• 제목/요약/키워드: Redox conditions

검색결과 198건 처리시간 0.026초

Modification of N-Terminal Amino Acids of Fungal Benzoate Hydroxylase (CYP53A15) for the Production of p-Hydroxybenzoate and Optimization of Bioproduction Conditions in Escherichia coli

  • Tamaki, Shun;Yagi, Mitsuhiko;Nishihata, Yuki;Yamaji, Hideki;Shigeri, Yasushi;Uno, Tomohide;Imaishi, Hiromasa
    • Journal of Microbiology and Biotechnology
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    • 제28권3호
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    • pp.439-447
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    • 2018
  • The aromatic compound p-hydroxybenzoate (PHBA) is an important material with multiple applications, including as a building block of liquid crystal polymers in chemical industries. The cytochrome P450 (CYP) enzymes are beneficial monooxygenases for the synthesis of chemicals, and CYP53A15 from fungus Cochliobolus lunatus is capable of executing the hydroxylation from benzoate to PHBA. Here, we constructed a system for the bioconversion of benzoate to PHBA in Escherichia coli cells coexpressing CYP53A15 and human NADPH-P450 oxidoreductase (CPR) genes as a redox partner. For suitable coexpression of CYP53A15 and CPR, we originally constructed five plasmids in which we replaced the N-terminal transmembrane region of CYP53A15 with a portion of the N-terminus of various mammalian P450s. PHBA productivity was the greatest when CYP53A15 expression was induced at $20^{\circ}C$ in $2{\times}YT$ medium in host E. coli strain ${\Delta}gcvR$ transformed with an N-terminal transmembrane region of rabbit CYP2C3. By optimizing each reaction condition (reaction temperature, substrate concentration, reaction time, and E. coli cell concentration), we achieved 90% whole-cell conversion of benzoate. Our data demonstrate that the described novel E. coli bioconversion system is a more efficient tool for PHBA production from benzoate than the previously described yeast system.

아연볼의 산화·환원 반응을 통한 연속식 질산성질소 처리에 관한 연구 (A Study on the Removal of Nitrate Nitrogens by Redox Reaction of Zinc Ball)

  • 김준환;김종화;송주영
    • 한국응용과학기술학회지
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    • 제34권3호
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    • pp.487-494
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    • 2017
  • 질산성 질소는 수용액에서 상당히 안정하게 존재하기 때문에 제거하는 데에 있어 상당한 기술이 요구된다. 또한 저 농도의 질산성 질소는 쉽게 제거되는 반면 고농도 질산성 질소는 제거하기가 힘들다. 따라서 본 연구에서는 직경 3mm 정도의 아연 ball을 이용하여 고농도의 질산성 질소를 기체 질소의 형태로 제거하는 것을 목적으로 하였고, 다양한 반응조건에서 질산성 질소의 제거 특성을 실험하였다. 본 연구의 결과로 아연 ball을 사용하여 연속식 처리를 통한 질산성 질소의 처리효율은 약 80%정도 되었다. 하지만 폐수를 수소이온농도 2 정도의 산성 분위기로 유지시켜야하고 처리된 폐수를 중화 처리하여 방류시켜야하는 등의 문제점은 상존하고 있다.

Current- voltage (I-V) Characteristics of the Molecular Electronic Devices using Various Organic Molecules

  • Koo, Ja-Ryong;Pyo, Sang-Woo;Kim, Jun-Ho;Kim, Jung-Soo;Gong, Doo-Won;Kim, Young-Kwan
    • Transactions on Electrical and Electronic Materials
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    • 제6권4호
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    • pp.154-158
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    • 2005
  • Organic molecules have many properties that make them attractive for electronic applications. We have been examining the progress of memory cell by using molecular-scale switch to give an example of the application using both nano scale components and Si-technology. In this study, molecular electronic devices were fabricated with amino style derivatives as redox-active component. This molecule is amphiphilic to allow monolayer formation by the Langmuir-Blodgett (LB) method and then this LB monolayer is inserted between two metal electrodes. According to the current-voltage (I-V) characteristics, it was found that the devices show remarkable hysteresis behavior and can be used as memory devices at ambient conditions, when aluminum oxide layer was existed on bottom electrode. The diode-like characteristics were measured only, when Pt layer was existed as bottom electrode. It was also found that this metal layer interacts with organic molecules and acts as a protecting layer, when thin Ti layer was inserted between the organic molecular layer and Al top electrode. These electrical properties of the devices may be applicable to active components for the memory and/or logic gates in the future.

