• 공업화학
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• 제19권5호
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• pp.491-496
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• 2008

해양 어류인 Mugil cephalus로부터 에폭사이드 가수분해효소(epoxide hydrolase, EH) 유전자를 PCR을 이용하여 클로닝하고, pColdI 및 pET-21b(+) 발현벡터에 삽입시켜 재조합 Escherichia coli 생촉매를 개발하여 각각에 대하여 가수분해 활성을 비교하였다. 재조합 E. coli 생촉매 $10mg\;dcw\;mL^{-1}$을 사용하여 20 mM 라세믹 styrene oxide를 입체선택적 가수분해를 한 결과, (R)-styrene oxide 기질에 대해서 입체선택성을 보였다. pET-21b(+)를 발현벡터로 사용하여 M. cephalus의 EH 유전자를 저온 발현시킨 재조합 E. coli 생촉매를 사용하여, 약 40 min 반응을 통해 광학순도 99% ee (enantiomeric excess) 이상인 (S)-styrene oxide를 최종 수율 24.5% (이론수율 50%)로 제조할 수 있었다.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2009년도 제38회 동계학술대회 초록집
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• pp.47-47
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• 2010

Discharge under the water is very hard and demand considerable high voltage. But specially improved electrode can generate plasma discharge to salty water with relatively low voltage. A round shape ceramic electrode having many pinholes combined with metallic one can generate plasma. 400 volt, 10 kHz and 3 micro second pulse width were applied to repeatedly running synthetic seawater with 10 L/m velocity, containing cultivated E. coli and Bacillus. As a result, 18, 94, 99.97, 100, 100 % disinfection rates to E. coli and 17.1, 17.1, 82.9, 99.4, 99.9 % disinfection rates to Bacillus subtilis were achieved to 1, 2, 3, 4, 5 times repetitive treatment respectively. In the plasma condition, the ions and electrons are separated and new kinds of components are re-synthesized by the intensive movement of the components. Especially chlorine ions are separated and recombined to residual free chlorine like HOCl, $OCl^-$. The residual free chlorine concentrations of discharged water were 0.25, 0.88, 1.39, 1.59, 1.66 mg $Cl_2$/L after 5 times treatment respectively. Another unconfirmed radical and oxidants for example, OH, $H_2O_2$, and $O_3$ can have an effect on microorganism of course.

• 한국임상수의학회지
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• 제29권5호
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• pp.377-383
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• 2012

송아지 설사병은 국내 축산산업에 큰 피해를 주는 중요한 질병이다. 다양한 감염성 원인체들이 송아지 설사병에 관련될 수가 있기 때문에 효과적인 치료를 위해서는 신속한 감별진단이 필수적이다. 따라서 소설사병 바이러스(BVDV), 소 코로나바이러스(BCoV), A형 소 로타바이러스(BRV), 살모넬라, $K99^+$ 대장균, Cryptosporidium parvum등의 6개의 주요 병원체들을 동시에 검출하는 두 개의 multiplex real-time PCR으로 구성된 PCR 패널을 개발하여 국내 농가에서 전북대학교 동물질병진단센터로 접수된 97개의 설사 분변을 검사하였다. 또한 현미경 검사법을 이용하여 97개의 분변에서 Coccidium 충란을 검사하였다. 개발된 multiplex PCR의 민감도는 BVDV, BCoV와 BRV의 경우는 0.1 $TCID_{50}$, $K99^+$ 대장균은 5 CFU, Salmonella는 0.5 CFU, Cryptosporidium는 50 oocysts로 측정되었다. 또한 multiplex PCR의 증폭효율은 병원체에 따라0.97에서 0.99였다. 국내에서 접수된97개의 분변 중 36개의 분변은 multiplex PCR에 의해 최소 하나의 병원체에 대해 양성으로 판정되었고, 6개의 샘플은 2개의 병원체에 동시에 양성반응을 보였다. 또한 1달 이상 연령의 송아지 분변48개에서는 Coccidium 충란이 발견되었다. 결과적으로, 새로이 개발된 multiplex real-time PCR은 BVDV, BCoV, BRV, $K99^+$ 대장균, Salmonella와 Cryptosporidium과 관련된 송아지 설사병을 신속하고 정확하게 진단할 수 있는 유용한 검사법이 될 수 있을 것으로 기대되며 향후 Coccidium까지 검출할 수 있는 동시 진단법이 개발될 필요가 있을 것으로 생각된다.

• 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.

• BMB Reports
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• 제32권4호
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• pp.363-369
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• 1999

The orf8(chlD) gene cloned from Streptomyces antibioticus T$\"{u}$99 was overexpressed using an E. coli system to confirm its biological function. Induction of the E. coli strain transformed with recombinant plasmid pRFJ 1031 containing orf8 resulted in the production of a 43,000 dalton protein. Glucose-1-phosphate thymidylyltransferase activity of the cell extract obtained from the transformed strain was 4-5 times higher than that of the control strain. The expressed protein was purified 18-fold from E. coli cell lysate using three chromatographic steps with a 17% overall recovery to near homogeneity. The N-terminal amino acid sequence of the purified protein agrees with the nucleotide sequence predicted from the orf8 gene. The SDS-PAGE estimated subunit mass of 43,000 dalton agrees well with that calculated from the amino acid composition deduced from the nucleotide sequence of the orf8 gene (43,000 Da). Also, the native enzyme has a monomeric structure with a molecular mass of 43,000 dalton. The purified protein showed glucose-1-phosphate thymidylyltransferase activity catalyzing a reversible bimolecular group transfer reaction, and was highly specific for dTTP and ${\alpha}$-D-glucose 1-phosphate as substrates in the forward reaction, and for dTDP-D-glucose and pyrophosphate in the reverse reaction.

