• 제목/요약/키워드: organophosphorus compounds

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유기인화합물 측정용 광바이오센서 개발 (Development of Prototype Biosensor for The Detection of Organophosporus Compounds)

  • 최정우;김종민;이원홍;김영기
    • KSBB Journal
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    • 제17권2호
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    • pp.158-161
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    • 2002
  • 본 연구에서는 화학무기, 농약 등에 사용되는 신경독성물질인 유기인화합물의 측정을 위하여 유기인화합물의 효소반응 저해작용을 이용한 광바이오센서장치의 시제품을 제작하였다. 효소반응을 위하여 효소로는 신경세포의 필수효소인 acetylcholinesterase, acetylthiocholine iodide을 사용하였으며 효소반응의 저해제인 유기인화합물로는 paraoxon을 사용하였다. 센서의 폭정원리는 유기인화합물에 의해 저해된 효소반응정도를 효소반응의 생성물인 아세트산의 정량적 측정으로 분석하였으며, pH에 의하여 최대 흡광파장의 변화가 일어나는 litmus를 사용하여 흡광도 측정으로 아세트산의 정량분석을 수행하였다. 광바이오센서 시제품의 제작은 광원으로 고취도 LED와 광세기 측정을 위한 photodiode로 구성하였으며, 제작된 센서를 이용한 실험결과로부터 0 ppm에서 2 ppm의 paraoxon 농도에서 구성된 센서시스템의 선형적 신호 변화를 관찰하였다. 이상의 실헐결과로부터 광바이오센서 시제품은 2분의 반응시간으로 신속하고 정확한 유기인화합물의 정량분석이 가능함을 확인하였다.

An Evaluation of Liquid Chromatography/Mass Specrometry with Atmospheric Pressure Chemical Ionizarion for the Rapid and Simultaneous Measurement of Carbamate Pesticides and Organophosphorus Pesticides

  • 김병주;소현영
    • Bulletin of the Korean Chemical Society
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    • 제21권5호
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    • pp.471-476
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    • 2000
  • Liquid chromatography/mass spectrometry with an atmospheric pressure chemical ionization interface (LC/APCI/MS) is evaluated for the simultaneous determination of carbamate pesticides and organophosphorus pesticides in a single chromatographic analysis. APCI mass spectra of those compounds were obtrained to study their ionization characteristics. APCI provided abundant ions such as protonated molecules and characteristic fragment ions for carbamate pesticides and organophosphorus pesticides. To evaluate the feasibility of the LC/APCI/MS for a routine quantitative analysis, the linearity and repeatability of LC/APCI/MS were examined by measuring standard solution mixtures of five carbamate pesticides and four organophosphorus pesticides over the range of 1 to 100 ㎍/mL. Teh peak areas in chromatograms of characteristic ions for those compounds showed less than 3% of variation from run to run. The standard calibration curves for the nine pesticides show good linearity in the concentration range. The detection limits of the LC/APCI/MS system for those compounds range from 0.006 to 0.2 ng.

유기인화합물 분해효소 유전자의 재조합 및 단백질 발현 (Subcloning and Expression of a Gene Encoding an Organophosphorus Acid Anhydrolase)

  • 박재왕;김석찬;이남택
    • 한국군사과학기술학회지
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    • 제4권1호
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    • pp.188-197
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    • 2001
  • Organophosphorus acid anhydrolases(OPAA) catalyzing the hydrolysis of toxic organophosphates have been found in a variety of prokaryotic and eukaryotic organisms. Of the several kinds of OPAA that can degrade nerve agents, such as DFP, sarin and soman, a OPAA gene harbored in the chromosomal DNA of Alteromonas haloplanktis strain was subcloned in order to develope an enzymatic degradation method of toxic organophosphorus compounds. For this 1481 bp DNA fragment containing OPAA gene and its flanking regions has been synthesized through PCR using chromosomal DNA of A. haloplanktis strain. After subcloning and subsequent expression, crude OPA anhydrolase was prepared and assayed. It was shown that the OPAA had a very high hydrolytic activity on DFP. The specific activity of the enzyme was 1,110 $\mu$mole.$min^{p-1}.mg^{-1}$ protein. It seemed that OPAA with such a high hydrolytic activity may give a good prospects to its use, as a biodegradation tool, in detoxifying toxic organophosphorus compounds, such as pesticides and chemical stockpiles which are posing a potential threat to the field environment and human health.

