• 제목/요약/키워드: OPH

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Characteristics of Parathion Hydrolase by Pseudomonas rhodesiae H5 (Pseudomonas rhodesiae H5가 생산하는 Parathion Hydrolase의 특성)

  • Yun Nam Kyung;Park Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.199-204
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    • 2004
  • The parathion hydrolase (OPH) produced by Pseudomonas rhodesiae H5 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Parathion hydrolase from crude extracts of P. rhodesiae H5 has two components designated as OPH $I_1$ and OPH $I_2$, Optimum pH and temperature of OPH $I_1$and OPH $I_2$ were pH 7.2 and $30^{\circ}C$, and pH 7.6 and $37^{\circ}C$, respectively. The activation energy of OPH $I_1$ for the hydrolysis of parathion was 3.01 ㎉/I, II, III in the temperature range of $4^{\circ}C$ to $30^{\circ}C$, and Michaelis constant ($K_m$) for parathion was 69.2 ${\mu}M$. The activation energy of OPH $I_2$ for the hydrolysis of parathion was 4.07㎉/㏖ in the temperature range of $4^{\circ}C$ to $37^{\circ}C$, and Michaelis constant for parathion was 150.9${\mu}M$. Furthermore OPH $I_1$ was completely inhibited by 1 mM $Ca^2+$, $Cu^2+$, $Mg^2+$, $Ni^2+$, but OPH $I_2$ was less inhibited than OPH $I_1$ by the metals used in this study.

Biodegradation of Organophosphate Pesticide Using Recombinant Cyanobacteria with Surface- and Intracellular-Expressed Organophosphorus Hydrolase

  • Chungjatupornchai, Wipa;Fa-Aroonsawat, Sirirat
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.946-951
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    • 2008
  • The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.

Effect of Addition of Tween 20 and Glycerol in Recombinant Escherichia Coli Culture on Organophosphorus Hydrolase (OPH) Production for Biodrgradation of Coumaphos Insecticide (Coumaphos 살충제의 생분해를 위하여 재조합 대장균 배양에서 Tween 20과 Glycerol 첨가가 유기인분해 효소 생산에 미치는 영향)

  • Choi, Suk Soon;Seo, Sang Hwan
    • Applied Chemistry for Engineering
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    • v.18 no.5
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    • pp.501-505
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    • 2007
  • Organophosphorus hydrolase (OPH) expressed from recombinant Escherichia coli was used to biodegrade organophosphate insecticide coumaphos which has a very high toxicity in mammalian cells. To improve the productivity of OPH, the effects of nonionic surfactants (Tween 20, PEG 1000) and organic solvents, such as glycerol, propanol, and ethanol, were investigated in the strain culture. The maximum OPH was produced when the 0.25% of Tween 20 and 0.5% of glycerol were added to the medium. As the OPH obtained from disrupt-cell process by ultrasound treatment was used, the biodegradation efficiencies of 0.2, 0.5, 1.0 and 2.0 mM coumaphos were 100, 88, 84 and 78%, respectively. A novel method developed in this study could be applied to the biodetoxification technology in the contaminated region with various coumaphos concentration.

Enhancement of Paraoxon Biodegradation Rate from Recombinant Escherichia coli Catalyst for Bioremediation (Bioremediation을 위하여 재조합 대장균 촉매를 이용한 Paraoxon의 생분해 속도 향상)

  • Choi, Suk Soon;Seo, Sang Hwan;Kang, Dong Gyun;Cha, Hyung Joon;Yeom, Sung Ho
    • Journal of the Korea Organic Resources Recycling Association
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    • v.14 no.3
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    • pp.110-116
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    • 2006
  • In this study, the biodegradation rate of paraoxon, that is an organophosphate pesticide, was enhanced by recombinant Escherichia coli harboring organophosphorus hydrolase (OPH). The optimum conditions were 8.5 of initial pH and 5.0% of acetone for the enhancement of specific whole cell OPH activity. When the OPH was produced to 498 Unit/L, 98% of 275mg/L paraoxon was degraded within 10 minutes, and thus the biodegradation rate was enhanced to $29.2mg/g{\cdot}min$. The results implied that practical bioremediation technology developed in this study was an effective method to degrade residual organophosphate pesticide in ground water or soils in a short time.

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Production of Periplasmic Space-Secreted Organophosphorus Hydrolase from Recombinant Escherichia coli for Degradation of Environmental Toxic Organophosphate Compounds (환경 독성 유기인 화합물 분해를 위하여 재조합 대장균에서 세포내 간극으로 분비된 Organophosphorus Hydrolase의 생산)

  • Choi, Suk Soon;Seo, Sang Hwan;Kang, Dong Gyun;Cha, Hyung Joon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.3
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    • pp.89-96
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    • 2005
  • In the present work, production of organophosphorus hydrolase (OPH) that is secreted in periplasmic space of recombinant Escherichia coli was performed for degradation of environmental toxic organophosphate compounds, paraoxon. The optimal conditions for enhancement of OPH production were 1.0 mM isopropyl-${\beta}$-D-thiogalactopytanoside (IPTG), 0.25 mM $Co^{2+}$, and 0.1 mM ethylenediamine tetraacetate (EDTA). Under these culture conditions, the maximum OPH production was $174Unit/L{\cdot}OD$. In addition, 1 mM of paraoxon was completely degraded by OPH. These results can be used as a bioremediation tool for removal of environmental toxic organophosphate compounds remaining in soil and aquatic environment.