Epigenetic and Glucocorticoid Receptor-Mediated Regulation of Glutathione Peroxidase 3 in Lung Cancer Cells

  • An, Byung Chull;Jung, Nak-Kyun;Park, Chun Young;Oh, In-Jae;Choi, Yoo-Duk;Park, Jae-Il;Lee, Seung-won
    • Molecules and Cells
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    • 제39권8호
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    • pp.631-638
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    • 2016
  • Glutathione peroxidase 3 (GPx3), an antioxidant enzyme, acts as a modulator of redox signaling, has immunomodulatory function, and catalyzes the detoxification of reactive oxygen species (ROS). GPx3 has been identified as a tumor suppressor in many cancers. Although hyper-methylation of the GPx3 promoter has been shown to down-regulate its expression, other mechanisms by which GPx3 expression is regulated have not been reported. The aim of this study was to further elucidate the mechanisms of GPx3 regulation. GPx3 gene analysis predicted the presence of ten glucocorticoid response elements (GREs) on the GPx3 gene. This result prompted us to investigate whether GPx3 expression is regulated by the glucocorticoid receptor (GR), which is implicated in tumor response to chemotherapy. The corticosteroid dexamethasone (Dex) was used to examine the possible relationship between GR and GPx3 expression. Dex significantly induced GPx3 expression in H1299, H1650, and H1975 cell lines, which exhibit low levels of GPx3 expression under normal conditions. The results of EMSA and ChIP-PCR suggest that GR binds directly to GRE 6 and 7, both of which are located near the GPx3 promoter. Assessment of GPx3 transcription efficiency using a luciferase reporter system showed that blocking formation of the GR-GRE complexes reduced luciferase activity by 7-8-fold. Suppression of GR expression by siRNA transfection also induced down-regulation of GPx3. These data indicate that GPx3 expression can be regulated independently via epigenetic or GR-mediated mechanisms in lung cancer cells, and suggest that GPx3 could potentiate glucocorticoid (GC)-mediated anti-infla-mmatory signaling in lung cancer cells.

Microcrystalline cellulose에 자유 라디칼을 이용한 methyl methacrylate의 그래프팅 반응과 이를 이용한 천연복합필름의 제조 (Grafting of MMA onto MCC through free radical method and its application to all natural cellulose composite film preparation)

  • 이수;박상희;진석환;이선영
    • 한국응용과학기술학회지
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    • 제25권4호
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    • pp.459-468
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    • 2008
  • Methyl methacrylate(MMA) was grafted onto microcrystalline cellulose(MCC) with ceric ammonium nitrate(CAN) as a redox initiator at the various conditions. The cellulose triacetate(CTA) composite films added MCC and MMA-grafted MCC powders were prepared on a glass plate. The graft yield(GY) and graft efficiency(GE) of the grafted MCC were calculated with the simple equations by the weight balance method. The double bond of C=O on the grafted MCC surfaces was confirmed by the fourier transform infrared spectroscopy with attenuated total reflection(FT-IT ATR) spectrophotometer. After grafting, the degree of crystallinity of cellulose powders was decresed by judging from x-ray diffraction(XRD) data. Scanning electron microscope(SEM) photos showed the only solvent and CAN solution could change the roughness of MCC powders and the effect of powder dispersions in composite matrix. The tensile strength of MCC/CTA composite films was decreased with increase of MCC powder contents. When 5% grafted MCC was added, the tensile strength of grafted MCC/CTA composite films was increased from 82.3 MPa to 97.2 MPa. The thermal property of powders was also analyzed by the thermogravimetric analysis(TGA).

