• 제목/요약/키워드: E. coli culture

검색결과 531건 처리시간 0.026초

재조합 내열성 트레할로스 합성효소의 생산 (Production of Recombinant Trehalose Synthase from Thermus caldophilus GK24)

  • 최재열;차월석;신현재
    • KSBB Journal
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    • 제21권4호
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    • pp.298-301
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    • 2006
  • 트레할로스 합성효소(trehalose synthase)의 효율적인 생산을 위하여, 5 종류의 plasmid를 형질전환 시킨 재조합 E. coli를 이용하여 균체생산량과 효소발현량을 비교하였다. Trehalose synthase의 활성은 fusion partner를 이용한 system 에서는 활성이 나타나지 않았으며, IPTG 유도 발현 시스템보다 항시적 발현 시스템을 사용하는 E. coli K12/pHCETS에서 가장 높은 활성을 나타내었다. 선별된 재조합 E. coli K12/pHCETS를 사용하여 회분식 및 유가배양을 수행하였으며, 유가식 배양의 경우 균체논도는 20 g/L, 최종 trehalose synthase 활성은 13.7 U/ml을 나타내었다. 이러한 결과는 트레할로스 생산을 위한 trehalose synthase가 재조합 E. coli의 발효에 의해 경제적으로 생산되어질 수 있다는 가능성을 보여 주었다.

내열성장독소 생산 대장균의 판정 (Assay of Heat Stable Enterotoxin Producing E. coli)

  • 장우현;김문교;최명식;양남웅;고광욱;서정기
    • 대한미생물학회지
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    • 제18권1호
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    • pp.53-58
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    • 1983
  • Enterotoxigenic E. coli is one of causative agents of the infantile diarrhea and traveler's diarrhea. A modified infant mouse assay(IMA) was developed for the detection of heat stable enterotoxin (ST) of E. coli isolated from diarrheal and control infants and assay system was established with using enterotoxin producing reference strains. The supernatant of the 24 hour-shaking culture of E. coli in Casamino Acid Yeast Extract Salt Broth(CYES-2) was ingested orally into the 2-4 day old ICR mice. After the mice were kept at $25^{\circ}C$ for 4 hours, they were sacrificed and the gut weight body weight ratio(GW/BW) was taken as the index of fluid accumulation induced by heat stable enterotoxin of E. coli. The results obtained were as follows; 1. The GW/BW responses of IMA tested with enterotoxin reference strains of E. coli(E. coli O148H28:$ST^+LT^+$, E. coli $O78H^-:ST^+LT^+$, E. coli O15H11:$ST^-LT^+$, E. coli O1H7:$ST^-LT^-$) appeared ta be ST dose-dependent, and not LT-dependent. From the dose-response curve, $25{\mu}l$ of culture supernatant was determined as test amount of the IMA. 2. Frequency distribution of IMA result from 643 strain of E. coli showed normal distribution at low GW/BW ratio and dispersed pattern at high GW/BW ratio. The GW/BW ratios of $0.056{\pm}0.004(mean{\pm}SD)$ of normal distribution which distributed from 0.044 to 0.068(P<0.01) was considered as ST negative. Thus the GW/BW ratio above 0.069 could be regarded as ST positive.

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구기자(Lycii fructus) 추출물이 미생물 생육에 미치는 영향 (The Influence of Lycii fructus Extracts on the Growth and Physiology of Microorganism)

  • 주인선;성창근;오만진;김찬조
    • 한국식품영양과학회지
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    • 제26권4호
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    • pp.625-631
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    • 1997
  • 구기자 추출물이 S. cerevisiae D7l과 L. casei KC-TC3165, 그리고 E. coli DH5균의 생육에 미치는 영향을 검토하였다. 그 결과 구기자 추출물은 S. cerevisiae D7l과 L. casei KCTC 3165의 생육을 증가시켰으며 E. coli DH5에 대하여는 약간의 생육저해가 있었다. 구기자methanol 추출물을 첨가하여 미생물의 생육을 측정하였을 때 L. casei KCTC 3165와 S. cerevisiae D7l는 각각 1.0%와 1.5% 첨가구에서 가장 높은 생육 촉진 효과를 보였다. 즉 구기자 추출물은 대장균의 생육은 저해하였으나, 인체에 유익한 균으로 알려진 젖산균에 대하여는 생육을 촉진하는 특성이 있었다. 한편, E. coli DH5는 구기자즙 10%, 그리고 methanol, chloroform 추출물 각각 1.0%첨가시 형태적 이상을 일으켜 길이가 8배 이상이 길어졌다. 구기자의 methanol추출물 중 회분성분이 E. coli DH5균에 이상현상의 원인이 되었음을 확인 하였다. 또한 이들 중 $Na^{+}$, $K^{+}$$Na^{+}$$K^{+}$ 혼합물이 0.135% 이상에서 균의 형태가 길어지는 이상 현상이 있었다.

