• 한국미생물·생명공학회지
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• 제47권2호
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• pp.278-288
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• 2019

In the present investigation, we attempted to design a protocol to develop a hybrid protein with better bioremediation capacity. Using in silico approaches, a Hybrid Open Reading Frame (Hybrid ORF) is developed targeting the genes of microorganisms known for degradation of organophosphates. Out of 21 genes identified through BLAST search, 8 structurally similar genes (opdA, opd, opaA, pte RO, pdeA, parC, mpd and phnE) involved in biodegradation were screened. Gene conservational analysis categorizes these organophosphates degrading 8 genes into 4 super families i.e., Metallo-dependent hydrolases, Lactamase B, MPP and TM_PBP2 superfamily. Hybrid protein structure was modeled using multi-template homology modeling (3S07_A; 99%, 1P9E_A; 98%, 2ZO9_B; 33%, 2DXL_A; 33%) by $Schr{\ddot{o}}dinger$ software suit version 10.4.018. Structural verification of protein models was done using Ramachandran plot, it was showing 96.0% residue in the favored region, which was verified using RAMPAGE. The phosphotriesterase protein was showing the highest structural similarity with hybrid protein having raw score 984. The 5 binding sites of hybrid protein were identified through binding site prediction. The docking study shows that hybrid protein potentially interacts with 10 different organophosphates. The study results indicate that the hybrid protein designed has the capability of degrading a wide range of organophosphate compounds.

• 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.

• 대한화학회지
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• 제57권3호
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• pp.382-388
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• 2013

The present paper focuses on the synthesis, characterization and application of nanophotocatalyst for degradation of quinalphos and monocrotophos. Novel heterostructured ZnO/$TiO_2$ photocatalyst ($Z_9T$) was prepared and characterized with X-ray diffraction (XRD), SEM and UV-vis diffuses reflectance spectroscopy. The average crystalline size of synthesized $Z_9T$ was found to be 21.48 nm. The pesticides were degraded in the presence of nanophotocatalysts i.e., $TiO_2$, ZnO, $TiO_2$/ZnO mixed in various proportions and heterostructured nanophotocatalyst synthesized by Sol-Gel method. The batch experiments were performed by adding photocatalyst to 100 ml of pesticide solution and suspension was subjected to irradiation under UV light. In case of mixed catalyst, the maximum degradation of monocrotophos and quinalphos has been observed when ZnO and $TiO_2$ were in the ratio of 7:3 and 8:2 respectively. The degradation efficiency with synthesized heterostructured nanophotocatalyst ($Z_9T$) was found to be comparable with $TiO_2$.

• Bulletin of the Korean Chemical Society
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• 제35권12호
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• pp.3521-3526
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• 2014

A magnetic sonophotocatalyst $Fe_3O_4@SiO_2@TiO_2$ is synthesized for the enhanced biodegradability of organophosphate pesticide. The as-prepared catalysts were characterized using different techniques, such as X-ray diffraction (XRD) and transmission electron microscopy (TEM). The radial sonophotocatalytic activity of $Fe_3O_4@SiO_2@TiO_2$ nanocomposite was investigated, in which commercial dichlorvos (DDVP) was chosen as an object. The degradation efficiency was evaluated in terms of chemical oxygen demand (COD) and enhancement of biodegradability. The effect of different factors, such as reaction time, pH, the added amount of catalyst on $COD_{Cr}$ removal efficiency were investigated. The average $COD_{Cr}$ removal efficiency reached 63.13% after 240 min in 12 L sonophotocatalytic reactor (catalyst $0.2gL^{-1}$, pH 7.3). The synergistic effect occurs in the combined sonolysis and photocatalysis which is proved by the significant improvement in $COD_{Cr}$ removal efficiency compared with that of solo photocatalysis. Under this experimental condition, the $BOD_5/COD_{Cr}$ ratio rose from 0.131 to 0.411, showing a remarkable improvement in biodegradability. These results showed that sonophotocatalysis may be applied as pre-treatment of pesticide wastewater, and then for biological treatment. The synthesized magnetic nanocomposite had good photocatalytic performance and stability, as when it was used for the fifth time, the $COD_{Cr}$ removal efficiency was still about 62.38%.

• 미생물학회지
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• 제27권2호
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• pp.139-146
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• 1989

담수상태의 논토양에 처리한 유기인계 살충제 diazinon의분해기작과 그 분해산물에 대하여 조사하였다. 항온조건($30^{\circ}C$)하에서 살균토양에 비하여 비살균토양에서의 diazinon 분해가 보다 빠르게 일어남으로써 담수토양내 diazinon 분해에 미생물의 대사활동이 매우 종요한 것으로 나타났는데, 비살균토양내 diazinon의 반감기는 약 2일이엇고 항온반응 7일만에 95% 이상의 분해가 이루어졌다. 또한 diazinon 분해기간 중 미생물 개체수와 아울러 토양내 monoxygenase 및 esterase 활성의 증가가 이루어졌으며 이는 토양내 diazinon 처리가 이 약제의 분해와 관련된 미생물학적 요인의 생장 또는 활성을 촉진시키고 있음을 보여준다. 이와 관련하여 담수토양내 diazinon 분해산물로서 esterase 작용에 의한 2-isopropyl-6-methyl-4-hydroxy pyrimidine 이외에 monooxygenase에 의한 diazoxon과 hydroxydiazinon, 그리고 sulfotep 등이 mass spectrometry에 의하여 확인되었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권6호
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• pp.436-440
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• 2000

Recently, we reported an improved technology for the degradation of organophosphate nerve agents using whole cells of genetically engineered Escherichia coli that anchored and displayed the enzyme organophosphorus hydrolase on the cell surface. In this paper we report the immobilization of these cells on highly porous sintered glass beads and the subsequent application of the immobilized cell in a continuous-flow packed bed bioreactor for the biodetoxification of a widely used insecticide, coumaphos.

