• Title/Summary/Keyword: E. coli culture

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A Study on Growth Inhibition of Escherichia coli and Salmonella typhimurium by Lactic Acid Bacteria (유산균에 의한 Escherichia coli와 Salmonella typhimurium의 생육억제에 관한 연구)

  • Kim, E.A.;Baick, S.C.;Chung, W.H.
    • Journal of Animal Science and Technology
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    • v.44 no.4
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    • pp.491-498
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    • 2002
  • The inhibitory effect of lactobacilli and bifidobacteria on the growth of typical intestinal pathogens, Escherichia coli and Salmonella typhimurium was studied. The degree of inhibition was measured by well disc assay and turbidimetry method. The strains which showed the higher antimicrobial activity were L. acidophilus La-5, L. acidophilus NCFM, L. casei Lc-01 on the average by using two different methods. The associative cultures were performed with selected 3 lactobacilli and 2 enteropathogens E. coli and S. typhimurium, respectively. Inhibition of pathogen began at 9hr after culturing so that viable counts was decreased rapidly. After 30hr incubation, there were no viable pathogens from the mixed culture. Under this experimental condition, the antimicrobial activity of lactic acid bacteria was not due to pH alone and supposed to different to the strains.

Antibacterial Activity of Powdered Spice against Escherichia coli and Staphylococcus aureus (향신료 분말의 Esdcherichia coli 와 Staphylococcus aureus 에 대한 항균작용)

  • 김미림;최경호;박찬성
    • Food Science and Preservation
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    • v.7 no.1
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    • pp.124-131
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    • 2000
  • Antibacterial activities of powdered spices(garlic , ginger, cinnamon and clove) against pathogenic Escherichia coli )157:H7 and Staphyloccus auresus were investigated. Spice powder was added in was exponetial phase of each bacterial culture . Growth inhibition was determined by the absorbance at 660nm and morphological changes of the cells were observed by transmission electron microscope (TEM). Ginger powder has the highest antibacterial activity, following cinnamon , clove and garlic has the least activity.Growth of Escherichia coli O157:H7 and Staphyloccus aureus were completely inhibited within 5 hours after addition of 1 % of garlic , 0.3% of ginger or cinnamon , 0.5% of clove powder on the exponential phase of the cells. Spice untreated cells of E. coli and S. aureus, the cytoplasm was entirely surrounded by rigid cell wall and cell walls formed a smooth layer well attached to the plasma membrane. In the cells of E. coli and S. aureus treated with spice powder, cell wall and plasma membrane were lysed and severely damaged. E.coli cells growth in the presence of spice powder showed plammolysis, the loss of electron dense material, the formation of extra cellular blebs and cytoplasm burst out from the cell. S .sureus cells grown in the presence of spice powder showed swell of cell wall, the loss of electron dense material , coagulation of cell cytoplasm and formation of extra cellular blebs. Severely damaged cells of S. aureus lost whole cytoplasm and left as ghost of the cell. Spice powder stimulated autolyssi and induced cell death.

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Evaluation of Dry Rehydratable Film Method for Detection of Coliform Bacteria and Escherichia coli (대장균군과 대장균의 검출을 위한 건조필름 방법의 평가)

  • Park, Heon-Kuk
    • The Korean Journal of Food And Nutrition
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    • v.22 no.4
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    • pp.696-700
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    • 2009
  • The adaptability of dry rehydratable film for the qualitative evaluation of coliform bacteria and Escherichia coli was tested. In general, culture methods that employ lactose broth or desoxycholate lactose agar are used for qualitative tests of coliform bacteria. Culture using lactose broth showed a high detection yield and low selectivity when compared to the dry rehydratable film. However, culture methods that employ lactose broth required a long time(48 hrs) for qualitative tests of coliform bacteria and complicated procedures were required to prepare the medium. The detection of coliforms using desoxycholate lactose agar had a slightly higher selectivity than the dry rehydratable film method, but this difference was not statistically significant. However, the preparation of the desoxycholate lactose agar was complicated. EC broth for the detection of E. coli showed the highest detection yield and lowest selectivity; however, this method required complicated procedures for preparation of the medium as well. Overall, the dry rehydratable film had a slightly lower detection yield than the other methods. The detection yield of dry rehydratable film method was over 37.1% at a concentration of 1 cfu/$m{\ell}$. Additionally, the dry rehydratable film method showed high selectivity and did not require preparation. However, because the selectivity of the dry rehydratable film was high, it took a long time(36 hrs) to detect E. coli. Overall, these findings indicate that dry rehydratable film can be used for qualitative detection of coliforms and E. coli.

