• 한국동물위생학회지
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• 제19권1호
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• pp.46-54
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• 1996

This study was perfomed to examine the distribution of causative agent of piglets diarrhea in eastern Chonnam province from February 1994 to March 1995. The causative agents of diarrhea were examined by bacterial culture test, parasitological test and serological test against PED, TGE and Rota. The 35 isolated E. coli were tested for antibiotic sensitivity. The results obtained were as follows. 1. The incidence of 81 piglets with diarrhea was most prevalent as 39.5% in the age of 2 to 4 weeks, compare to 34.6% in the age of 5 to 8 weeks and 16.0% under 1 weeks and 9.9 % in the age of 9 to 11 weeks after birth. 2. The incidence of 81 piglets diarrhea showed bacterial diarrhea(75.3%), viral diarrhea (35.8%) and parasitological diarrhea(18.5%). When compared the isolation frequency of each etiological agent, enteropathogenic E. coli was most prevalent as 55.5% in bacterial diarrhea, rotavlrus enteritis as 18.5% in viral diarrhea and trichuriasis as 13.6% in parasitological diarrhea. 3. The complicated infection of piglets was most prevalent as 41.7% in rotavirus enteritis with enteropathogenic E coli in 24 complicated piglets diarrhea. 4. In antibiotic sensitivity test, isolates showed moderatly resistance to Tobramycin, Amikacin, Imipenem, Trimethoprim/Sulfamethoxazole, Gentamicin, Ampicillin but sensitivty to Ticarcillin/K. The 30 E. coli isolate showed multiple drug resistances in 3 different antibiotics.

• Journal of Microbiology and Biotechnology
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• 제27권12호
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• pp.2141-2150
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• 2017

This study was conducted to develop a screening method using Colilert-18 and a device for the detection of E. coli from agri-food production environments and fresh vegetables. The specificity and sensitivity of Colilert-18 by temperature ($37^{\circ}C$ and $44^{\circ}C$) were evaluated with 38 E. coli and 78 non-E. coli strains. The false-positive rate was 3.8% (3/78) and 0% (0/78) at $37^{\circ}C$ and $44^{\circ}C$, respectively. The detection limit of E. coli at $37^{\circ}C$ at <1.0 log CFU/250 ml was lower than that at $44^{\circ}C$. The efficiency of the developed device, which comprised an incubator equipped with a UV lamp to detect E. coli in the field, was evaluated by measuring the temperature and UV lamp brightness. The difference between the set temperature and actual temperature of the developed device was about $1.0^{\circ}C$. When applying the developed method and device to various samples, including utensils, gloves, irrigation water, seeds, and vegetables, there were no differences in detection rates of E. coli compared with the Korean Food Code method. For sanitary disposal of culture samples after experiments, the sterilization effect of sodium dichloroisocyanurate (NaDCC) tablets was assessed for use as a substitute for an autoclave. The addition of one tablet of NaDCC per 50 ml was sufficient to kill E. coli cultured in Colilert-18. These results show that the developed protocol and device can efficiently detect E. coli from agri-food production environments and vegetables.

• 미생물학회지
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• 제18권3호
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• pp.148-152
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• 1980

The microwave of 2450 MHz, generated by a household cooking oven, was evaluated for its applicability to melt various rehydrated media and to remove dissolved oxygen from tubed media for anaerobic culture. The effect on the sterilization of E. coli in selective media was also evaluated. The following results were obtained. 10 The microwave oven was useful in saving time for melting media and in eliminating heat and combustion gas from the laboratory, which were inevitable by-products in the conventional flame method. 2) Dissolved oxygen could be removed without boiling over by exposing the tubes of anaerobic culture medium after putting them in a wire basket in a beaker with water. 30 The count of E. coli during the melting of MacConkey and EMB agar were similar to those treated with open flame. The microwave treatment was not considered a possible mean to replace autoclaving even in these selective media.

• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.72-77
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• 2009

Here, we demonstrate that the overexpression of the GroELS chaperone system, which assists the folding of intracellular proteins and prevents aggregation of its biological targets, can enhance the thermotolerance of Escherichia coli strains and facilitate the production of recombinant protein under thermal stress. The overexpression of GroELS led to an about 2-fold higher growth rate of E. coli XL-1 blue than control at $45^{\circ}C$ and induced the growth of the strain even at $50^{\circ}C$, although the growth was not sustained in the second-round culture. The effect of GroELS overexpression was also effective on other E. coli strains such as JM109, $DH5{\alpha}$, and BL21. Finally, we have shown that coexpression of GroELS allows us to produce recombinant protein even at $50^{\circ}C$, a temperature at which the protein production based on E. coli is not efficient. This study indicates that the employment of the GroELS overexpression system can expand the range of environmental conditions for E. coli.

• 한국환경보건학회지
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• 제22권2호
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• pp.10-18
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• 1996

Experiments were carried out to find the possibility of an economic production of single cell protein(SCP) in mixed culture by Cellulomonas sp. KL-6 and a second organism. The second organism, strain LI-10, was isolated from the large intestines of a mouse. 1. When these strains were mixed, cell growth and carboxymethyl cellulase (CMCase) activity were increased to about 63% and 161%, respectively compared with that of single culture of strain KL-6. We found the mixed culture as a proper method of degradation of cellulose in our study. 2. Strain LI-10 was identified as E. coli. 3. This strain produced trace amounts of cellobiose, but glucose was not found in detectable amounts in the filter paper(FP) medium. 4. $CaCO_3$ injected in the medium at the ratio of 0.1% not only enhanced cell growth but also was effective as an acid neutralizing agent. 5. When this organism was cultured under the optimal medium (glucose 0.1%, $NH_4Cl$ 0.1%, yeast extract 2.0%, $KH_2PO_4$ 0.1%, KCl 0.05%, pH 7.2 and a temperature 30$\circ$C) for 5 days, a cell mass produced 1.18 g/l. The results showed the increase of cell mass up to 300% compared to 0.28 g/l produced in CMC medium.

• 한국미생물·생명공학회지
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• 제26권1호
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• pp.15-22
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• 1998

일반적으로 Streptomyces류는 성장이 늦고 유지가 어려운 반면, E. coli는 배양기간이 짧고 유전자 조작도 간편한 장점이 있기 때문에, E. coli를 이용하여 유용단백질을 생산하는 연구가 일반적인 흐름이다. 그러나 E. coli에서 외래유전자를 도입하여 대량으로 생산을 시키는 경우에 비용해성의 inclusion body를 형성하는 경우가 많으므로 용해성의 활성형 단백질을 생산하기 위하여는 여러 가지 조건을 고려하여야 한다. 본 논문에서는 aklavlnone 11-hydroxylase gene(dnrF)을 E. coli BL2l에서 발현시킬 때의 배양조건을 배양온도와 IPTG농도의 두가지 요소를 조합하여 변형시키는 방법으로, 활성형 단백질의 생산을 최대화하고 inclusion body의 형성을 최소화하는 배양조건을 조사하였다. 그 결과, 37$^{\circ}C$에서 배양했을 때에는 0.02mM의 IPTG를 첨가하였을 때 inclusion body를 가장 적게 만들고, 그에 따라 생산되는 효소활성도 가장 높았다. 반면, 28$^{\circ}C$로 배양온도를 낮추었을 때에는 0.06mM의 IPTG를 첨가하였을 때 aklavinone 11-hydroxylase효소가 최대로 생산됨을 SDS-PAGE 및 효소 활성측정으로 확인하였다. IPTG농도를 0.1 mM로 높인 경우에는 28$^{\circ}C$, 37$^{\circ}C$에서 모두 aklavinone 11-hydroxylase효소가 과발현되어 Inclusion body를 가장 많이 생성하였음을 알 수 있었다. 방선균에서 동일 유전자를 대량발현시키는 경우 발현된 단백질의 효소 활성은 있으나 SDS-PAGE상에서의 단백질의 관찰이 불가능하였고, 동시에 단백질의 정제시 효소활성이 소실되어 정제가 불가능하였다. 이러한 효소 활성의 소실의 원인은 세포내의 어떤 저분자물질일 것으로 추정되며 본 연구에서 제작한 antibody를 이용하면, 효소의 정제가 용이하게 수행될 것이다. 특히 대장균계에서의 활성형 효소의 최적발현조건에서 세포를 배양하거나, inclusion body의 refolding을 실시한 후, 항체를 이용한 Western blot assay를 지표로 효소를 정제하면, 미지의 cofactor의 정체도 밝혀지고 aklavinone 11-hydroxylase류의 효소 특성 연구에 큰 도움이 될 것이다. 또한 이 효소를 이용한 다양한 종류의 bioconversion을 실시하여 그동안 background 때문에 생성된 product의 검출이 불가능했었던 소량의 생성산물의 분석도 가능할 것으로 판단되어 금후의 bioconversion연구에 기대하는 바가 크다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.731-734
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• 2001