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유기인 계열 독성화합물 분해를 위한 촉매반응의 최신 연구 동향 (Recent Trend in Catalysis for Degradation of Toxic Organophosphorus Compounds)

  • 계영식;정근홍;김동욱
    • 공업화학
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    • 제30권5호
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    • pp.513-522
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    • 2019
  • 유기물질, 전이금속 및 유기-금속 구조체(MOFs)를 기반으로 하는 촉매들이 유기인 계열 독성물질들을 분해하고 제거하는데 효과적임이 보고되어 왔다. 최근 20년간 독성물질 분해연구를 위해 다공성 MOFs들이 응용 목적에 맞게 디자인되고 합성되었다. $Zr_6$ 기반의 금속노드와 유기결합체를 가지는 MOFs들은 세공크기, 공극률, 표면적, Lewis acidic 자리 그리고 열적 안정성 등과 같은 기본구조내의 변형이 가능하기 때문에 화학작용제, 살충제 및 제초제를 제거하는 촉매로 널리 사용되어왔다. 본 리뷰에서는 구조, 안정성, 입자크기, 연결된 리간드 수, 유기 기능기 등에 따른 MOFs들의 촉매효율과의 연관성을 다루게 될 것이다.

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

  • 최석순;서상환;강동균;차형준
    • 유기물자원화
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    • 제13권3호
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    • pp.89-96
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    • 2005
  • 본 연구에서는 유기인 화합물인 Paraoxon의 분해를 위하여 재조합 대장균에서 세포내 간극(periplasmic space)으로 분비되는 organophosphorus hydrolase(OPH)의 생산에 대하여 고찰하였다. OPH 생산의 향상을 위하여 성장 배지에 첨가되는 최적의 조건은 1.0 mM isopropyl-${\beta}$-D-thiogalactopytanoside (IPTG), 0.25 mM $Co^{2+}$ 및 0.1 mM ethylenediamine tetraacetate (EDTA) 이었다. 이 조건에서 최대OPH 생산은 $174Unit/L{\cdot}OD$를 나타내었다. 또한 1 mM의 Paraoxon은 OPH에 의하여 완전히 분해되었다. 이러한 연구 결과는 토양 및 수계에 잔류하는 환경독성 유기인 화합물을 제거하는 bioremediation의 수수단으로 활용될 수 있음을 보여주었다.

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Detoxification of Sarin, an Acetylcholinesterase Inhibitor, by Recombinant Organophosphorus Acid Anhydrolase

  • Kim, Seok-Chan;Lee, Nam-Taek
    • BMB Reports
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    • 제34권5호
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    • pp.440-445
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    • 2001
  • Pesticide waste and chemical stockpiles are posing a potential threat to both Vie environment and human health. There is currently a great effort toward developing effective and economical methods for the detoxification of these toxic organophosphates. In terms of safety and economy, enzymatic biodegradation has been recommended as the most promising tool to detoxify these toxic materials. To develop an enzymatic degradation method to detoxify such toxic organophosphorus compounds, a gene encoding organophosphorus acid anhydrolase (OPAA) from genomic DNA of Alteromonas haloplanktis C was subcloned and expressed. The enzyme consists of a single polypeptide chain with a molecular weight of 48 kDa. It demonstrates strong hydrolyzing activity on sarin, an acetylcholinesterase inhibitor. Moreover, its high activity is sustained for a considerable length of time. It is projected that the recombinant OPAA can be applied as an enzymatic tool that can be used not only for the detoxification of pesticide wastes, but also for the demilitarization of chemical stockpiles.

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유기인계 화합물의 산화대사중 반응성 중간체와 반응기작에 관한 고찰 (Reactive Intermediates and Reaction Mechanisms in the Oxidative Metabolism of Organophosphorus Compounds)

  • 김정한;;박창규
    • 한국환경농학회지
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    • 제15권2호
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    • pp.246-261
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    • 1996
  • 현재 사용되고 있는 유기합성 농약의 중요한 부분을 차지하고 있는 유기인계 농약의 대부분은 분자에 황 원자를 포함하고 있으며 환경이나 생체 내에서 대사되어 활성화 되거나 무독화 된다. 이에 관련된 다양한 반응 중 산화반응이 주요 대사반응으로 여겨지는데 이산화대사에 의해 생성된 대사물의 특성도 중요하거니와 반응 중 생성되는 반응성 중간체에 대한 연구는 대사반응의 경로 및 기작 구명에 필수적이고 농약의 독성학적 특성을 이해하는 데에 중요한 위치를 차지하고 있다. 본 총설은 유기인계 농약을 중심으로 관련 유기인 화합물을 대상으로 산화대사 반응 중 생성되는 반응성 중간체에 대한 연구들을 그 구조구명과 반응 기작면에서 다루었다.