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Orange Phosphorescent Organic Light-emitting Diodes Using a Spirobenzofluorene-type Phospine Oxides as Host Materials

  • Jeon, Young-Min;Lee, In-Ho;Lee, Chil-Won;Lee, Jun-Yeob;Gong, Myoung-Seon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.10
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    • pp.2955-2960
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    • 2010
  • Spiro-type orange phosphorescent host materials, 9-diphenylphosphine oxide-spiro[fluorene-7,9'-benzofluorene] (OPH-1P) and 5-diphenylphosphine oxide-spiro[fluorene-7,9'-benzofluorene] (OPH-2P) were successfully prepared by a lithiation reaction followed by a phosphination reaction with diphenylphosphinic chloride. The EL characteristics of OPH-1P and OPH-2P as orange host materials doped with iridium(III) bis(2-phenylquinoline)acetylacetonate ($Ir(pq)_2acac$) were evaluated. The electroluminescence spectra of the ITO (150 nm)/DNTPD (60 nm)/NPB (30 nm)/OPH-1P or OPH-2P: $Ir(pq)_2acac$ (30 nm)/BCP (5 nm)/$Alq_3$ (20 nm)/LiF (1 nm)/Al (200 nm) devices show a narrow emission band with a full width at half maximum of 75 nm and $\lambda_{max}$ = 596 nm. The device obtained from OPH-1P doped with 3% $Ir(pq)_2acac$ showed an orange color purity of (0.580, 0.385) and an efficiency of (14 cd/A at 7.0 V). The ability of the OPH-P series to combine a high triple energy with a low operating voltage is attributed to the inductive effect of the P=O moieties and subsequent energy lowering of the LUMO, resulting in the enhancement of both the electron injection and transport in the device. The overall result is a device with an EQE > 8% at high brightness, but operating voltage of less than 6.4 V, as compared to the literature voltages of ~10 V.

Housing Lifestyle of One Person Households with the Perspective of Digital Placemaking (디지털화에 따른 1인 가구의 주생활양식에 관한 연구)

  • Cho, Sung-Heui;Park, Soo-Been;Hwang, Mi-Ri
    • Proceeding of Spring/Autumn Annual Conference of KHA
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    • 2003.11a
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    • pp.137-143
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    • 2003
  • This study aimed to analyze the housing lifestyles of one person households (OPH) with the perspective of digital placemaking in order to suggest the planning criteria for OPH house. The structured questionnaire survey was accomplished by 145 OPH. The main findings were as follows. (1) OPH's housing lifestyles were classified into four types composed of ten factors. (2) The user preference for the space organization was varied by four types of housing lifestyles. (3) Most OPH required the informational appliances and their needs for other digital appliances were significantly differed by four types of housing lifestyles.

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PERIOD VARIATION STUDY OF THE A-TYPE W UMA ECLIPSING BINARY V839 OPH

  • Hanna, Magdy A.
    • Journal of The Korean Astronomical Society
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    • v.43 no.6
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    • pp.201-211
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    • 2010
  • We present an analysis of the measurements of mid-eclipse times of V839 Oph, collected from literature sources. Our analysis indicates a period increase of $3.2{\times}10^{-7}$ day/yr. This period increase of V839 Oph can be interpreted in terms of mass transfer of rate $1.76{\times}10^{-7}M_{\odot}/yr$, from the less to the more massive component. The O - C diagram shows a damping sine wave covering two different complete cycles of 36.73 yr and 19.93 yr with amplitudes approximately equal to 0.0080 and 0.0043 day, respectively. The third cycle has to be expected to cover about 13.5 years with lower amplitude than those of the former two cycles. These unequal duration cycles show a non periodicity which may be explained as resulting from either the presence of a tertiary component to the system or cyclic magnetic activity variations due to star spots. For the later mechanism, the obtained characteristics are consistent when applying Applegate (1992) mechanism.

Surface Display of Organophosphorus Hydrolase on E. coli Using N-Terminal Domain of Ice Nucleation Protein InaV

  • Khodi, Samaneh;Latifi, Ali Mohammad;Saadati, Mojtaba;Mirzaei, Morteza;Aghamollaei, Hossein
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.234-238
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    • 2012
  • Recombinant Escherichia coli displaying organophosphorus hydrolase (OPH) was used to overcome the diffusion barrier limitation of organophosphorus pesticides. A new anchor system derived from the N-terminal domain of ice-nucleation protein from Pseudomonas syringae InaV (InaV-N) was used to display OPH onto the surface. The designed sequence was cloned in the vector pET-28a(+) and then was expressed in E. coli. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by InaV-N on the outer membrane, and the ability of recombinant E. coli to utilize diazinon as the sole source of energy, without growth inhibition, indicated its significant activity. The location of OPH was detected by comparing the activity of the outer membrane fraction with the inner membrane and cytoplasm fractions. Studies revealed that recombinant E. coli can degrade 50% of 2 mM chlorpyrifos in 2 min. It can be concluded that InaV-N can be used efficiently to display foreign functional protein, and these results highlight the high potential of an engineered bacterium to be used in bioremediation of pesticide-contaminated sources in the environment.