생지화학적 환원조건에서 우라늄의 침철석 및 몬모릴로나이트에 대한 수착 특성 (Sorption Characteristics of Uranium on Goethite and Montmorillonite under Biogeochemical Reducing Conditions)

  • 이승엽;조혜륜;백민훈;정의창;정종태
    • 한국광물학회지
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    • 제25권4호
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    • pp.263-270
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    • 2012
  • 산화/환원 반응에 매우 민감한 우라늄의 침철석 및 몬모릴로나이트에 대한 수착 특성을 알아보기 위해 산화우라늄(VI)과 환원우라늄(IV)를 준비하였다. 환원우라늄은 황산염환원박테리아에 의해 황산염이 환원되는 과정에서 같이 환원된 우라늄(IV)를 희석하여 사용하였다. 광물에 대한 우라늄의 수착량은 우라늄(IV)가 우라늄(VI)에 비해 상대적으로 낮았으며, 이러한 원인 중의 하나는 용액상의 우라늄(IV)가 미세한 콜로이드 형태로 존재하여 광물 표면에 대한 수착력이 약했기 때문이다. 투과전자현미경을 사용하여 우라늄(IV)가 나노 콜로이드의 특징을 가지고 있음을 확인하였고, 이러한 결과는 심부 자연계의 지하수를 따라 이동 가능한 우라늄종이 이온성 우라늄(VI)뿐만 아니라 콜로이드성 우라늄(IV)도 포함될 수 있음을 의미한다.

Molecular Characterization of FprB (Ferredoxin-$NADP^+$ Reductase) in Pseudomonas putida KT2440

  • Lee, Yun-Ho;Yeom, Jin-Ki;Kang, Yoon-Suk;Kim, Ju-Hyun;Sung, Jung-Suk;Jeon, Che-Ok;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • 제17권9호
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    • pp.1504-1512
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    • 2007
  • The fpr gene, which encodes a ferredoxin-$NADP^+$ reductase, is known to participate in the reversible redox reactions between $NADP^+$/NADPH and electron carriers, such as ferredoxin or flavodoxin. The role of Fpr and its regulatory protein, FinR, in Pseudomonas putida KT2440 on the oxidative and osmotic stress responses has already been characterized [Lee at al. (2006). Biochem. Biophys. Res. Commun. 339, 1246-1254]. In the genome of P. putida KT2440, another Fpr homolog (FprB) has a 35.3% amino acid identity with Fpr. The fprB gene was cloned and expressed in Escherichia coli. The diaphorase activity assay was conducted using purified FprB to identify the function of FprB. In contrast to the fpr gene, the induction of fprB was not affected by oxidative stress agents, such as paraquat, menadione, $H_2O_2$, and t-butyl hydroperoxide. However, a higher level of fprB induction was observed under osmotic stress. Targeted disruption of fprB by homologous recombination resulted in a growth defect under high osmotic conditions. Recovery of oxidatively damaged aconitase activity was faster for the fprB mutant than for the fpr mutant, yet still slower than that for the wild type. Therefore, these data suggest that the catalytic function of FprB may have evolved to augment the function of Fpr in P. putida KT2440.

Effect of Electrochemical Redox Reaction on Biochemical Ammonium Oxidation and Chemical Nitrite Oxidation

