• Title/Summary/Keyword: E. coli culture

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Engineering of a Microbial Cell Factory for the Extracellular Production of Catalytically Active Phospholipase A2 of Streptomyces violaceoruber

  • Lee, Hyun-Jae;Cho, Ara;Hwang, Yeji;Park, Jin-Byung;Kim, Sun-Ki
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1244-1251
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    • 2020
  • Phospholipase A2 (PLA2) from Streptomyces violaceoruber is a lipolytic enzyme used in a wide range of industrial applications including production of lysolecithins and enzymatic degumming of edible oils. We have therefore investigated expression and secretion of PLA2 in two workhorse microbes, Pichia pastoris and Escherichia coli. The PLA2 was produced to an activity of 0.517 ± 0.012 U/ml in the culture broth of the recombinant P. pastoris. On the other hand, recombinant E. coli BL21 star (DE3), overexpressing the authentic PLA2 (P-PLA2), showed activity of 17.0 ± 1.3 U/ml in the intracellular fraction and 21.7 ± 0.7 U/ml in the culture broth. The extracellular PLA2 activity obtained with the recombinant E. coli system was 3.2-fold higher than the corresponding value reached in a previous study, which employed recombinant E. coli BL21 (DE3) overexpressing codon-optimized PLA2. Finally, we observed that the extracellular PLA2 from the recombinant E. coli P-PLA2 culture was able to hydrolyze 31.1 g/l of crude soybean lecithin, an industrial substrate, to a conversion yield of approximately 95%. The newly developed E. coli-based PLA2 expression system led to extracellular production of PLA2 to a productivity of 678 U/l·h, corresponding to 157-fold higher than that obtained with the P. pastoris-based system. This study will contribute to the extracellular production of a catalytically active PLA2.

Isolation of Escherichia coli O157 in Children with Diarrhea (소아설사 환아에서의 Escherichia coli O157 분리)

  • Song, Wonkeun;Kim, Hyoun Tae;Lee, Kyu Man;Cha, Jae Kook;Lee, Kon Hee
    • Pediatric Infection and Vaccine
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    • v.4 no.1
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    • pp.73-78
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    • 1997
  • Purpose : Escherichia coli O157 can produce diarrhea as well as hemorrhagic colitis and hemolytic uremic syndrome. In many parts of North America, E. coli O157 often is the second or third most commonly isolated enteric bacterial pathogens. Recently, intakes of fast food, including hamburgers have increased in Korea. Therefore, E. coli O157 infection in Korea are likely to be increased. Methods : Stool samples from 317 pediatric diarrheal patients were analyzed by culture on sorbitol-MacConkey agar. Sorbitol-negative colonies were teated by E. coli O157 latex agglutination test. Results : Of the 317 specimens, one (0.3%) were E. coli O157:NM that not produced Shiga toxin. The 7 year old male patient who had complained of abdominal pain, vomiting and non-bloody diarrhea for 2 days. The patient was improved for 2 days after admission. Conclusions 1 Only one (0.3%) of all fecal samples were isolated E. coli O157 that not produced Shiga toxin. Therefore, routine stool culture for the isolation of E. coli O157 was not likely to be neccessary so far.

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The Effects of Antibacterial Activity of Exopolysaccharide Isolated from Tibetan Mushroom Culture against Foodborne Pathogenic Bacteria: A Preliminary Study

  • Kim, Tae-Jin;Seo, Kun-Ho;Chon, Jung-Whan;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.2
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    • pp.68-77
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    • 2021
  • This study aimed to evaluate the effects of antibacterial activity of Tibetan mushroom exopolysaccharide against foodborne pathogenic bacteria Staphylococcus aureus 305, Listeria monocytogenes ATCC19114, Escherichia coli O157:H7 ATCC42894 and Escherichia coli O55. The yield of exopolysaccharide isolated from Tibetan mushroom culture was 620 mg/L. The antibacterial activity of exopolysaccharide against foodborne pathogenic bacteria exhibited 15 mm and 12 mm clear zone against S. aureus 305 and L. monocytogenes ATCC 19114, respectively. However, no clear zone was observed against E. coli O157:H7 ATCC 42894 and E. coli O55. In conclusion, exopolysaccharide isolated from Tibetan mushroom culture have the antibacterial activity only against Gram-positive foodborne pathogenic bacteria.

