• Title/Summary/Keyword: E. coli culture

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

  • Choi, Jae-Youl;Cha, Wol-Suk;Shin, Hyun-Jae
    • KSBB Journal
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    • v.21 no.4
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    • pp.298-301
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    • 2006
  • A gene(GeneBank AF 135796) coding for a trehalose synthase from Thermus caldophilus GK24 was cloned into Escherichia coli K12 using five vector systems. The constitutive expression system(pHCETS) which shows the highest trehalose synthase activity from flask culture of recombinant E. coli was selected for the production of trehalose from maltose. For the shake flask culture, the final dry cell weight was 0.9 g/L and the trehalose synthase activity was 25 U/mL. Fed-batch culture of recombinant E. coli harboring plasmid pHCETS which uses the glycerolas a carbon source was performed in jar fermentor: the dry cell weight of 20 g/L and the trehalose synthase activity of 13.7 U/mL were attained in 48 h.

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

  • Chang, Woo-Hyun;Kim, Moon-Gyo;Choi, Myung-Sik;Yang, Nam-Ung;Ko, Kwang-Wook;Seo, Jung-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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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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The Influence of Lycii fructus Extracts on the Growth and Physiology of Microorganism (구기자(Lycii fructus) 추출물이 미생물 생육에 미치는 영향)

  • 주인선;성창근;오만진;김찬조
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.625-631
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    • 1997
  • This study was peformed to investigate the effects of Lycii fructus extracts on the growth and physiology of S. cerevisae D7l, L. casei KCTC 3165, and E. coli DH5. When Lycii fructus was added into solid culture media, the growth of S. cerevisea D7l and L. casei KCTC 3165 was increased, whereas that of E. coli DH5 was somewhat decreased. The growth of S. cerevisiea D7l and L. casei KCTC 3165 were much promoted in the culture media including methanol extract of 1.0% and 1.5%, respectively. E. coli DH5 was changed its morphology not only in 10% Lycii fructus juice but also 1.0% methanol and chloroform extract of Lycii fructus. Its size in those growth condition was eight times longer than that of the normal E. coli DH5. It was elucidated that elongation phenomenan of E. coli DH5 was also appeared by adding 0.135% of Na$^{+}$, $K^{+}$, and the mixture of Na$^{+}$ and $K^{+}$.TEX> +/.

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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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    • v.2 no.1
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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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Control of Organic Acid Inhibition in Escherichia coli Culture with Eledtroudialysis (유기산의 저해를 조절하기 위한 Escherichia coli의 전기투석배양)

  • 김인호;윤태호
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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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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    • v.13 no.4
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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.

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

  • 김인호;윤태호
    • KSBB Journal
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    • v.9 no.5
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    • pp.492-498
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    • 1994
  • A hollow fiber device was utilized to perform a dialysis culture for E. coli. Acetic acid inhibition on the growth of E. coli was relieved by dialyzing the acid from broth into a dialysate reservoir. The rate of acetic acid formation was very sensitive to the concentration of glucose and dissolved oxygen. Therefore it was found that the glucose permeation rate should be balanced with the oxygen supply rate. Specific growth rate of E. coli was determined by the glucose permeation rate through membrane. Under a low permeation rate, acetic acid formation was depressed in accordance with high dissolved oxygen concentration as well as low glucose concentration.

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

  • Guo, Pengfei;Seo, Sun-Keun;Zhang, Lei;Kim, Hyo-Sang;Oh, Young-Khee;Jahng, Deok-Jin
    • KSBB Journal
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    • v.23 no.3
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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 Scale Production of (R)-3-Hydroxybutyric acid by Metabolically Engineered Escherichia coli. (Pilot 규모에서의 재조합 대장균을 이용한 (R)-3-Hydroxybutyric acid 생산)

  • 최종일;이승환;최성준;이상엽
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.243-248
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    • 2004
  • Production of (R)-3-hydroxybutyric acid (R3HB) by fed-batch culture and continuous culture of metabolically engineered Escherichia coli harboring Ralstonia eutropha PHB biosynthesis and depolymerase genes was examined in a 30 1 pilot-scale fermentor. A new stable two-plasmid system, pBRRed containing the R. eutropha PHB depolymerase gene and pMCS 105 containing the R. eutropha PHB biosynthesis genes, was developed. Among a variety of E. coli strains harboring plasmids, recombinant E. coli XL-10 Gold (pBRRed, pMCS105) was able to produce R3HB with the highest efficiency in a batch culture. By the fed-batch culture of recombinant E. coli XL-10 Gold(pBRRed, pMCS 105) in a 30 1 fer-mentor, the final R3HB concentration was 22.4 g/l giving a productivity of 0.97 g/l-h. To produce R3HB to a high concentration with high productivity, a new strategy of fed-batch culture followed by a continuous culture was investigated. The maximum productivity and R3HB concentration were 5.06 g/l-h and 25.3 g/l, respectively. These results show that economical production of R3HB is possible by recombinant E. coli in large scale.

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

  • 송희종;채효석
    • Korean Journal of Veterinary Service
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    • v.21 no.4
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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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