• Title/Summary/Keyword: E. coli concentration

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Purification and Characterization of Bacillus subtilis Protoporphyrinogen Oxidase and Pre-equilibrium Behavior During Oxidation of Protoporphyrinogen IX

  • Jeong, Eun-Ju;Han, Ok-Soo
    • BMB Reports
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    • v.34 no.1
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    • pp.39-42
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    • 2001
  • Previous studies indicate that B. subtilis protoporphyrinogen oxidase is poorly inhibited by diphenyl ether herbicides. To better understand the basis of this insensitivity, the enzyme was overexpressed as a soluble protein in E. coli, purified and characterized. The mechanism of oxidation of B. subtilis protoporphyrinogen IX was studied and the enzyme kinetic parameters were determined for protoporpyrinogen IX; $K_m$, and $k_{cat}$ were $6.3\;{\mu}M$ and $0.028\;h-^1$, respectively. The enzyme required flavin adenine dinucleotide as a cofactor and its activity was enhanced by 1 mM n-octylglucopyranoside. The nonenzymatic oxidation rate was dependent on the concentration of protoporphyrinogen IX, suggesting that the reaction involves a pre-equilibrium step followed by a rate-limiting step.

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Characteristics of Trichloroethene and Tetrachloroethene Sensing Optical Fiber Biosensor Using Toluene-o-monooxygenase and Fluoresceinamine (Toluene-o-monooxygenase와 Fluoresceinamine을 이용한 Trichloroethene와 Tetrachloroethene 감지용 광섬유 바이오센서의 특성)

  • Ryoo, Doohyun
    • Journal of Soil and Groundwater Environment
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    • v.23 no.4
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    • pp.42-47
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    • 2018
  • E. coli TG1 pBS TOM Green was cultured to produce toluene-o-monooxygenase (TOM). A biosensor system was successfully constructed using purified TOM to effectively detect trichloroethene (TCE) and tetrachloroethene (PCE), which represent some of the major contaminants in groundwater and soil. In order to utilize TOM as a sensor, NADH, a biological oxidizer, was replaced with hydrogen peroxide which is a chemical oxidizing agent. A three-layered sandwich-type sensing tip was fabricated on the outside of the hydrophilic polyvinylidene fluoride membrane. TCE and PCE were applied to the sensor and the hydrogen ions were measured by a fiber optic fluorometer using fluoresceinamine. Calibration curves were obtained for TCE and PCE in the concentration range of 0.2-100 mg/l, and the detection limit of the system was $10{\mu}g/l$ for TCE and PCE.

Antimicrobial activity of sophorolipid

  • Kim, Gab-Jung;Kim, Young-Bum;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.247-249
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    • 2000
  • Antimicrobial activity of sophorolipid produced by Candida bombicola ATCC 22214 was investigated, Minimum inhibitory concentration of sophorolipid to Propionibacterium acne and Bacillus subtilis were 0.5 ppm and 4 ppm, respectively. However no antimicrobial activity to E. coli was detected. Indicating the antimicrobial activity of sophorolipid only toward gram positive bacteria. Antifungal activity of sophorolipid was also detected in the hyphae growth assay for Botrytis cinerea. Malate dehydrogenase, a cytoplasmic enzyme, of B. subtilis was remarkably increased with the sophorolipid treatment, indicating the leakage of membrane by sophorolipid. This result shows the potentials of sophorolipid as mild, non-toxic antimicrobial agent.