• 한국미생물·생명공학회지
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• 제52권1호
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• pp.44-54
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• 2024

One of the serious modern environmental problems is pollution caused by highly toxic pesticides. Only small amounts of applied pesticides reach their target, and the rest ends up in soil and water. Chlorpyrifos is a toxic, broad-spectrum organophosphate insecticide. In humans, chlorpyrifos inhibits acetylcholinesterase (AChE) in the peripheral and central nervous system, and particularly in children, small amounts of this pesticide cause neurotoxic damage. As the toxic effects of chlorpyrifos and its persistence in the environment require its removal from contaminated sites, it is essential to study the biological diversity of chlorpyrifos-degrading microorganisms. In this study, we sought to determine the chlorpyrifos-degrading ability of the bacterial strain Ochrobactrum intermedium PDB-3. This strain was isolated from soil contaminated with various pesticides and identified as PDB-3 based on morpho-cultural characteristics, MALDI-TOF MS, and 16S rRNA. Studies were conducted for 30 days in sterile soils containing initial concentrations of 50, 75, 100, and 125 mg/kg of chlorpyrifos. To determine the degradation of chlorpyrifos, a liquid culture of the strain was added to the soil at three optical densities: 0, and after 24 and 48 h (OD = 0.03, 0.2 and 0.32). Using GX-MS, we determined that chlorpyrifos was converted to 3,5,6-trichloro-2-pyridinol (TCP). We also found that with increasing optical density, rapid degradation of the initial concentration of chlorpyrifos occurred. Sterile soil without strain PDB-3 was used as a control sample.

• Journal of Microbiology and Biotechnology
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• 제19권2호
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• pp.113-120
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• 2009

Twenty-seven fenitrothion-degrading bacteria were isolated from different soils, and their genetic and phenotypic characteristics were investigated. Analysis of the 16S rDNA sequence showed that the isolates were related to members of the genera Burkholderia, Pseudomonas, Sphingomonas, Cupriavidus, Corynebacterium, and Arthrobacter. Among the 27 isolates, 12 different chromosomal DNA fingerprinting patterns were obtained by polymerase chain reaction(PCR) amplification of repetitive extra genic palindromic(REP) sequences. The isolates were able to utilize fenitrothion as a sole source of carbon and energy, producing 3-methyl-4-nitrophenol as the intermediate metabolite during the complete degradation of fenitrothion. Twenty-two of 27 isolates were able to degrade parathion, methyl-parathion, and p-nitrophenol but only strain BS2 could degrade EPN(O-ethyl-O-p-nitrophenyl phenylphosphorothioate) as a sole source of carbon and energy for growth. Eighteen of the 27 isolates had plasmids. When analyzed with PCR amplification and dot-blotting hybridization using various specific primers targeted to the organophosphorus pesticide hydrolase genes of the previously reported isolates, none of the isolates showed positive signals, suggesting that the corresponding genes of our isolates had no significant sequence homology with those of the previously isolated organophosphate pesticide-degrading bacteria.

• 농업과학연구
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• 제50권4호
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• pp.941-952
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• 2023

The metabolites of agrichemicals, such as organophosphorus pesticides, are known to be more hazardous than their parent pesticides. 3,5,6-trichloro-2-pyridinol (TCP) is a major degradation product of chlorpyrifos, one of the organophosphate insecticides widely used in agriculture. In vivo or in vitro exposure to chlorpyrifos has been known to interfere with male reproductive functions, leading to reduced fertility in mammals. Therefore, this study was performed to examine the changes in the fertilization competence of boar spermatozoa exposed to TCP. Sperm samples were subjected to varying concentrations of TCP (10, 50, 100, 200 µM) and different periods of incubation. Sperm motility, motion kinematics, viability, acrosome integrity, intracellular reactive oxygen species (ROS) production, and gene expression levels (ODf2, ZPBP2, AKAP3 and AKAP4) were evaluated after exposure of the sperm to TCP. A significant dose-dependent reduction in motility was observed in sperm samples incubated with TCP compared to the controls after both incubation periods. Sperm viability was significantly decreased in samples incubated with 50, 100, and 200 µM TCP in both incubation periods. A significantly lower percentage of normal acrosomes and gene expression levels were observed in sperm samples exposed to 50, 100, and 200 µM TCP after both incubation periods, compared to the controls. There was a significant increase in the ROS production in spermatozoa incubated with 100 - 200 µM TCP after both incubation periods. Consequently, the direct exposure of boar spermatozoa to TCP interferes with sperm functions and leads to decreased fertilization. In order to identify and address the various causes of reproductive decline, the impact of chemical metabolites needs to be discussed in depth.