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Evaluating Commercial Spray Applications of Lactic Acid, Hot Water, and Acidified Sodium Chlorite for the Reduction of Escherichia coli on Beef Carcasses

  • Kang, Dong-Hyun;Lee, Sun-Young
    • Food Quality and Culture
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    • 제2권1호
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    • pp.55-60
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    • 2008
  • This study examined the effects of lactic acid spray, hot water spray, or their combined treatment, as well as the effects of acidified sodium chlorite (ASC), for the decontamination of Escherichia coli on beef carcass surfaces using a commercial intervention system. With this system, the effects of 2 or 4% lactic acid (v/v), hot water ($89{\pm}1^{\circ}C$), or their combined treatment, were examined in terms of reducing inoculated E. coli. ASC (266 ppm), which was adjusted to pH 2.5 using acetic acid or citric acid, was applied using a hand-held spray system. When the beef carcasses were treated with 2 or 4% lactic acid for 10.4 s, less than 1 log reductions of inoculated E. coli were observed. A hot water spray treatment for 9.8 s resulted in a 2.1 log reduction of inoculated E. coli. However, when the hot water was followed with either 2 or 4% lactic acid, no difference in E. coli reduction was found between the hot water alone or the combined treatment with lactic acid. When ASC was adjusted to pH 2.5 with acetic acid and citric acid, 3.8 and 4.1 log reductions of E. coli were observed, respectively. Overall, the lactic acid spray treatment was least effective, and the ASC treatment was most effective, for the E. coli decontamination of beef carcasses. Therefore, these data suggest that ASC would be a more effective intervention against E. coli than most of the methods currently being used. However, more research is required to evaluate the effects of ASC on other organisms, as well as to identify application methods that will not affect meat quality.

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유기산의 저해를 조절하기 위한 Escherichia coli의 전기투석배양 (Control of Organic Acid Inhibition in Escherichia coli Culture with Eledtroudialysis)

  • 김인호;윤태호
    • 한국미생물·생명공학회지
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    • 제23권6호
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    • pp.747-754
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    • 1995
  • E. coli growth is inhibited by organic acids produced in the broth. In order to reduce the inhibition, an electrodialysis unit was used. Model solutions (acetic acid plus distilled water or M-9 medium) were tested in the unit for investigating the optimum condition of current and voltage. Electrodialysis cultures were performed with the optimum condition where the highest current efficiency could be attained. The distilled water plus acetic acid gave us a higher current efficiency than the M-9 plus acetic acid. Electrodialysis efficiently removed acetic acid and so enhanced the specific growth rate of E. coli compared with the control experiment without clectrodialysis.

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Detection of Escherichia coli O157:H7 Using Combined Procedure of Immunomagnetic Separation and Test Strip Liposome Immunoassay

  • Kim, Myung-Hee;Oh, Se-Jong;Durst, Richard-A.
    • Journal of Microbiology and Biotechnology
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    • 제13권4호
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    • pp.509-516
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    • 2003
  • A model system for the immnunochemical detection of Escherichia coli O157:H7 using a combined immunomagnetic separation (IMS) and test-strip liposome immunoassay (LIA) procedure was developed. Immunomagnetic beads coated with anti-E. coli O157 IgG antibodies were used to separate the E. coli O157 (including the H7 serotype) from culture. Immunoliposomes, whose surface was conjugated to goat anti-E. coli O157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used as a detection label. The test strip, onto which antibodies to goat IgG were immobilized, was the immunosensor capturing immunoliposomes that did not bind to E. coli O157:H7 on the immunomagnetic bead-E. coli O157:H7 complexes. In experiments, pure cell culture suspensions of $10^5 E.$ coli O157:H7 organisms per ml produced a measurable signal inhibition, whereas a weak yet detectable signal inhibition occurred with $10^3CFU/ml$. The inhibition signals increased, when the incubation time for IMS was extended to 90 min and higher IgG-tag density (0.4mol%) was used on the liposomes. With 0.2 and 0.4mol% IgG-tagged liposomes, the IMS-LIA procedure showed more improved signal inhibitions than those of a direct (no IMS) LIA. The combined assay, which measures the instantaneous signal from immunoliposomes, can be completed within 90 min, making it significantly faster than conventional plating methods and enzyme-linked immunosorbent assay (ELISA). Accordingly, it is quite feasible to use the combined immunoassay format of IMS and dye-loaded immunoliposomes for the detection of E. coli O157:H7.

대장균의 실관투석배양 (Hollow Fiber Dialysis Culture of E. coli)

  • 김인호;윤태호
    • KSBB Journal
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    • 제9권5호
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    • pp.492-498
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    • 1994
  • 설관막 장치를 이용하여 대장균의 투석배양을 수 행했다. 투석액으로 배지 내의 초산을 제거함으로써 초산의 저해효과를 경감할 수 있었다. 초산 생성속도는 포도당과 용존산소농도에 대단히 민감하였고, 따라서 막을 통한 포도당의 투과속도는 산소공급속도와 균형을 유지해야 했다. 막을 통해 포도당이 천 천히 공급될 때, 대장균의 비성장속도는 포도당 투 과속도에 좌우되었고 초산의 생성은 억제되였다.