  • Jeon, Bo-Young;Seo, Ha-Na;Kang, Seung-Won;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제20권3호
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    • pp.485-493
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    • 2010
  • A modified graphite felt electrode with neutral red (NR-electrode) was shown to catalyze the chemical oxidation of nitrite to nitrate under aerobic conditions. The electrochemically oxidized NR-electrode (EO-NR-electrode) and reduced NR-electrode (ER-NR-electrode) catalyzed the oxidation of $1,094{\pm}39$ mg/l and $382{\pm}45$ mg/l of nitrite, respectively, for 24 h. The electrically uncharged NR-electrode (EU-NR-electrode) catalyzed the oxidation of $345{\pm}47$ mg/l of nitrite for 24 h. The aerobic bacterial community immobilized in the EO-NR-electrode did not oxidize ammonium to nitrite; however, the aerobic bacterial community immobilized in the ER-NR-electrode bioelectrochemically oxidized $1,412{\pm}39$ mg/l of ammonium for 48 h. Meanwhile, the aerobic bacterial community immobilized on the EU-NR-electrode biochemically oxidized $449{\pm}22$ mg/l of ammonium for 48 h. In the continuous culture system, the aerobic bacterial community immobilized on the ER-NR-electrode bioelectrochemically oxidized a minimal $1,337{\pm}38$ mg/l to a maximal $1,480{\pm}38$ mg/l of ammonium to nitrate, and the community immobilized on the EU-NR-electrode biochemically oxidized a minimal $327{\pm}23$ mg/l to a maximal $412{\pm}26$ mg/l of ammonium to nitrate every two days. The bacterial communities cultivated in the ER-NR-electrode and EU-NR-electrode in the continuous culture system were analyzed by TGGE on the $20^{th}$ and $50^{th}$ days of incubation. Some ammonium-oxidizing bacteria were enriched on the ER-NR-electrode, but not on the EU-NR-electrode.

Purification and Characterization of Laccase from Wood-Degrading Fungus Trichophyton rubrum LKY-7

  • Hyunchae Jung;Park, Chongyawl;Feng Xu;Kaichang Li
    • 한국펄프종이공학회:학술대회논문집
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    • 한국펄프종이공학회 2001년도 춘계학술발표논문집
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    • pp.18-25
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    • 2001
  • A new wood-degrading fungus Trichophyton rubrum LKY-7 secretes a high level of laccase in a glucose-peptone liquid medium. The production of laccase by the fungus was barely induced by 2,5-xylidine. The laccase has been purified to homogeneity through three chromatography steps in an overall yield of 40%. The molecular mass of the purified laccase was about 65 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified laccase had the distinct blue color and had basic spectroscopic features of a typical blue laccase: two absorption maxima at 278 and 610 nm and a shoulder at 338 nm. The N-terminus of the laccase has been sequenced, revealing high homology to laccases from wood-degrading white-rot fungi such as Ceriporiopsis subvermispora. The enzyme had a "low" redox potential (0.5 V vs normal hydrogen electrode), yet it was one of the most active laccases in oxidizing a series of representative substrates/mediators. Compared with other fungal laccases, the laccase has a very low Km value with ABTS [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid] as a substrate and a very high Km value with violuric acid as a substrate. The laccase has the isoelectric point of 4.0. The laccase had very acidic optimal pH values (pH 3-4) while it was more stable at neutral pH than at acidic pH. The laccase oxidized hydroquinone faster than catechol and pyrogallol. The oxidation of tyrosine by the laccase was not detectable under the reaction conditions. The laccase was strongly inhibited by sodium azide and sodium fluoride. fluoride.

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열화학적 수소 제조 기술 (Themochemical Cycles for Hydrogen Production from Water)

  • 김종원;박주식;황갑진;배기광
    • 에너지공학
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    • 제15권2호
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    • pp.107-117
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    • 2006
  • 물을 분해하여 수소를 만드는 방법으로서 열화학싸이클을 이용한 방법에 대하여 그동안의 연구 동향에 대하여 살펴보았다. 수소생산이란 관점에서 열화학싸이클이 갖는 장점은 일정한 고온의 열을 얻을 수 있다면, 반응속도의 향상과 아울러 대용량화가 가능하다는 점이다. 안정한 물을 분해하려면 물의 산화/환원이 용이한 매개체를 써서 수소 및 산소를 발생하게 하고 순환시키게 되는데, 매개체가 유독성 물질이라면 이 과정에서 누출이 되지 않도록 하여야 한다. 아직 상용화단계에는 미치지 못하였지만, 일본, 스위스, 이스라엘, 미국, 한국 등에서 집중적으로 연구되고 있는 내용은 IS 싸이클과 ZnO/Zn, $Fe_3O_4/FeO$등과 같은 금속산화물계를 이용한 싸이클들이며, 고온용 및 내부식성 소재와 시스템 분야에서 아직 해결해야할 점이 많다.