Combined analysis of transcriptome and proteome for high cell density cultivation of Escherichia coli

  • Yun, Seong-Ho;Han, Mi-Jeong;Im, Geun-Bae;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.845-848
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    • 2001
  • For understanding physiology and metabolism under various culture conditions, combined analysis of transcriptome and proteome is attractable way. We have manufactured DNA microarray containing 2,850 genes including all functionally known and putative ones. In this study, we report analysis of transcriptome and proteome during the high cell density culture of E. coli by using DNA microarray and 2-DE. Fed-batch fermentation of E. coli was carried out by exponential feeding of nutrients until the maximum cell density reached 74 g dry cell weight/L (g DCW/L). Changes in transcriptome and proteome during the HCDC are analyzed qualitatively and quantitatively to provide their physiological and metabolic meanings.

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Characterization of Heat-Stable Enterotoxin of Enterotoxigenic Escherichia coli eKT-53 (장독성 대장균 eKT-53균주의 내열성 장독소의 성질)

  • 도대홍;김교창;김도영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.6
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    • pp.621-628
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    • 1991
  • Heat-stable enterotoxin(ST) from enterotoxigenic E. coli eKT-53($ST^{+}\;LT^{-}$, transformant from isolate KM-7) that was produced in succinate salts medium. The culture supernatant(crude ST) was purifed by mulitpled steps and investigated some characterization of the ST. The heatstability of purified ST activity was completely lost by treating at $100^{\circ}C$ for 30minutes. ST activity was lost by treatment at pH 1 and 12 conditions, while the activity was not reduced by treatment at pH 2~10, and then the ${\alpha}-amylase$ and pepsin was not decreased activity but disulfide reducing agnets was lost the activity. The molecular weight of the purified ST was approximately 4,200, the isoelectric point was about 4.0.

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Optimization of recombinant E. coli fermentation through biological manipulation and engineering control

  • Kim, Jeong-Yoon
    • The Microorganisms and Industry
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    • v.19 no.4
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    • pp.14-26
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    • 1993
  • Optimizing protein production in recombinant E. coli strains involves manipulation of genetic and environmental factors. In designing a production system, attention must be paid to gene expression efficiency, culture conditions and bioreactor configuration. Although not much emphasis was given to the physiology of host strains in this review, an understanding of the relationship between the physiology of host cell growth and the overproduction of a cloned gene protein is of primary importance to the improvement of the recombinant fermentation processes. Sometimes it is desirable to make use of gene fusion systems, e.g. protein A, polypeptide, gutathione-S-transferase, or pneumococcal murein hydrolase fusion, to facilitate protein purification.

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Evaluation of Various Escherichia coli Strains for Enhanced Lycopene Production

  • Jun Ren;Junhao Shen;Thi Duc Thai;Min-gyun Kim;Seung Ho Lee;Wonseop Lim;Dokyun Na
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.973-979
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    • 2023
  • Lycopene is a carotenoid widely used as a food and feed supplement due to its antioxidant, anti-inflammatory, and anti-cancer functions. Various metabolic engineering strategies have been implemented for high lycopene production in Escherichia coli, and for this purpose it was essential to select and develop an E. coli strain with the highest potency. In this study, we evaluated 16 E. coli strains to determine the best lycopene production host by introducing a lycopene biosynthetic pathway (crtE, crtB, and crtI genes cloned from Deinococcus wulumuqiensis R12 and dxs, dxr, ispA, and idi genes cloned from E. coli). The 16 lycopene strain titers diverged from 0 to 0.141 g/l, with MG1655 demonstrating the highest titer (0.141 g/l), while the SURE and W strains expressed the lowest (0 g/l) in an LB medium. When a 2 × YTg medium replaced the MG1655 culture medium, the titer further escalated to 1.595 g/l. These results substantiate that strain selection is vital in metabolic engineering, and further, that MG1655 is a potent host for producing lycopene and other carotenoids with the same lycopene biosynthetic pathway.

