• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1040-1043
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• 2008

Klebsiella oxytoca CCUG 15788 is resistant to $Ni^{2+}$ at a concentration of 10 mM and grows in an inducible manner when exposed to lower concentrations of $Ni^{2+}$. The complete genomic sequence of a 4.2-kb HindIII-digested fragment of this strain was determined from genomic DNA. It was shown to contain four nickel resistance genes (nirA, nirB, nirC, and nirD) encoding transporter and transmembrane proteins for nickel resistance. When the plasmid pKOHI4, encoding nirABCD, was transformed into Escherichia coli JM109, the cells were able to grow in Tris-buffered mineral medium containing 3 mM nickel. TnphoA'-1 insertion mutants in the four nickel genes nirA, nirB, nirC, and nirD showed nickel sensitivity. The nir genes were heterogeneously expressed in E. coli, suggesting functional roles of these genes in nickel resistance.

• Journal of Soil and Groundwater Environment
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• v.19 no.4
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• pp.62-69
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• 2014

In the results of monitoring nitrate concentration in more than 8,000 groundwater wells around agro-livestock, the average and maximum nitrate concentration was 9.4 mg/L and 101.2 mg/L, respectively. Since about 31% of the monitoring wells was exceed the quality standard for drinking water, nitrate control such as remediation or source regulation is required to conserve safe-groundwater in South Korea. Typical nitrate-treatment technologies include ion exchange, reverse osmosis, and biological denitrification. Among the treatment methods, biological denitrification by indigenous microorganism has environmental and economic advantages for the complete elimination of nitrate because of lower operating costs compared to other methods. Major mechanism of the process is microbial reduction of nitrate to nitrite and nitrogen gas. Three functional genes (nosZ, nirK, nirS) that encode for the enzyme involved in the pathway. In this work, we tried to develop simple process to determine possibility of natural denitrification reaction by monitoring the functional gene. For the work, the functional genes in nitrate-contaminated groundwater were monitored by using PCR with specific target primers. In the result, functional genes (nosZ and nirK) encoding denitrification enzymes were detected in the groundwater samples. This method can help to determine the possibility of natural-nitrate degradation in target groundwater wells without multiplex experimental process. In addition, for field-remediation application we selected nitrate-contaminated site where 200~600 mg/L of nitrate is continuously detected. To determine the possibility of nitrate-degradation by stimulated-natural attenuation, groundwater was sampled in two different wells of the site and nitrate concentration of the samples was 300 mg/L and 616 mg/L, respectively. Fumarate for different C/N ratio was added into microcosm bottles containing the groundwater to examine denitrification rate depending on carbon concentration. In the result, once 1.5 times more than amount of fumarate stoichiometry required was added, the 616 mg/L of nitrate and 300 mg/L of nitrate were completely degraded in 8 days and 30 days. The nitrite, byproduct of denitrification process, was also completely degraded during the experimental period.

• Environmental Engineering Research
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• v.20 no.1
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• pp.51-57
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• 2015

As expected, the expression of denitrifying genes in a Typha wetland (relatively stagnant compared to other ponds), showing higher nitrogen removal efficiency in summer, was affected by temperature. The abundance and gene transcripts of nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductase (norB), and nitrous oxide reductase (nosZ) genes in seasonal sediment samples taken from the Acorus and Typha ponds of free surface flow constructed wetlands were investigated using quantitative polymerase chain reaction (Q-PCR) and quantitative reverse transcription PCR (Q-RT-PCR). Denitrifying gene copy numbers ($10^5-10^8$ genes $g^{-1}$ sediment) were found to be higher than transcript numbers-($10^3-10^7$ transcripts $g^{-1}$ sediment) of the Acorus and Typha ponds, in both seasons. Transcript numbers of the four functional genes were significantly higher for Typha sediments, in the warm than in the cold season, potentially indicating greater bacterial activity, during the relatively warm season than the cold season. In contrast, copy numbers and expression of denitrifying genes of Acorus did not provide a strong correlation between the different seasons.

• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.72-78
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• 2020

The dynamics of nitrogen compounds and RNA-based functional genes were characterized in the nitrification-denitrification coupling process. For the removal of residual ammonium, intermittent aeration was introduced in the denitrification reactor. N₂O production was not observed in both reactors. In both reactors, the nitrifying genes (achaeal-amoA, bacterial-amoA and hor) and denitrifying genes (narG, nirK, norB and nosZ) had a copy number of 3.92 × 10²-7.25 × 10⁵ and 2.85 × 10²-3.06 × 10⁴ per ng of DNA, respectively. These results suggest that denitrification and nitrification reactions occur in both the nitrification and denitrification reactors, respectively. Therefore, the coupling process is a promising one for the conversion of ammonium to nitrogen without generating N₂O.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.7
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• pp.752-756
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• 2006

The independent anoxic reactor was introduced in biological aerated filters as the regulation of water quality requirement, especially total nitrogen, had been strengthened. The process studied in this work was upflow $Biobead^{(R)}$ process which was used commercial invented for removal of organic materials and nitrification. For the purpose of evaluating the independent anoxic reactor, PCR-DGGE, of the molecular biological methods, was performed. Two types of nitrite reductase genes were selected. One is nirS represented cytocrome $cd_1$ nitrite reductase gene and the other is nirK represented Cu-containing nitrite reductase gene. Denitrifier community in the independent anoxic reactor was analyzed with PCR-DGGE using these two denitrifying functional genes. As the result of the PCR, only nirS gene was detected between nirS and nirK. With the result of the DGGE, specific bands became strong, as the operating days were longer, nitrate loading rate was increased. otherwise those of the initial activated sludge showed various bands. In the consequence of the sequence of DGGE bands, various denitrifiers were sequenced in the initial activated sludge, while specific denitrifiers like alcaligenes faecalis were predominant in the anoxic reactor. Consequently, introduction of the independent anoxic reactor made it possible to achieve 96% denitrification efficiency, and was proper for the modification of BAF process.

• Journal of Soil and Groundwater Environment
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• v.20 no.7
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• pp.80-89
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• 2015

Nitrate is on the most seriou pollutant encountered in shallow groundwater aquifer in agricultural area. There are various remediation technologies such as ion exchange, reverse osmosis, and biological denitrification to recover from nitrate contamination. Biological denitrification by indigenous microorganism of the technologies has been reviewed and applied on nitrate contaminated groundwater. In this work, we selected the site where the annual nitrate (NO₃⁻) concentration is over 105 mg/L and evaluated denitrification process with sampled soil and groundwater from 3 monitoring wells (MW4, 5, 6). In the results, the nitrate degradation rate in each well (MW 4, 5, and 6) was 25 NO₃⁻ mg/L/day, 6 NO₃⁻ mg/L/day, and 3.4 NO₃⁻ mg/L/day, respectively. Nitrate degradation rate was higher in batch system treated with 2 times higher fumarate as carbon source than control batch system (0.42M fumrate/1M NO₃⁻), comparing with batch system with soil sample. This result indicates that increase of carbon source is more efficient to enhance denitrification rate than addition of soil sample to increase microbial dynamics. In this work, we also confirmed that monitoring method of functional genes (nirK and nosZ) involved in denitrification process can be applied to evaluated denitrifcation process possibility before application of field process such as in-situ denitrification by push-pull test.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.630-638
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• 2019

Nitrous oxide (N₂O) is a greenhouse gas with a global warming potential 310 times higher than that of carbon dioxide. In this study, an N₂O-reducing consortium was obtained by enrichment culture using advanced treatment sludge as the inoculum. The dominant bacteria in the consortium were Sulfurovum (17.95%), Geobacter (14.63%), Rectinema (11.45%), and Chlorobium (8.24%). The consortium displayed optimal N₂O reducing activity when acetate was supplied as the carbon source at a carbon/nitrogen ratio (mol·mol^-1) of 6.3. The N₂O reduction rate increased with increasing N₂O concentration at less than 3,000 ppm. Kinetic analysis revealed that the maximum N₂O reduction rate of the consortium was 163.9 ㎍-N·g-VSS^-1·h^-1. Genes present in the consortium included nosZ (reduction of nitrous oxide to N₂), narG (reduction of nitrate to nitrite), nirK (reduction of nitrite to nitric oxide), and norB (reduction of nitric oxide to nitrous oxide). These results indicate that the N₂O-reducing consortium is a promising bioresource that can be used in denitrification and N₂O mitigation.