An efficient fermentation strategy for the high level production of ultra-high-molecular weight poly(3-hdyroxybutyrate) (PHB) was developed. Although the cell and PHA concentrations obtained by flask cultures at different initial pH (6.0 or 6.9) were almost same level, the molecular mass of PHB produced were quite different along with the initial pH. When a recombinant Escherichia coli XL1-Blue harboring pJC2 containing the Alcaligenes latus PHB biosynthesis genes was cultivated in flask culture (pH 6.0), the PHB having a very high molecular weight of 22 MDa could be produced while only below 1 MDa at initial pH 6.9. The ultra-high-molecular weight PHB could be synthesized to high concentration of 89.8 g/L resulting in the PHB productivity of 2.07 g/L-h by simple fed-batch culture. In this study, we report that PHB having various molecular mass can be produced by employing metabolically engineered E. coli strains harboring the plasmids of different copy numbers containing the A. latus phbCAB genes.

• 미생물학회지
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• 제39권4호
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• pp.293-295
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• 2003

Total 70 isolates of Escherichia coli obtained from pigs were studied. Forty four isolates had $\textregistered$-hemolytic activity which was heat labile. Minimum inhibitory concentration test indicated that 40 isolates (57.1%), 15 isolates (21.4%), 23 isolates (32.9%), and 5 isolates (7.1%) were resistant to ampicillin, cephalothin, gentamicin, and norfloxacin, respectively. None of them were extended spectrum $\textregistered$-lactamases (ESBLs) producer when the double disk synergy test (DDST) was performed.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권6호
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• pp.371-374
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• 2002

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)［P(3HB/V)］, by fed-batch culture of recombinant Escherichia coli harboring a plasmid containing the Alcaligenes latus polyhy-droxyalkanoate (PHA) biosynthesis genes, was examined in two pilot-scale fermentors with air supply only, In a 30 L fermentor having a XLa value of 0.11 S­$^1$, the final P(3HB/V) concentration and the P(3HB/V) content obtained were 29.6 g/L and 70.1 wt%, respectively giving a productivity of 1.37 g P(3HB/V)/L-h. In a 300 L fermentor having a XLa of 0.03 S­$^1$, the P(3HB/V) concentration and the P(3HB/V) content were 20.4 g/L and 69 wt%, respectively giving a productivity of 1.06g P(3HB/V)/L-h. These results suggest that economical production of P(3HB/V) is possible by fed-batch culture of recombinant E. coli in a large-scale fermentor having low KLa value.

• 미생물학회지
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• 제34권1_2호
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• pp.20-25
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• 1998

PCBs를 분해하는 Pseudomonas sp. P20 균주의 pcbC gene이 재조합 된 E. coli CK1092 균주를 이용하여 2,3-dihydroxybiphenyl dioxygenase 생산에 미치는 배양조건을 검토하였다. E. coli CK1092를 2% sucrose가 포함된 LB 배지를 기본배지로 하여 질소원, 금속이온 등의 영향을 조사한 결과, $Fe^{3+}$와 $Fe^{2+}$이 $10^{-5}M$의 농도일때 효소생산이 증가되었으며 배양최적온도는 $37^{\circ}C$, 배양초기 pH 7.0일때 효소생산이 우수하였다. 배양조에서의 배양조건은 초기 pH 7.0, 통기량 1 v/v/m, 교반속도 200rpm에서 가장 효소생성이 우수하였다.