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Effects and Optimization of Gamma-Amino Butyric Acid (GABA) Production Process using Glutamate Decarboxylase (GAD) (Glutamate Decarboxylase (GAD)를 이용한 Gamma-Amino Butyric Acid (GABA) 생산 및 최적화)

  • Kim, Eui Jin;Lee, Jung-Heon
    • KSBB Journal
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    • v.29 no.6
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    • pp.426-431
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    • 2014
  • pH controlled batch reactor and bubble column reactors have been developed in this research. They were used to produce high concentration of GABA and to determine optimal pH for GABA production. Glutamate decarboxylase (GAD) was isolated from recombinant E. coli and used for GABA production from monosodium glutamate (MSG). pH control was inevitable because the pH increased with MSG consumption. GAD showed highest activity at acidic conditions at pH 5.5 but the optimal pH for GABA production was pH 6.0. When 1.5 mole of MSG was used as reactant, the 1.05 mole of GABA was produced after 10 hrs batch reaction. Using bubble column reactors, 80 % of MSG was converted to GABA for 6 hrs reaction and 1.2 mole of GABA was produced.

On-line monitoring of microorganism cultivation processes using optical sensing membranes for simultaneous detection of dissolved oxygen and pH (용존산소와 pH의 동시 검출용 광학 센서 막을 이용한 미생물 발효공정의 온라인 모니터링)

  • Kim, Chun-Kwang;Rhee, Jong-II
    • KSBB Journal
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    • v.24 no.1
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    • pp.106-112
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    • 2009
  • An optical sensing membrane has been fabricated to measure the concentration of dissolved oxygen(DO) and pH value simultaneously. It has employed HPTS as a pH sensitive dye and a ruthenium(II) complex as a DO sensitive dye. The sensing membrane has been applied to wells in a 24-well microtiter plate. Using the 24-well microtiter plate the concentrations of dissolved oxygen and pH values have been on-line monitored during the cultivations of E.coli DH5${\alpha}$, B.cereus 318 and P.pastoris X-33. On-line monitoring of DO and pH in microorganism cultivation processes showed good performance of the sensing membrane containing 5 mM HPTS and 2 or 5 mg/mL Rudpp.

Antimutagenic Effects of Linoleic Acid

  • Lim, Sun-Young;Rhee, Sook-Hee;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.2 no.1
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    • pp.29-34
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    • 1997
  • In order to determine the effectiveness of linoleic acid(LA) to inhibit carcinogens/mutagens-induced mu-tagenesis, Ames test using Salmonella typhimurium TA100 and the SOS chromotest using E. Coli PQ37, were carried out. The inhibitory effect of LA(1%) on the Ames mutagenicity test were 98%, 78%and 69% mediated by aflatoxin B₁(AFB₁), N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitroquinoline-1-oxide(4-NQO), respectively. LA exhibited a strong antimutagenic activity aganist indirect mutagen, AFB₁whereas exhibited the same concentration of LA showed weaker inhibitory effects on direct mutagens of MNNG and 4-NQO than that AFB₁. LA also reduced the SOS responses induced by MNNG and 4-NQO significantly. This result showed a possibility that LA can be a protective agent in early step of cancinogenesis.

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Free Radical Involvement in the DNA Damaging Activity of Fumonisin Bl

  • Lee, Wan-Hee;Lee, Kil-Soo
    • Toxicological Research
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    • v.17 no.4
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    • pp.249-253
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    • 2001
  • Fumonisin B1, a mycotoxin, is thought to induce esophageal cancer in humans and apoptosis in animal cells by inhibiting ceramide synthase. Dumonisin Bl may also generate reactive oxygen species directly or indirectly, leading to DNA damage and lipid peroxidation. In this study, a DNA fragmentation assay, dichlorofluorescein (DCF) analysis, and single cell gel electrophoresis (SCGE) were used to investigate the involvement of cellular free radicals, specifically hydrogen peroxide, in the DNA damaging activity of fumonisin B1. From an in vitro DNA fragmentation assay, E. coli DNA, damage by fumonisin Bl was increased by the addition of superxide dismutase (SOD) and decreased by catalase. SCGE and DCF analysis in vivo showed that the nuclear DNA damage and intracellular free radicals in cultured rat hepatocytes treated with fumonisin B1 were increased with the concentration of fumonisin Bl . DNA damage and free radical generation were inhibited by the addition of catalase. Fumonisin Bl , in the presence of SOD, produces hydrogen peroxide causing oxidative DNA damage and protein malfunction, leading to genotoxicity and cytotoxicity of the toxin.