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LC/MS/MS를 이용한 유기인계 농약류의 최적 분석법 정립과 원·정수에서의 모니터링 (Development of Analytical Method and Monitoring of Organophosphorus Pesticides in the Raw Water and Clean Water by Liquid Chromatography-Tandem Mass Spectrometry)

  • 김경아;송미정;염훈식;손희종;이상원;최진택
    • 한국환경과학회지
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    • 제24권12호
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    • pp.1569-1582
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    • 2015
  • The analytical method for 16 organophosphorus pesticides was developed in this study. The 16 organophosphorus pesticides were analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) using on-line solid phase extraction (on-line SPE) with PLRP- S cartridge. Analysis of all analytes in the MS/MS was processed in the electrospray ioni-zation (ESI) positive mode. They are Azinphos ethyl, Chlorfenvinphos, Ethion, Famphur, Phosmet, Phosphamidon, Terbufos, Aspon, Chlorpyrifos-methyl, Crotoxyphos, Dichlofenthi-on, Dicrotophos, Fonofos, Thionazin, Dimethoate and Iprobenfos. Limits of detection (LODs) and Limits of quantification(LOQs) were obtained as 0.8~2.0 ng/L and 2.6~6.4 ng/L, respectively. All compounds were not detected at the 8 sampling points of the raw water and clean water.

Engineered Recombinant PON1-OPH Fusion Hybrids: Potentially Effective Catalytic Bioscavengers against Organophosphorus Nerve Agent Analogs

  • Lee, Nari;Yun, Hyeongseok;Lee, Chan;Lee, Yikjae;Kim, Euna;Kim, Sumi;Jeon, Hyoeun;Yu, Chiho;Rho, Jaerang
    • Journal of Microbiology and Biotechnology
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    • 제31권1호
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    • pp.144-153
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    • 2021
  • Organophosphorus nerve agents (OPNAs), including both G- and V-type nerve agents such as sarin, soman, tabun and VX, are extremely neurotoxic organophosphorus compounds. Catalytic bioscavengers capable of hydrolyzing OPNAs are under development because of the low protective effects and adverse side effects of chemical antidotes to OPNA poisoning. However, these bioscavengers have certain limitations for practical application, including low catalytic activity and narrow specificity. In this study, we generated a fusion-hybrid form of engineered recombinant human paraoxonase 1 (rePON1) and bacterial organophosphorus hydrolase (OPH), referred to as GV-hybrids, using a flexible linker to develop more promising catalytic bioscavengers against a broad range of OPNAs. These GV-hybrids were able to synergistically hydrolyze both G-type OPNA analogs (paraoxon: 1.7 ~ 193.7-fold, p-nitrophenyl diphenyl phosphate (PNPDPP): 2.3 ~ 33.0-fold and diisopropyl fluorophosphates (DFP): 1.4 ~ 22.8-fold) and V-type OPNA analogs (demeton-S-methyl (DSM): 1.9 ~ 34.6-fold and malathion: 1.1 ~ 4.2-fold above) better than their individual enzyme forms. Among the GV-hybrid clones, the GV7 clone showed remarkable improvements in the catalytic activity toward both G-type OPNA analogs (kcat/Km (106 M-1 min-1): 59.8 ± 0.06 (paraoxon), 5.2 ± 0.02 (PNPDPP) and 47.0 ± 6.0 (DFP)) and V-type OPNA analogs (kcat/Km (M-1 min-1): 504.3 ± 48.5 (DSM) and 1324.0 ± 47.5 (malathion)). In conclusion, we developed GV-hybrid forms of rePON1 and bacterial OPH mutants as effective and suitable catalytic bioscavengers to hydrolyze a broad range of OPNA analogs.