• 제목/요약/키워드: E. coli concentration

검색결과 818건 처리시간 0.023초

Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli

  • Park, Eun-Hee;Lee, Won-Heong;Jang, Mi-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • 제25권10호
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    • pp.1709-1713
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    • 2015
  • Sepiapterin is a precursor for the synthesis of tetrahydrobiopterin (BH4), which is a wellknown cofactor for aromatic amino acid hydroxylation and nitric oxide synthesis in higher mammals. In this study, a recombinant Escherichia coli BL21(DE3) strain harboring cyanobacterial guanosine 5’-triphosphate cyclohydrolase 1 (GCH1) and human 6-pyruvoyltetrahydropterin synthase (PTPS) genes was constructed to produce sepiapterin. The optimum conditions for T7 promoter–driven expression of GCH1 and PTPS were 30℃ and 0.1 mM isopropyl-β-D-thioglucopyranoside (IPTG). The maximum sepiapterin concentration of 88.1 ± 2.4 mg/l was obtained in a batch cultivation of the recombinant E. coli, corresponding to an 18-fold increase in sepiapterin production compared with the control condition (37℃ and 1 mM IPTG).

Enhanced Production of ${\varepsilon}$-Caprolactone by Coexpression of Bacterial Hemoglobin Gene in Recombinant Escherichia coli Expressing Cyclohexanone Monooxygenase Gene

  • Lee, Won-Heong;Park, Eun-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • 제24권12호
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    • pp.1685-1689
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    • 2014
  • Baeyer-Villiger (BV) oxidation of cyclohexanone to ${\varepsilon}$-caprolactone in a microbial system expressing cyclohexanone monooxygenase (CHMO) can be influenced by not only the efficient regeneration of NADPH but also a sufficient supply of oxygen. In this study, the bacterial hemoglobin gene from Vitreoscilla stercoraria (vhb) was introduced into the recombinant Escherichia coli expressing CHMO to investigate the effects of an oxygen-carrying protein on microbial BV oxidation of cyclohexanone. Coexpression of Vhb allowed the recombinant E. coli strain to produce a maximum ${\varepsilon}$-caprolactone concentration of 15.7 g/l in a fed-batch BV oxidation of cyclohexanone, which corresponded to a 43% improvement compared with the control strain expressing CHMO only under the same conditions.

Activity of Chlorelaa vulgaris Associated by Escherichia coli W3110 on Removal of Total Organic Carbon in Continuous River Water Flow System

  • Kong, Surk-Key;Nakajima Toshiuki
    • ALGAE
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    • 제17권3호
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    • pp.195-199
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    • 2002
  • We investigated the association of Chlorella vulgaris and E. coli W9110 in removal of total organic carbon with the lab-scaled continuous river water flow system (CRWFS). Artificial wastewater was applied at two levels of organic carbon concentration; 1,335 $mg{\cdot}l^{-1}$ in the treatment (T)-1 and 267 $mg{\cdot}l^{-1}$ in T-2. The highest densities of C. vulgaris were $8.3{\times10^6\;cells{\cdot}ml^{-1}$ in T-1 and $6.9{\times}10^6\;cells{\cdot}ml^{-1}$ in T-2. The maximum densities of E. coli W3110 were $2.0{\times}10^8$ clony forming unit (CFU)${\cdot}ml^{-1}$ in T-1 and $3.9{\times}10^8\;CFU{\cdot}ml^{-1}$ in T-2. The densities increased during the first 11 days in T-q and 4 days in T-2, and decreased rapidly till 35th day, then increased slightly afterwards. This trend was prominent in T-2. It was inplied that wider range of nutrients was required in the growth of heterotrophic bacteria in T-2 than in T-1. The algal biomass should be increased effectively for the successful removal of organic carbon.