• 대한수의학회지
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• 제26권1호
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• pp.69-77
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• 1986

The purpose of this study was the examination for presence of K99 antigen (K99), enterotoxigenicity, 0-groups, colicin and antibiotic susceptibility among E. coil isolated from calves and cows. A total of 49(18.7%) among 262 strains, isolated from 30(26.5%) out of 113 calves and cows, possesed K99, and thirty three of 49 $K99^+$ strains produced ST. Of the strains of diarrheal calf origin which less than 15 days old, a high correlation was observed between enterotoxigenic ability and K99: 92.3% of the $K99^+$ strains produced heat stable enterotoxin(ST). In O group typing of 33 $ST^+$ strains, they belonged to O20(48.4%), O8(9.1%), O9(6.1%), O139(6.1%), O149(6.1%), O101(3.0%), O115(3.0%), except six which were untypable or autoagglutinable. Of 262 E. coil isolates, 30 strains(13.3%) produced colicin and K99 were detected in 6 strains. One hundred eighty eight strains(71.8%) of 262 E. coil isolates were resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, alone or in combination thereof. One hundred and fourteen(60.6%) out of 188 drug resistance strains carried R factor($R^+$) which were transferable to the recipients by conjugation. Sixty five $R^+$ strains(57.0%) carried thermo-sensitive R plasmid.

• 대한수의학회지
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• 제34권4호
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• pp.759-768
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• 1994

E coli strains isolated from pigs were investigated with respect to antimicrobial resistances and prevalence of R-plasmids. Also determined were properties of R-plasmids by plasmid conjugation, curing and southern hybridization using gene probes. All of 400 E coli strains were resistant to CL and SU, and 0.3% to 96.8% of the strains were resistant to most antimicrobials such as TC, PG, AM, SM, CP, GM, EM, NM, etc, while all strains were sensitive to AK. All strains were also multiply resistant to three to twelve antimicrobials. The resistances to PG, SM, TC, AM, CP, SU and ST were transferable and supposed to be mediated by R-plasmids which were opportunistic for transposition into chromosome. Plasmids bigger in size than chromosomal DNA were considered as R-plasmids and most plasmids in small size (<4Kb) proved as cryptic plasmids or nonconjugative R-plasmids. In a strain(No 99), AM resistant property was determined from both chromosomal DNA and R-plasmid DNA which is bigger in size than chromosome.

• 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.

• 미생물학회지
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• 제45권2호
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• pp.177-184
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• 2009

우수한 생균제를 개발하기 위하여 안전성이 알려진 유산균을 대상으로 인체의 분변으로부터 300여 균주를 분리하고 내산성, 내담즙성, 내열성, 항균력, 항암 및 항바이러스 효과를 가지는 균주들을 선발하여 생균제 특성을 나타내는지를 알아보기 위해 본 실험을 수행하였다. 인체에서 분리한 여러 균주 중 Miny-148 균주는 낮은 pH 및 높은 담즙산에 대한 내성, 열처리에 대한 열안정성을 지녀 기초적인 probiotic 특성을 가진 균주로 선발되어, Lactobacillus pentosus (99.9% 상동성)로 동정되었다. 항균력 실험에서 Escherichia coli O157:H7을 비롯한 Shigella flexneri, Bacillus anthracis, Staphylococcus aureus, E. coli, Vibrio cholerae, V. vulnificus, Salmonella typhimurium, 그리고 및 methicillin-resistant S. aureus (MRSA) 균주 8종 등 총 16종의 병원성세균을 억제하였다. 또한 Miny-148은 결장암 세포인 HT-29 cell을 억제하였을 뿐 아니라, transmissible gastroenteritis virus의 생육을 저해하여 세포변성 억제효과를 가진 우수한 probiotic 특성을 지닌 균주로 분석되었다.

• Journal of Microbiology and Biotechnology
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• 제9권2호
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• pp.206-212
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• 1999

The gene orf7(oxi III) was expressed using an E. coli system in anticipation that it would encode dTDP-glucose 4,6-dehydratase which is involved in the biosynthesis of the olivose moiety of chlorothricin produced from Streptomyces antibioticus Tu99. The solubility of the expressed protein increased up to 20% under optimal induction conditions. The expressed protein was purified from the E. coli BL 21(DE3) cell lysate by a 28.5-fold purification in two chromatography steps with a 38% recovery to near homogeneity. The molecular weight and N-terminal amino acid sequence of the purified protein correlated with the predicted mass and sequence deduced from the orf7 gene. The purified protein was a homodimer with a subunit relative molecular weight of 38,000 Dalton. The expressed protein was found to exhibit dTDP-glucose 4,6-dehydratase activity and be highly specific for dTDP-glucose as a substrate. The values of K'm and V'max for dTDP-glucose were 28 $\mu$M and 295 nmol $min^{-1} (mg protein)^{-1}$, respectively. dTTP and dTDP were strong inhibitors of this enzyme.$NAD^+$, the coenzyme for dTDP-glucose 4,6-dehydratase, was tightly bound to the expressed protein.