• Journal of Microbiology and Biotechnology
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• v.33 no.7
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• pp.886-894
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• 2023

During the rhizoremediation of diesel-contaminated soil, methane (CH₄), a representative greenhouse gas, is emitted as a result of anaerobic metabolism of diesel. The application of methantrophs is one of solutions for the mitigation CH₄ emissions during the rhizoremediation of diesel-contaminated soil. In this study, CH₄-oxidizing rhizobacteria, Methylocystis sp. JHTF4 and Methyloversatilis sp. JHM8, were isolated from rhizosphere soils of tall fescue and maize, respectively. The maximum CH₄ oxidation rates for the strains JHTF4 and JHM8 were 65.8 and 33.8 mmol·g-DCW^-1·h^-1, respectively. The isolates JHTF4 and JHM8 couldn't degrade diesel. The inoculation of the isolate JHTF4 or JHM8 significantly enhanced diesel removal during rhizoremediation of diesel-contaminated soil planted with maize for 63 days. Diesel removal in the tall fescue-planting soil was enhanced by inoculating the isolates until 50 days, while there was no significant difference in removal efficiency regardless of inoculation at day 63. In both the maize and tall fescue planting soils, the CH₄ oxidation potentials of the inoculated soils were significantly higher than the potentials of the non-inoculated soils. In addition, the gene copy numbers of pmoA, responsible for CH₄ oxidation, in the inoculated soils were significantly higher than those in the non-inoculated soils. The gene copy numbers ratio of pmoA to 16S rDNA (the ratio of methanotrophs to total bacteria) in soil increased during rhizoremediation. These results indicate that the inoculation of Methylocystis sp. JHTF4 and Methyloversatilis sp. JHM8, is a promising strategy to minimize CH₄ emissions during the rhizoremediation of diesel-contaminated soil using maize or tall fescue.

• Journal of Soil and Groundwater Environment
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• v.24 no.5
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• pp.42-54
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• 2019

Biodegradation of an explosive compound, glyceryl trinitrate (GTN), was studied with a denitrifying microbial culture grown in a sequencing batch reactor and a GTN acclimated denitrifying culture. The GTN acclimated culture, which were fed on GTN for 1 month, degraded GTN regioselectively via denitration on C1 position as compared to C2 position denitration by denitrifying culture that has never been exposed to GTN. Accumulation of two isomeric glyceryl dinitrates (GDNs) in both culture medium suggests that GDN denitration is the rate-limiting step in GTN biodegradation. The first order GTN degradation rate normalized to cell concentration of the acclimated culture was calculated to be 0.045 (${\pm}0.002$) L/g-hr. Increasing concentration of electron acceptor(nitrate) resulted in discouraged GTN degradation. According to microbial community analysis, prolonged GTN exposure resulted in 25% increase in the genus level of the GTN acclimated culture with the disappearance of two dominating denitrifying microbial species of Methyloversatilis universalis and Hyphomicrobium zavarzinii in the denitrifying culture.

• Journal of Korean Society on Water Environment
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• v.36 no.6
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• pp.581-591
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• 2020

In this study, two types of reactors were operated to examine the properties of methanol uptake under the high-rate denitrification process. In a sequencing batch reactor, the denitrifying activity was enriched up to 0.80 g-N/g-VSS-day for 72 days. Then, the enriched denitrifying sludge was transferred to a completely stirred tank reactor (CSTR). At the final phase on Day 46-50, the nitrogen removal efficiency was around 100% and the total nitrogen removal rate reached 0.097±0.003 kg-N/㎥-day. During the continuous process, the sludge settling index (SVI30) was stabilized as 118.3 mL/g with the biomass concentration of 1,607 mg/L. The continuous denitrifying process was accelerated by using a sequencing batch reactor (SBR) with a total nitrogen removal rate of 0.403±0.029 kg-N/㎥-day with a high biomass concentration of 8,433 mg-VSS/L. Because the reactor was open to ambient air with the dissolved oxygen range of 0.2-0.5 mg-O2/L, an increased organic carbon requirement of 5.58±0.70 COD/NO3--N was shown for the SBR in comparison to the value of 4.13±0.94 for the test of the same biomass in a completely anaerobic batch reactor. The molecular analysis based on the 16S rRNA gene showed that Methyloversatilis discipulorum and Hyphomicrobium zavarzinii were the responsible denitrifiers with the sole organic carbon source of methanol.

• Journal of Korean Society on Water Environment
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• v.39 no.1
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• pp.61-75
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• 2023

Denitrifying bacteria convert nitrate to nitrogen gas using an external carbon source as an electron donor. The external carbon source affects the denitrification performance and bacterial community structure. Although methanol is a cheap and effective external carbon source, the addition of diverse carbon sources may improve the total nitrogen removal rate and biomass characteristics, such as settleability. In this study, denitrifying reactions were performed using solely methanol and mixed carbon sources of methanol, glucose, and acetate in a sequencing batch reactor. The denitrifying reactor using methanol resulted in a total nitrogen removal rate of 0.39 ± 0.025 kg-N/m³-day while the suspended biomass transformed into dark brown granules. Methyloversatilis discipulorum had the highest predominance at 43.84%. The individual denitrifying biomasses, which were separately enriched with methanol, glucose, and acetate, showed the same total nitrogen removal performance of 0.39 ± 0.016 kg-N/m³-day. However, the addition of mixed carbon sources showed an improved total nitrogen removal rate of 0.42 ± 0.043 kg-N/m³-day, with the domination of Candidatus Saccaribacteria at 25.61%. The denitrifying granules turned pale yellow color. Influent COD/NO₃^--N ratios of 3.5, 5, and 7.5 exhibited COD/NO₃^--N consumptions of 4.3 ± 0.4, 4.4 ± 0.8, and 5.2 ± 0.7, and the consistent predominance of Candidatus Saccharibacteria.