복분자 에탄올 추출물의 식중독 관련 위해 세균에 대한 항균활성 분석 (Antimicrobial Activity of the Ethanol Extract from Rubus coreanum against Microorganisms Related with Foodborne Illness)

  • 전연희;순샤오칭;김미라
    • 한국식품조리과학회지
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    • 제28권1호
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    • pp.9-15
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    • 2012
  • This study analyzed the antibacterial activity of a Rubus coreanum (Bokbunja) ethanol extract. The antimicrobial activity was determined by disc diffusion, minimum inhibitory concentration (MIC), and growth inhibition methods with seven kinds of bacteria related to foodborne illness (Bacillus cereus, Escherichia coli, Escherichia coli O157:H7, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhimurium). In the results, disc diffusions of the ethanol extract from R. coreanum (9.8-17.5 mm at $4,000{\mu}g/disc$) clearly showed the antimicrobial activity of the extract against all tested microorganisms. Rubus coreanum promoted an inhibitory effect as follows: E. coli O157:H7 > P. aeruginosa > L. monocytogenes > E. coli > S. aureus > B. cereus ${\geq}$ S. typhimurium. In the MIC test, R. coreanum showed high antimicrobial effect against L. monocytogenes at 500 ppm. Moreover, the R. coreanum ethanol extract showed strong growth inhibition against microorganisms, similar to the MIC results. These results show that a R. coreanum ethanol extract has powerful antimicrobial activity against all tested microorganisms, suggesting that R. coreanum will be useful as a potential natural preservative.

Sterilization of Bacteria, Yeast, and Bacterial Endospores by Atmospheric-Pressure Cold Plasma using Helium and Oxygen

  • Lee Kye-Nam;Paek Kwang-Hyun;Ju Won-Tae;Lee Yeon-Hee
    • Journal of Microbiology
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    • 제44권3호
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    • pp.269-275
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    • 2006
  • Atmospheric-pressure cold plasma (APCP) using helium/oxygen was developed and tested as a suitable sterilization method in a clinical environment. The sterilizing effect of this method is not due to UV light, which is known to be the major sterilization factor of APCP, but instead results from the action of reactive oxygen radicals. Escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae deposited on a nitrocellulose filter membrane or Bacillus subtilis spores deposited on polypropylene plates were exposed to helium/oxygen plasma generated with AC input power at 10 kHz, 6 kV. After Plasma treatment, nitrocellulose filter membranes were overlaid on fresh solid media and CFUs were counted after incubation overnight. D-values were 18 sec for E. coli, 19 sec for S. aureus, 1 min 55 sec for S. cerevisiae, and 14 min for B. subtilis spores. D-values of bacteria and yeast were dependent on the initial inoculation concentration, while the D-value of B. subtilis spores showed no correlation. When treated cells were observed with a scanning electron microscope, E. coli was more heavily damaged than S. aureus, S. cevevisiae exhibited peeling, and B. subtilis spores exhibited shrunken morphology. Results showed that APCP using helium/oxygen has many advantages as a sterilization method, especially in a clinical environment with conditions such as stable temperature, unlimited sample size, and no harmful gas production.

Rv3168 Phosphotransferase Activity Mediates Kanamycin Resistance in Mycobacterium tuberculosis

  • Ahn, Jae-Woo;Kim, Kyung-Jin
    • Journal of Microbiology and Biotechnology
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    • 제23권11호
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    • pp.1529-1535
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    • 2013
  • Tuberculosis is a worldwide epidemic disease caused by Mycobacterium tuberculosis, with an estimated one-third of the human population currently affected. Treatment of this disease with aminoglycoside antibiotics has become less effective owing to antibiotic resistance. Recent determination of the crystal structure of the M. tuberculosis Rv3168 protein suggests a structure similar to that of Enterococcus faecalis APH(3')-IIIa, and that this protein may be an aminoglycoside phosphotransferase. To determine whether Rv3168 confers antibiotic resistance against kanamycin, we performed dose-response antibiotic resistance experiments using kanamycin. Expression of the Rv3168 protein in Escherichia coli conferred antibiotic resistance against $100{\mu}M$ kanamycin, a concentration that effected cell growth arrest in the parental E. coli strain and an E. coli strain expressing the $Rv3168^{D249A}$ mutant, in which the catalytic Asp249 residue was mutated to alanine. Furthermore, we detected phosphotransferase activity of Rv3168 against kanamycin as a substrate. Moreover, docking simulation of kanamycin into the Rv3168 structure suggests that kanamycin fits well into the substrate binding pocket of the protein, and that the phosphorylation-hydroxyl-group of kanamycin was located at a position similar to that in E. faecalis APH(3')-IIIa. On the basis of these results, we suggest that the Rv3168 mediates kanamycin resistance in M. tuberculosis, likely through phosphotransferase targeting of kanamycin.