High Level Production of Supra Molecular Weight Poly(3-Hydroxybutyrate) by Metabolically Engineered Escherichia coli

  • Park, Jong-il;Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.196-200
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    • 2004
  • The supra molecular weight poly(〔R〕-3-hydroxybutyrate) (PH B), having a molecular weight greater than 2 million Da, has recently been found to possess improved mechanical properties compared with the normal molecular weight PHB, which has a molecular weight of less than 1 million Da. However, applications for this PHB have been hampered due to the difficulty of its production. Reported here, is the development of a new metabolically engineered Escherichia coli strain and its fermentation for high level production of supra molecular weight PHB. Recombinant E. coli strains, harboring plasm ids of different copy numbers containing the Alcaligenes latus PHB biosynthesis genes, were cultured and the molecular weights of the accumulated PHB were compared. When the recombinant E. coli XL1-Blue, harboring a medium-copy-number pJC2 containing the A. latus PHB biosynthesis genes, was cultivated by fed-batch culture at pH 6.0, supra molecular weight PHB could be produced at up to 89.8 g/L with a productivity of 2.07 g PHB/L-h. The molecular weight of PHB obtained under these conditions was as high as 22 MDa, exceeding by an order of magnitude the molecular weight of PHB typically produced in Ralstonia eutropha or recombinant E. coli.

Growth Inhibition of Enteropathogenic Escherichia coli $A_2$and Escherichia coli $G_7$ by the Organic Acid Producing Bacteria (유기산 생성균에 의한 병원성 Escherichia Coli $A_2$와 Escherichia Coli $G_7$의 생육억제)

  • 백영진;배형석
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.111-118
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    • 1988
  • The growth inhibition of enteropathogenic Escheriohia coli $A_2$and Escherichia coli G$_7$, causing the diarrhea in piglets, by the organic acid producing bacteria was studied in vitro. The metabolites of the organic acid bacteria, such as lactic acid, acetic acid inhibited the growth of E. coli $A_2$and E. coli G$_7$ in BL medium. The more the organic acid producing bacteria have ability to produce the organic acids, the higher these bacteria excelled the inhibitory efficacy against enteropathogenic E. coli. Among the strains examined, Lactobacillus casei Y and Streptococcus faecium C showed relatively strong growth inhibition against enteropathogenic E. coli.. When the organic acid producing bacteria and the enteropathogenic E. coli were incubated simultaneously in BL medium, bacteriostasis of E. coli was observed when the pH of BL culture was lowered to 5.0, and bacteriocidal effect was observed when the pH became Bess than 4.5, E. coli. $A_2$was more resistant to the organic acid bacteria than E. coli G$_7$.

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Combination Effects of Potassium Sorbate and Sodium Benzoate with sodium Chloride on the Growth Inhibition of Escherichia coli and Salmonella typhimurium (Escherichia coli 와 Salmonella typhimurium 의 생육억제에 미치는 식염과 Potassium Sorbate, Sodium Benzoate의 병용효과)

  • Cho, Nam-Sook;Yang, Yeo-Young;Choi, Eon-Ho
    • Korean Journal of Food Science and Technology
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    • v.18 no.4
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    • pp.249-254
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    • 1986
  • An experiment was performed to investigate the combined effect of preservatives and the synergistic effect of sodium chloride to them on the inhibition of bacterial growth. Escherichia coli and Salmonella typhimurium were cultured with or without shaking in liquid media (pH 6) of tryptone-glucose-yeast extract or tryptic soy broth which contained 0.1% potassium sorbate and/or 0.03% sodium benzoate, equivalent to half of the maximum permissible levels, respectively. The growth of E. coli was more inhibited with one or both of the two preservatives by shaking culture than by non-shaking culture. For S. typhimurium the single treatment of the preservatives did not show inhibitory effect whereas the combined treatment of them showed bacteriostatic effect in shaking culture and a prolongation of lag phase in non-shaking culture. Addition of 2% sodium chloride to either potassium sorbate or potassium sorbate plus sodium benzoate remarkably increased the growth inhibition of E. coli for non-shaking cultivation but no effect observed for shaking cultivation. S. typhimurium was more sensitive to the addition of sodium chloride than E. coli in both shaking and non-shaking culture to show lower viable cell counts than initial numbers.

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Transfer and Expression of E. coli LacZ Gene in Boving Embryos by Co-culturing with Retrovirus Vector-Producing Cells (Retrovirus Vector를 생산하는 세포와 공동배양된 소 수정란의 E. coli LacZ 유전자 전이와 발현)

  • 김태완;박세필
    • Korean Journal of Animal Reproduction
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    • v.19 no.2
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    • pp.89-93
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    • 1995
  • In this study was demonstrate that retrovirus-mediated gene transfer is one of the promising alternatives to the conventional pronuclear DNA microinjection approach, especially in transferring the exogenous genes into the boving embryos. By co-culturing of zona of zona-free one-cell stage embryos with the retrovirus-producing cells for 24 hours followed by 6 days of culture in virus-free medium, we could get morulae and blastocysts expressing the E. coli LacZ genes which were transferred by our retrovirus vector. The results obtained in this study are summarized as follows : 1. Addition of 5$\mu\textrm{g}$/ml of polybrene in the embryo and virus-producing cell co-culture medium did not affect development of zona-free one-cell embryo. 2. Compared with the intact embryos removal of zona at one-cell stage before co-culturing with the virus-producing cells for one day caused only slight decrease of embryo develpment. 3. Co-culture of 625 zona-free one-cell stage embryos with the virus-producing cells resulted in 65(10.4%) morulae or blastocysts, and 12.3%(8/65) of the morulae or blastocysts were E. coli LacZ positive.

