• Title/Summary/Keyword: monooxygenase

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Biotransformation of Aldrin and Chlorpyrifos-methyl by Anabaena sp. PCC 7120

  • Park, Byeoung-Soo;Lee, Sung-Eun
    • Korean Journal of Environmental Agriculture
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    • v.29 no.2
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    • pp.184-188
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    • 2010
  • A cyanobacteria species, Anabaena sp. PCC 7120, was tested to assess its biotransformation ability on two widely used insecticides, aldrin and chlorpyrifos-methyl, in the culture medium. The blue-green alga metabolized aldrin mainly to dieldrin by an epoxidation reaction with the participation of cytochrome P450-dependent monooxygenase in the cyanobacteria. The blue-green alga also produced chlorpyrifosmethyl oxon as a primary metabolite from chlorpyrifos-methyl via a desulfuration reaction, presumably conducted by cytochrome P450-dependent monooxygenase. Therefore, two insecticides might be possibly dissipated by cytochrome P450-dependent monooxygenases in the blue-green algae in the contaminated environments.

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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    • v.24 no.12
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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.

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.

Toluene Monooxygenase의 Peroxide shuntting에 의한 TCE와 PCE 분해 특성

  • 류두현;김형수;최용욱;김용미;이경애;유재수;조현
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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    • pp.277-280
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    • 2004
  • TCE and PCE, suspected carcinogens, are the most common groundwater pollutant from extensive use as a solvent and degreaser. Escherichia coli TGI pBSKAN TOM Green and E. coli TGI pBSKAN ToMO, which were used DNA shuffling technique, produce Toluene-o-monooxygenase(TOM) and toluene-o-xylene- monooxygenase(ToMO). These cells and enzymes are degrading TCE and PCE, TOM and ToMO are needed to cofactor, such as NADH, NADPH and other cofactors. Used TCE and PCE degrading microorganisms experiment the contaminated material removal efficiency. A shunting test used NAD and Hydrogen peroxide.

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Effect of Insecticide Carbofuran and Phenobarbital Sodium and 3-Methylcholanthrene on Activity of Enzyme in Israeli Carp(Cyprinus israeli carpio L.) (살충제 Carbofuran과 Phenobarbital Sodium 및 3-Methylcholanthrene이 이스라엘 잉어의 효소활성에 미치는 영향)

  • Rim, Yo-Sup;Jeong, Jae-Hun;Han, Seong-Soo
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.77-83
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    • 1996
  • Effects of insecticide carbofuran and Phenobarbital sodium(PB) or 3-methylcholanthrene(3-MC) on activities of several enzymes in israeli carps were investigated. Survival number of Israeli carp was the same as that of control when PB and 3-MC only was treated, individually and that was low compared to control when carbofuran only was treated. But survival rate of Israeli carp was high compared to individual treatment of carbofuran when combination treatment of carbofuran and PB or 3-MC was carried out. These results indicate that PB and 3-MC can intervene to detoxify carbofuran exposed to israeli carp. In in vivo test for the effect of this chemicals on activity of enzyme in israeli carp, activities of acetylcholinesterase(AChE) and glutathione S-transferase(GST) were inhibited in carbofuran treatment, but did not in combination treatment of carbofuran and P3 or 3-MC. Activities of UDP-glucuronosyltransfe-rase (UDPGT) and cytochrome P-450-dependent monooxygenase increased in individual or combined treatments of carbofuran and PB or 3-MC. These results suggest that a simultaneous application of carbofuran and PB or 3-MC is critical for the enhancement of activity of AChE, GST, UDPGT and monooxygenase and the protection of Israeli carp from carbofuran toxicity.

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Shuffled toluene-o-xylene monooxygenase를 이용한 TCE 측정용 fluorescence biosensor

  • ;;;;Thomas Wood
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2003.09a
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    • pp.475-478
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    • 2003
  • A sol-gel fiber-optic biosensor with encapsulated pH-sensitive fluorophore and immobilized genetically modified toluene-o-xylene monooxygenase was developed to detect TCE, which is carcinogenic chlorinate organic compounds prevailing in ground water. The sensitivity was characterized for the composition of sol-gel, and manufacturing procedure.