Salmonella choleraesuis의 증균(增菌)에 관한 연구 -Tetrathionate Broth Base에 대한 옥도용액(沃度溶液)의 함량(含量)이 증균효과(增菌效果)에 미치는 영향- (Enrichment of Salmonella choleraesuis in Tetrathionate Broth -Effect of Concentration of Iodine Solution on the Growth of Sal. choleraesuis-)

  • 정길택;탁연빈
    • 대한수의학회지
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    • 제16권1호
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    • pp.53-57
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    • 1976
  • The growth of Sal. choleraesuis and its kunzendorf variety in tetrathionate broth containing various amounts of iodine solution was studied and compared with that of Sal. typhi, Sal. typhimurium and E. coli. The results obtained were as followings. 1. When 2.0 ml of iodine solution, normal amount, was added to 100 ml of tetrathionate broth base, the number of Sal. choleraesuis decreased rapidly until 24 hours after inoculation and slightly increased 48 hours after inoculation. The numbers of Sal. choleraesuis var. kunzendorf and E. coli decreased rapidly and none of the organisms recovered 24 and 48 hours after inoculation, respectively. The growth of Sal. typhi and Sal. typhimurium, however, was not inhibited at all. 2. When 4.0 ml of iodine solution was added, to 100 ml of tetrathionate broth base, the growth of all the organisms was inhibited, among which Sal. choleraesuis, Sal. choleraesuis var. kunzendorf, and E. coli were not recovered 24 and 48 hours after inoculation. 3. When reduced amounts of iodine solution, 1.0 ml and 0.5 ml, were added to 100 ml of tetrathionate broth base, the growth of all the organisms was not inhibited.

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Bacterial Die-Off in Continuous River Water Flow System

  • Kong, Surk-Key;Toshiuki Nakajima
    • 한국환경과학회지
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    • 제12권8호
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    • pp.847-852
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    • 2003
  • It was examined carefully that the bacterial die-off between Chlorella vulgaris and E. coli. W3110 was tested through adding TOC (total organic carbon) with the lab-scaled continuous river water flow system (CRWFS). Artificial synthetic wastewater was applied at two levels of organic carbon concentration; 1,335 mg/l in treatment type 1 and 267 mg/l in type 2. In both types, the population densities of Chlorella vulgaris were similar in a maximum 8.25 ${\times}$ 10$\^$6/ cells/ml (type 1) and 6.925 ${\times}$ 10$\^$6/ cells/ml (type 2). The maximum densities of E. coli. W3110 were 2.0 ${\times}$ 10$\^$8/ colony forming unit (CFU)/ml in type 1 and 3.9 ${\times}$ 10$\^$8/ CFU/ml in type 2. The densities increased for 11 days in type 1 and 4 days in type 2, then decreased rapidly till the 35th day, then slightly increased again. This trend was prominent in type 2. It implied that a wider range of nutrients was required in the growth of heterotrophic bacteria in type 2 than in type 1. We could not expect successful bacterial die-off if the wastewater retention time was not furnished sufficiently.