Heterologous Production of Streptokinase in Secretory Form in Streptomyces lividans and in Nonsecretory Form in Escherichia coli

  • Kim,, Mi-Ran;Choeng, Yong-Hoon;Chi, Won-Jae;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.132-137
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    • 2010
  • The skc gene encoding streptokinase (SK) with a molecular mass of approximately 47.4 kDa was cloned from Streptococcus equisimilis ATCC 9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the promoter for sprT [Streptomyces griseus trypsin (SGT)] was used in the host S. lividans TK24, a 47.4-kDa protein was detected along with a smaller hydrolyzed protein (44 kDa), suggesting that posttranslational hydrolysis had occurred as has been reported in other expression systems. The casein/plasminogen plate assay revealed that the plasmid construct containing the SGT signal peptide was superior to that containing the SK signal peptide in terms of SK production. Maximal production of SK was calculated to be about 0.25 unit/ml of culture broth, a value that was five times higher than that obtained with other expression systems using ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(${\Delta}DE3$)pLys under the control of the T7 promoter, a relatively large amount of SK was expressed in soluble form without hydrolysis. SK activity in E. coli/pET28a-$T7_pSK_m$ was more than 2 units/ml of culture broth, even though about half of the expressed protein formed an inactive inclusion body.

Rapid Detection of Escherichia coli in Fresh Foods Using a Combination of Enrichment and PCR Analysis

  • Choi, Yukyung;Lee, Sujung;Lee, Heeyoung;Lee, Soomin;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Oh, Hyemin;Lee, Yewon;Kim, Yujin;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.38 no.4
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    • pp.829-834
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    • 2018
  • The objective of this study was to determine the minimum enrichment time for different types of food matrix (pork, beef, and fresh-cut lettuce) in an effort to improve Escherichia coli detection efficiency. Fresh pork (20 g), beef (20 g), and fresh-cut lettuce (20 g) were inoculated at 1, 2, and 3 Log CFU/g of Escherichia coli. Samples were enriched in filter bags for 3 or 5 h at $44.5^{\circ}C$, depending on sample type. E. coli cell counts in the samples were enriched in E. coli (EC) broth at 3 or 5 h. One milliliter of the enriched culture medium was used for DNA extraction, and PCR assays were performed using primers specific for uidA gene. To detect E. coli (uidA) in the samples, a 3-4 Log CFU/mL cell concentration was required. However, E. coli was detected at 1 Log CFU/g in fresh pork, beef, and fresh-cut lettuce after 5, 5, and 3-h enrichment, respectively. In conclusion, 5-h enrichment for fresh meats and 3-h enrichment for fresh-cut lettuce in EC broth at $44.5^{\circ}C$, and PCR analysis using uidA gene-specific primers were appropriate to detect E. coli rapidly in food samples.

Evaluation of Uricult Trio Test as a Rapid Screening of UTI in Children with Fever (소아 요로감염 진단에 있어 신속배양검사(Uricult Trio)의 유용성에 관한 연구)

  • Lee, Jin;Kim, Eun Jin;Lee, Taek Jin;Chang, Jin Keun;Cha, Sung-Ho
    • Pediatric Infection and Vaccine
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    • v.17 no.2
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    • pp.74-82
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    • 2010
  • Purpose : The Dipslide culture test is a rapid method for diagnosis of urinary tract infection (UTI). The aim of this study is to determine the diagnostic availability of a urine Dipslide test for evaluation of UTI in febrile children. Methods : Urine specimens from 151 febrile infants were inoculated by a routine blood agar urine culture method and the Dipslide test at the same time. Following incubation for 16-24 hours, the results of the Dipslide test were read at the next visit. Both results of Dipslide and those of routine culture were compared. Results : The mean age of subjects was 15${\pm}$10.6 months. There were 150 infants (99.3%) who had fever with a mean duration of 2.6${\pm}$2.6 days. Thirty two infants (21.2%) were diagnosed as having UTI. Sensitivity and specificity of Uricult Trio CLED medium were 59.4% and 84.8%, respectively. Sensitivity and specificity of Uricult Trio E. coli medium were 60.0% and 96.2%, respectively. The Pearson correlation coefficient between results of Uricult Trio CLED medium and urine culture was 0.438 (P=0.01). Correlation between results of Uricult Trio E. coli medium and urine culture was 0.617 (P=0.01). Conclusion : The Dipslide test requires only 16-24 hours with high specificity in terms of UTI caused by E. coli without the problems associated with specimen delay. This test seems to be helpful for exclusion of UTI in febrile infants and it may reduce unnecessary hospitalization and antibiotic use. However, further studies are required before the product can be recommended as a routine diagnostic tool.