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미생물 용해가 가능한 Iysozyme 분비 균주의 분리 및 특성 (Isolation of lysozyme producing bacteria capable of solubilizing microbial cells)

  • 구오펭페이;서선근;장레이;김효상;오영기;장덕진
    • KSBB Journal
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    • 제23권3호
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    • pp.187-192
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    • 2008
  • Lysozyme-producing microorganisms were isolated to obtain bacteria which can efficiently solubilize microbial cells. Cells of normal and chloroform-treated Escherichia coli and Micrococcus Iysodeikticus were used as model substrates to isolate lysozyme-producing microorganisms and investigate the efficiency of cell lysis. The culture supernatant of the isolate New1 (98% similarity of 16S rDNA sequence with Thermomonas haemolytica) showed different lytic characteristics for different substrates. Thermal treatment (autoclave) of substrate cells showed a significant effect on cell solubilization by culture supernatant of the New1. For autoclaved substrate cells, E. coli, M. Iysodeikticus and chloroform-treated E. coli were solubilized by 58.7%, 49.4% and 79.1%, respectively, in the culture supernatant of New1. The lytic activity of New1 was mainly caused by lysozyme produced by the isolate. It was also showed that New1 exhibited high protease activity and a little cellulase activity.

Pilot 규모에서의 재조합 대장균을 이용한 (R)-3-Hydroxybutyric acid 생산 (Pilot Scale Production of (R)-3-Hydroxybutyric acid by Metabolically Engineered Escherichia coli.)

  • 최종일;이승환;최성준;이상엽
    • 한국미생물·생명공학회지
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    • 제32권3호
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    • pp.243-248
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    • 2004
  • 산업적 R3HB의 생산을 위한 재조합 대장균의 pilot규모에서의 유가식 배양과 연속식 배양을 연구하였다. Pilot 규모에서의 R3HB생산을 위하여 안전한 two plasmid system pBRRed와 pMCS 105를 제작하였으며, 제작된 plasmids을 이용하여 여러 다른 대장균을 형질 전환하였다. 얻어진 재조합 대장균들을 30 l의 발효기에서 회분식 배양한 결과 대장균 XL-10 Gold(pBRRed, pMCS105)가 가장 높은 R3HB 농도를 보였다 30 1 발효기에서 대장균 XL-10 Gold (pBRRed, PMCS105)을 유가식 배양한 결과 22.4 g/1의 R3HB가 얻어졌으며, 생산성은 0.97 g/1-h를 보였다. 고농도의 R3HB를 고생산성으로 얻기 위하여 유가식 배양으로 높은 균체 농도를 얻은 후 연속 배양으로 R3HB를 생산하는 전략을 개발하였다. 그 결과 0.2 $h^{-1}$ 의 dilution rate에서 R3HB 생산성은 5.06 g/1-h를 보였다. 이러한 결과는 산업적 규모에서 재조합 대장균을 이용하여 R3HB를 고농도, 고생산성으로 얻을 수 있다는 것을 보여준다.

가축에서 대장균 감염증 (Colibacillosis in domestic animals, a review)

  • 송희종;채효석
    • 한국동물위생학회지
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    • 제21권4호
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    • pp.413-429
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    • 1998
  • Escherichia coli is recovered from a wide variety of infections in many animals species. It may be a primary or secondary agent. Nursing and young animals are particularly susceptible, and urinary tract infections are frequent. The various serotypes of E coli are intestinal inhabitants of animals including humans and probably infect most mammals and birds : therefore, they have a cosmopolitan distribution. Colibacillosis refers to any totalized or systemic infection caused entirely or partly by E coli. Collibacillosis in mammals is most often a primary enteric disease, whereas collibacillosis in poultry is typically a secondary located or systemic disease occurring when host defenses have been impaired or overwhelmed. Other opportunistic bacteria, which can be identified by culture, may play a similar role to that of I coli in secondary infections. Collectively, infections caused by E coli are responsible for significant economic losses to the animal performance. From the standpoint of pathogenic mechanisms and diseases, four major categories of E coli are recognized : enterotoxigenic(ETEC), enteropathogenic (EPEC), enteroinvasive(EIEC), and enterohemorrhagic(EHEC). In addition, two less-well-defined E coli categories are recognized in animals and humans : enteroaggregative and cytotoxin necrotizing factor-positive. The aforementioned categories are represented by different serotypes. Certain serotypes show a host preference and are encountered more frequently in some disease syndromes. Of the four major categories, ETEC is the most common cause of diarrhea in calves, lambs, and pigs. Strains in the other categories cause the less-common diarrhea and other disease syndromes. Enterotoxins and pilus antigens are the two most prominent virulence factors thus far identified for ETEC. Two enterotoxins, one heat-stable(ST) and one heat-labile(LT), are produced by enterotoxigenic strains of E coli : not all culture produce both of these plasmid-based enterotoxins.

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