화분발효물을 함유한 Gelidium corneum - 젤라틴 혼합필름의 항균활성과 돈육의 포장재 적용 (Antimicrobial Activity of a Gelidium corneum - Gelatin Blend Film Containing Fermented Pollen Extract and Its Application in the Packaging of Pork Loins)

  • 홍윤희;임금옥;송경빈
    • 한국축산식품학회지
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    • 제29권3호
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    • pp.391-395
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    • 2009
  • 화분발효물(FPE)을 함유하는 Gelidium corneum - 젤라틴(GCG) 필름을 제조하여 돈육의 포장재로 사용하였다. FPE를 첨가한 GCG필름의 투습계수는 대조구 필름 보다 향상되었고 FPE 첨가 농도가 증가함에 따라 Escherichia coli O157:H7와 Listeria monocytogenes에 대한 항균효과는 증가하였다. 특히, 0.15% FPE 첨가는 E. coli O157:H7와 L. monocytogenes에 대해 대조구 대비 각각 2.98 and 3.68 Log CFU/g 감소시켰다. 또한 돈육 시료를 E. coli O157:H7 와 L. monocytogenes로 접종시킨 후 GCG 필름으로 포장한 후 저장 중 미생물 수의 변화를 측정한 결과, 0.15% 화분발효물이 첨가한 GCG 필름으로 포장 시 E. coli O157:H7와 L. monocytogenes 숫자가 저장 4일 후 대조구 대비 각각 1.49, 1.01 Log CFU/g 더 감소시켰다. 이러한 연구 결과는 0.15% 화분발효물이 첨가한 GCG 필름으로 포장한 돈육의 유통기한이 증대될 수 있음을 시사한다.

回春凉膈散과 龍石散이 抗炎作用에 미치는 影響 (Experimental study on the Anti-inflammatory and wound healing effect of Hoichunyanggyuksan and Yongseksa.)

  • 강승원;노석선
    • 한방안이비인후피부과학회지
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    • 제12권1호
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    • pp.47-78
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    • 1999
  • Hoichunyonggyuksan(HCYGS) and Yongseksan(YSS) are important prescriptions that have been used in oriental medicine for stomatitis and wound healing. The study was done to evaluate the inhibitory effects of cytotoxicity, formation of superoxide on the macrophage and neutrophil, prostaglandins($PGE_2$), interleukins($IL-1{\Beta}$), collagenase activity and synthesis of collagen and DNA. The results were obtained as fo1lows: 1. HCYGS and YSS were not showed the proliferation difference of human fibroblast and monocyte in all concentrations to be experimented and in result, it was concluded that they have no cytotoxicity. 2. YSS inhibited the formation of superoxide to $48\% at the concentration of $0.001\%$ in the mouse monocyte. 3. HCYGS inhibited the formation of superoxide to $13\%$ at the concentration of $0.001\%$ as compared with control in the human monocyte. 4. HCYGS and YSS inhibited the formation of superoxide to $25\%\;and\;35\%$ respectively at the concentration of $0.0001\%\;and\;HCYGS\;showed\;38\%$ inhihitory rate on the formation of superoxide at concentration of $0.001\%$ in the human neutrophil. 5. The concentration of inhibiting the production of prostaglandins($PGE_2$) to $11\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS. 6. The concentration of inhibiting the production of interleukins($IL-l{\Beta}$) to $43\%\;and\;39\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS and YSS. 7. HCYGS and YSS didn't influence on collagen synthesis and total protein in fibroblasts. 8. HCYGS and YSS inhibited the collagenase activity to $22\%\;and\;19\%\;at\;0.1\%,\;45\%\;and\;56\%\;at\;0.2\%,\;57\%\;and\;52\%$ at $0.5\%$ respectively, but YSS exerted the inhibitory effect on collagenase activity to $27\%$ at the lower concentration of $0.01\%$. 9. HCYGS and YSS didn't show the faster recovary than control group hut exerted the consistant effect on the anti-inflammations and the formation of granulation tissue.

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