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Characterization of Styrene Catabolic Genes of Pseudomonas putida SN1 and Construction of a Recombinant Escherichia coli Containing Styrene Monooxygenase Gene for the Production of (S)-Styrene Oxide

  • Park Mi-So;Bae Jong-Won;Han Ju-Hee;Lee Eun-Yeol;Lee Sun-Gu;Park Sung-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1032-1040
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    • 2006
  • Some Pseudomonas species can grow on styrene as a sole carbon and energy source. From the new isolate Pseudomonas putida SN1, the genes for styrene catabolism were cloned and sequenced. They were composed of four structural genes for styrene monooxygenase (styA and styB), styrene oxide isomerase (styC), and phenylacetaldehyde dehydrogenase (styD), along with two genes for the regulatory system (styS and styR). All the genes showed high DNA sequence (91% to 99%) and amino acid sequence (94% to 100%) similarities with the corresponding genes of the previously reported styrene-degrading Pseudomonas strains. A recombinant Escherichia coli to contain the styrene monooxygenase from the SN1 was constructed under the control of the T7 promoter for the production of enantiopure (S)-styrene oxide, which is an important chiral building block in organic synthesis. The recombinant E. coli could convert styrene into an enantiopure (S)-styrene oxide (ee >99%) when induced by IPTG The maximum activity was observed as 140 U/g cell, when induced with 1 mM IPTG at $15^{\circ}C$.

Effect of Substrate Size on Activities of Thiocarbamides with the Human Flavin-containing Monooxygenase 3 (사람 Flavin-containing Monooxygenase 3의 Thiocarbamide 화합물의 기질 크기에 따른 효소활성에 관한 연구)

  • 김영미
    • Environmental Analysis Health and Toxicology
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    • v.16 no.2
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    • pp.97-102
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    • 2001
  • The flavin-containing monooxygenases(FMOs) (EC1.14.13.8) are NADPH0dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds, including foods, drugs, pesticides, and other xenobiotics. In humans, FMO3 is quantitatively a major human liver monooxygenase. In the present study, the baculovirus expression vector system was used to overexpress human FMO3 in sect cells for stalytic studies. Microsomes isolated from Spodoptera frugiperda(Sf)9 cells infected with human FMO3 recombinant baculovirus catalyzed the NADPH-and O$_2$-dependent oxidation of methimazole, thiourea, and phenylthiourea. However there was no detectable activity with 1, 3-diphenylthiourea or larger thiocarbamides. Microsomes from control Sf9 cells were devoid of methimazole or thiourea S-oxygenase activity. 1, 3-diphenylthiourea is apparently completely excluded from the catalytic site, these amines drugs are probably approaching the upper size limits of xenobiotics accepted by human FMO3. The substrate specificity of this iosform in humans appears considerably more restriceted than that of pig, guinea pig, rat or rabbit FMO3.

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Effect of activities of monooxygenase, ${\alpha}$, ${\beta}-esterase$ on the degradation of diazinon and dursban in submerged soil (담수토양중 Diazinon과 Dursban의 분해에 미치는 Monooxygenase와 Esterase의 활성)

  • Choi, Jong-Woo;Rhee, Young-Ha;Lee, Kyu-Seung
    • Korean Journal of Environmental Agriculture
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    • v.9 no.2
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    • pp.97-103
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    • 1990
  • In order to determine the major biochemical degradation factors of the two organophosphorus insecticides, diazinon and dursban, the activities of monooxygenase(m. o.) and ${\alpha}$, ${\beta}-esterase$ were studied in submerged soil under laboratory conditions at $30{\pm}1^{\circ}C$ The degradation rate of diazinon by microorganism showed 1.5 times higher than dursban. The m. o. activity increased from 12hrs and 3days after application of diazinon and dursban, respectively. But the ${\beta}-esterase$ activity showed maximum at one day after application of dursban and $5{\sim}8$ days after diazinon application. Also, the ${\beta}-esterase$ activity was about 10 times higher than ${\alpha}-esterase$. Hence, it was concluded that the biological degradation of diazinon was mainly attributed to m. o. activity and the degradation of dursban to ${\beta}-esterase$ activity.

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Biodegradation Study of Gasoline Oxygenates by Butane-Utilizing Microorganisms (부탄 분해 미생물을 이용한 휘발유 첨가제의 분해특성)

  • 장순웅
    • Journal of Soil and Groundwater Environment
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    • v.8 no.1
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    • pp.27-34
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    • 2003
  • In this study, potential degradation of MTBE and other gasoline oxygenates by pure culture ENV425 and mixed culture isolated from gasoline contaminated soil using butane as the sources of carbon and energy was examined and compared. Butane monooxygenases(BMO) of butane-grown ENV425 and mixed culture generated 1-butanol as a major metabolite of butane oxidation and addition of acetylene, specific inhibitor of monooxygenase, inhibited both butane oxidation and 1-butanol production. The results described in this study suggest that alkanes including propane, pentane, and butane are effectively utilized as a growth substrate to oxidize MTBE cometabolically. And also BTEX compounds could be the potential substrate of the MTBE cometabolism. Cell density also affected on the MTBE degradation and transformation capacity(Tc). Increasing cell density caused increasing MTBE degradation but decreased transformation capacity. Other result demonstrated that MTBE and other gasoline oxygenates, ETBE and TAME, were degraded by butane-grown microorganism.