약 97%의 감소율을 나타냈다.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.593-598
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• 2004

The arsenical resistance (ars) operon was cloned from a 67-kilobase pair (kb) plasmid, which was previously shown to be responsible for arsenic salts resistance in K. oxytoca D12. When plasmid pAE48, carrying the ars operon, was transformed into E. coli, transformed cells displayed enhanced survival in the presence of 4 mM arsenite, 50 mM arsenate, or 0.4 mM antimonite. The nucleotide sequence of the 5.6-kb fragment encoding arsenical resistance revealed five open reading frames (ORFs), which were predicted to encode polypeptides of 12.8 (arsR), 13.4 (arsD), 62.6 (arsA), 45 (arsB), and 16.7 (arse) kilodaltons (kDa). Each ORF was preceded by a ribosome binding site. A putative promoter-like sequence was identified upstream of arsR, and a possible termination site was found downstream of arsC. When the deduced amino acid sequences of the K. oxytoca Dl2 Ars proteins were compared with the amino acid sequences of the E. coli R773 Ars proteins, a significant amino acid similarity was observed (87.9％ for ArsR, 89.2％ for ArsD, 83.2％ for ArsA, 92.6％ for ArsB, and 91.3％ for ArsC), suggesting an evolutionary relationship of the ars genes of E. coli plasmid R773 and K. oxytoca Dl2.

• Environmental Engineering Research
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• 제12권2호
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• pp.64-71
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• 2007

The necessity of disinfecting playground soil is an important issue, because pathogenic protozoa, bacteria, and parasite eggs remain viable for several months and can infect children. UV irradiation has been used to decontaminate water but its effectiveness on soil is unclear. We determined the efficacy of UV radiation for inactivation of an indicator bacteria, E. coli (strain ATCC 8739), on playground soil. While 99% inactivation of E. coli in the soil was readily achieved by UV radiation within 55 min at $0.4\;mW\;cm^{-2}$, complete inactivation was not achieved, even after prolonged treatment at $4\;mW\;cm^{-2}$. This was attributed to the irregular surface of the soil. A small number of E. coli escaped the UV radiation because they were situated in indentations or under small particles on the soil surface. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) confirmed that the surface characteristics of the soil is the major limiting factor in the inactivation of E. coli by UV radiation. Thus UV treatment may not be adequate for disinfecting some soils and should be carefully evaluated before being used on playground soils.