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Rifampicin에 내성인 Bifidobacterium bifidum 균주 개발

  • 최웅칠;고성열;김병각
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.143-143
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    • 1993
  • 현재 시판되고 있는 정장용 생균 제제에 함유되어있는 정장 균주의 하나인 Bifidobacterium bifidum은 항결핵제 중 rifampicin에 감수성으로 rifampicin과 병용 투여시 본래의 정장 효과를 기대할 수없다. 따라서, rifampicin에 내성인 돌연변이 균주를 얻기 위해 B. bifidum을 N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)로 처리하여 rifampicin에 내성인 30 종의 균주를 선별하였고, rifampicin에 대한 Minimal Inhibitory Concentration (MIC)를 측정해 본 결과 내성이 1,000 배 이상 상승하였다. 균주 동정을 위하여 fructose-6-phosphate phosphoketolase test를 실시해 본 결과 Bifidobacterium임이 확인되었다. 이들 내성 균주들의 유기산 생산량을 측정하여 그 생산량이 모균주와 가장 유사한 3 종의 균주를 선발하였다. 이들에 대하여 Escherichia coli 생육 억제능을 시험해 본 결과 E. coil 생육 억제능이 모균주와 유사하였다. 또, rifampicin을 함유한 배지에서 돌연변이 균주를 배양시킨 경우 rifampicin이 안정한 상태로 잔존한 것을 알 수 있었다. 이것으로 보아 돌연변이 균주들은 rifampicin을 분해 또는 변형시키는 효소를 생산하지 않는다고 볼 수 있다. 이상의 결과로 본 연구에서 개발한 돌연변이 균주들, 즉 B. bifidum RFRll, RFR21 그리고 RFR61은 rifampicin에 내성이면서 모균주와 동일한 생화학적 특성을 갖는 정장 균주로 여겨진다.

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Monitoring of Indicator Microorganism Concentrations of River Sediment and Surface Water in the Geum River Basin (금강 수계 내 하천퇴적물 및 지표수의 지표미생물 농도분포)

  • Kim, Geonha
    • Journal of Korean Society on Water Environment
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    • v.26 no.1
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    • pp.125-132
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    • 2010
  • Characterization of sediment quality is important for the proper management of surface water quality, yet sediment has not been monitored sufficiently. In this study, fecal indicator microorganism concentrations of sediments in the Geum River Basin were monitored. Sampling was carried out at one paddy field, one lakeshore and five monitoring stations in the lower reach of the Geum River Basin. Surface waters and sediments were sampled four times during rainy season. Total coliform concentrations of sediments were 12 times higher in average to those of surface waters while E. coli concentrations of sediments were six times higher. No correlation found between indicator microorganism concentration between surface waters and sediments.

Allelopathic Potential and Substances from Cork Tree (Pbellodendron amurense Rupr.)

  • Park, Young-Goo;Choi, Myung-Suk;Yang, Jae-Kyung;Paik, Ki-Hyon
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.3
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    • pp.92-98
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    • 2001
  • Allelopathic effects of the cork tree (Phellodendron amurense Rupr.) on several crops and soil miro-organisms were assessed using germination bioassay and antimicrobial assay, and allelochemicals were identified. In a germination bioassay, extract of cork tree inhibited at high concentration on germination of several crop seeds such as cabbage, lettuce, and cucumber. However, aqueous extracts inhibited powerfully growth of test organisms such as Streptococus aureus, S. aureus, S. typhimurium, and E. coli as bacteria, and Candida albicans as yeast, and Botrytis cineria and Alternata alternaria as fungi.. The cork tree extract showed strong antimicrobial activities against isolated soil fungi. The allelochemicals were separated using Silica gel, Sephadex LH-20 gel column chromatography and HPLC. The substances were analyzed by UV spectrometry and EI-mass spectrometry. The active allelochemicals were identified as isoquinoline alkaloids, berberine and palmatine.

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