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Evaluation of Epidemic Characteristics of Extended Spectrum β-Lactamase Producing Bacteria Isolated from Blood Cultures (혈액배양에서 분리된 Extended Spectrum β-Lactamase 생성균의 역학적 특성 조사)

  • Seo, Choong-Won;Kim, Sang-Ha;Hwang, Seock-Yeon;Kim, Young-Kwon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.9
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    • pp.2516-2522
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    • 2009
  • The data which analyze the results of blood cultures are crucial basic information of the empirical therapy for patients with infection since the patterns of the species of microorganism isolated from blood and the results of antibiotic susceptibility test vary depending upon patients general features. Especially, in case of ESBL-producing bacteria, there is a close relation with use of antibiotics. Therefore, we carried out the research with the results of blood culture and antibiotic. 1. Total 39,305 cases of blood culture samples were investigated and positive patients of 2,216 (20.0%) were found. Among those, there were 40 patients with ESBL positive, and blood culture positive samples were 4,798 (12.2%). ESBL positive bacteria were found in 86 samples (including double checked culture bacteria). 2. The majority of ESBL producing bacteria were E. coli, K. pneumoniae and K. oxitoca as ordering based on the number. 3. The research showed the results that there were more females than male with the bacterias, more E. coli in over 50 years old aged group than other bacterias, more K. pneumoniae and K.oxitoca in less 1 year old aged group than other bacterias and largest numbers of patients with 13 patients (32.5%) in Chungcheongnam-do province were found. 4. The most common ESBL producing bacteria were E. coli throughout 3 years, but K, pneumoniae and K. oxitoca were also fairly found. Interestingly, E. coli was highly found in over 50 years old patients.

Antibacterial Activity against Pathogenic Bacteria of Lactiplantibacillus argentratensis Isolated from Blueberries (블루베리로부터 분리된 Lactiplantibacillus argentratensis의 병원성균에 대한 항균활성)

  • Natsag Lkhagvasuren;Gil-Ha Kim;Batchimeg Namshir;Woan Sub Kim
    • Journal of Dairy Science and Biotechnology
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    • v.41 no.4
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    • pp.191-202
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    • 2023
  • In this study, lactic acid bacteria (LAB) was isolated from blueberries. The isolated LAB were rod-shaped and gram-positive, as shown using gram staining. In addition, the identified bacteria showed high homology to Lactiplantibacillus argentoratensis. The culture supernatant was isolated from L. argentoratensis and its antibacterial activity against the pathogenic bacteria Salmonella and Escherichia was analyzed. Culture supernatants of L. argentoratensis significantly inhibited the growth of Salmonella. Enteritidis NCCP 16947, Salmonella Typhimurium NCCP 16960, and Salmonella. Thompson NCCP 11704. Interestingly, the higher the concentration of the culture supernatant, the more significant was the antibacterial activity. Additionally, the culture supernatant of L. argentoratensis showed significant antibacterial activity against Escherichia strains. To determine whether the antibacterial substance is stable to heat and pH, the LAB culture supernatant was heat-treated under 65℃ for 30 min, 75℃ for 15 min, 85℃ for 10 min, and 100℃ for 5 min. Measurement of antibacterial activity against pathogenic strains by adding 5% of heat-treated culture medium showed the same antibacterial activity as before heat treatment. However, in a test where the pH of the culture supernatant was adjusted to 7.0 from 3.73, no antibacterial activity was observed.

Production of Poly(3-hydroxybutyrate) [P(3HB)] with High P(3HB) Content by Recombinant Escherichia coli Harboring the Alcaligenes latus P(3HB) Biosynthesis Genes and the E. coli ftsZ Gene

  • Choi, Jong-Il;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.722-725
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    • 1999
  • Filamentation-suppressed recombinant Escherichia coli strain harboring the Alcaligenes latus polyhydroxyalkanoate (PHA) biosynthesis genes and the E. coli ftsZ gene was constructed and cultivated for the production of poly(3-hydroxybutyrate) [P(3HB)] with high concentration and high content. By the pH-stat fed-batch culture of this recombinant E. coli strain XL1-Blue(pJC5), the final cell concentration and P(3HB) concentration obtained in 44.25h were 172.2g cell dry weight/l and 141.9g P(3HB)/l, respectively, resulting in productivity of 3.21g P(3HB)/l-h. More importantly, the P(3HB) content obtained was 82.4 wt %, which was significantly higher than that obtained with the recombinant E. coli harboring only the PHA biosynthesis genes.

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