• Microbiology and Biotechnology Letters
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• v.47 no.2
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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.

• 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.

• Journal of the Korean Chemical Society
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• v.57 no.3
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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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• v.35 no.12
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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%.

• Korean Journal of Microbiology
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• v.27 no.2
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• pp.139-146
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• 1989

The mechanisms and metabolic products involved in the degradation of an organophosphate insecticide, diazinon, were studied in submerged paddy soil under the laboratory condition at $30^{\circ}C$. Diazinon abatement in non-sterilized soil was more rapid than indicating microbial participation in diazinon in soil. One-half of the original applications was lost in 2 days and less than 5% remained after 7 days. During the same period, dizinon applications increased tha microbial populations in accordance with the monooxygenase and esterase activities in soil. These results suggest that the microbiological factors develop in soil following diazinon application. The esterase and monooxygenase-catalyzing degradation products of diazinon were isolated and tentatively identified by mass spectrometryas 2-isopropyle-6-methyl-4-hydroxy pyrimidine, diazoxon, hydroxydiazinon, and sulfotep.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.6
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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.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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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.

• Korean Journal of Agricultural Science
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• v.50 no.4
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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.