• Korean Journal of Microbiology
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• v.37 no.1
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• pp.66-71
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• 2001

An aerobic microbiological process was tested in 1.5 L stirred tank reactors for the treatment of dinitrotoluenes (DNTs)[e.g., 2.4-dinitrotoluene (2,4-DNT), 2,6-dinitrotoluene (2,6-DNT)] in the test culture of Stenotrophomonas maltophilia, which had previously characterized. Both 2,4-DNT and 2,6-DNT were completely degraded within 10 days and 14 days of incubation, respectively. Addition of the secondary carbon was essential to degrade DNTs, and little degradation was achieved in the absence of the secondary carbons. The effect of additional nitrogens on the degradation of DNTs was evaluated. Complete degradation of DNTs was observed in the absence of any additional nitrogens, whereas DNTs were partially degraded in the growth media with additional nitrogens. Both HPLC and GC-MS were used to detect and verify the residual DNTs and their intermediates. As the results, the HPLC and GC-MS chromatograms demonstrated that the both parent compounds, 2, 4-DNT and 2,6-DNT, and respective intermediates, 2-amino-4-nitrotoluene and 2-amino-6-nitrotoluene, could be successfully identified under the analytical conditions.

• Journal of Environmental Science International
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• v.15 no.11
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• pp.997-1002
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• 2006

This report presents a voltammetric assay of dinitrotoluene using a DNA immobilized onto a carbon nanotube paste electrode (PE). The cyclic voltammetry (CV) and square wave (SW) stripping voltammetry parameters of the optimized conditions were obtained. An anodic peak current appeared at 0.3 V (versus Ag/AgCl) in a 0.1-M $NH_4H_2PO_4$ electrolyte solution. The detection limit was found to be $0.6ngL^{-1}$(S/N = 10), within a deposition time of 100 sec.

• Mycobiology
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• v.37 no.1
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• pp.17-20
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• 2009

White-rot fungus Irpex lacteus degraded TNT significantly in proportion to the culture time. After 48 h incubation, about 95% of TNT was degraded. Two reduced metabolites were identified as 4-amino-2,6-dinitrotoluene (4-ADNT) and 2-amino-4,6-dinitrotoluene (2-ADNT) which was further degraded.

• KSBB Journal
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• v.24 no.6
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• pp.556-562
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• 2009

The biological treatment of TNT-containing wastewater, known commonly as pink water, was investigated using a stirred tank reactor with Stenotrophomonas maltophilia OK-5 bacterial culture. S. maltophilia OK-5 exhibited effective degradation of TNT contained in pink water, completely degrading TNT (100 mg/L) within 6 days of incubation. The dark-red brown color derived from Hydride-Meisenheimer complex became more pronounced during the incubation period, which was determined quantitatively. High-pressure liquid chromatography was used to measure residual TNT, which also resolved the metabolic intermediates (i.e., 2,4-dinitrotoluene, 2,6-dinitrotoluene and 2,4-dinitro-6-hydroxytoluene). Gas chromatography-mass spectrometry was used to verify these intermediates. Quantification of the nitroreductase (pnrB) gene isolated from S. maltophilia OK-5 growing in pink water was performed with real-time PCR. The amount of pnrB gene copies increased to $10^3$-fold after 5 days of incubation time.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2001.12a
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• pp.96-96
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• 2001

• Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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• 2009.10a
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• pp.403-404
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• 2009

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.2
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• pp.105-111
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• 2002

The biological removal of 2,4,6-trinitrotoluene (TNT) was studied in a bench-scale bioreactor using a bacterial culture of strain OK-5 originally Isolated from soil samples contaminated with TNT. The TNT was completely removed within 4 days of incubation in a 2.5 L bench-scale bioreactor containing a newly developed medium. The TNT was catabolized in the presence of different supplemented carbons. Only minimal growth was observed in the killed controls and cultures that only received TNT during the incubation period. This catabolism was affected by the concentration ratio of the substrate to the biomass. The addition of various nitrogen sources produced a delayed effect for the TNT degradation. Tween 80 enhanced the degradation of TNT under these conditions. Two metabolic intermediates were detected and identified as 2-amino-4, 6-dinitrotoluene and 4-amino-2, 6-dinitrotoluene based on HPLC and GC-MS analyses, respectively. Strain OK-5 was characterized using the BIOLOG system and fatty acid profile produced by a microbial identification system equipped with a Hewlett Packard HP 5890 II gas chromatograph. As such, the bacterium was identified as a Stenotrophomonas species and designated as Stenotrophomonas sp. OK-5.

• Bulletin of the Korean Chemical Society
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• v.31 no.12
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• pp.3703-3706
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• 2010

Biomolecules specific to explosives can be exploited as chemical sensors. Peptides specific to immobilized dinitrotoluene (DNT) were identified using a phage display library. A derivative of DNT that contained an extended amine group, 4-(2,4-dinitrophenyl)butan-1-amine, was synthesized and immobilized using a self-assembled monolayer surface on gold. Filamentous M13 phages displaying random sequences of 12-mer peptides specific to the immobilized DNT-derivate were isolated from the M13 phage library by biopanning. A common peptide sequence was identified from the isolated phages and the synthesized peptides showed selective binding to DNT. When the peptide was immobilized on a quartz crystal microbalance (QCM) chip, it showed a binding signal to DNT, while toluene barely showed significant binding to the QCM chip. These results demonstrate that peptides screened by biopanning against immobilized DNT can be useful for quick and accurate detection of DNT.

• Bulletin of the Korean Chemical Society
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• v.34 no.2
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• pp.460-464
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• 2013

Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with $K_D$ value of $6.0{\times}10^{-7}$ M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.2
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• pp.198-205
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• 2005

This study was carried out to investigate the removal of TNT (2,4,6-trinitrotoluene) in the batch and continuous type microcosm systems consisting of marsh and pond. First, the batch reactor study showed that TNT (10 mg/L) was completely removed in the marsh and pond system within 20 days. The major reductive metabolites of TNT include 4-amino-2,6-dinitrotoluene (4-ADNT), 2-amino-4,6-dinitrotoluene (2-ADNT), 2,4-diamino-6-nitrotoluene (2,4-DANT), and 2,6-diamino-4-nitrotoluene (2,6-DANT). These metabolites concentration also decreased during further treatment. The continuous reactor systems combining marsh and pond indicated the similar pattern of TNT degadation and the metabolites production. Among the continuous reactor combinations, marsh-pond system showed more stable TNT removal and metabolites production. The toxicity of the effluent from the continuous system was examined by Microtox Assay using Vibrio fischeri. The result showed that the effluent toxicity was reduced below toxicity endpoint ($EC_{50}$) after continuous marsh pond system, indicating that metabolites of TNT are less toxic than TNT itself. Based on the results, TNT contaminated wastewater can be efficiently treated using marsh and pond wetland systems.