• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1706-1715
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• 2018

Several non-methylotrophic bacteria have been reported to improve the growth and activity of methanotrophs; however, their interactions remain to be elucidated. We investigated the interaction between Methylocystis sp. M6 and Microbacterium sp. NM2. A batch co-culture experiment showed that NM2 markedly increased the biomass and methane removal of M6. qPCR analysis revealed that NM2 enhanced both the growth and methane-monooxygenase gene expression of M6. A fed-batch experiment showed that co-culture was more efficient in removing methane than M6 alone (28.4 vs. $18.8{\mu}mol{\cdot}l^{-1}{\cdot}d^{-1}$), although the biomass levels were similar. A starvation experiment for 21 days showed that M6 population remained stable while NM2 population decreased by 66% in co-culture, but the results were opposite in pure cultures, indicating that M6 may cross-feed growth substrates from NM2. These results indicate that M6 apparently had no negative effect on NM2 when M6 actively proliferated with methane. Interestingly, a batch experiment involving a dialysis membrane indicates that physical proximity between NM2 and M6 is required for such biomass and methane removal enhancement. Collectively, the observed interaction is beneficial to the methanotroph but adversely affects the non-methylotroph; moreover, it requires physical proximity, suggesting a tight association between methanotrophs and non-methylotrophs in natural environments.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.5
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• pp.971-980
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• 2000

Mixed methanotrophs (MM) secreting soluble methane monooxygenase(sMMO) were immobilized on celite R-635 to degrade trichloroethylene(TCE) in methanotrophic consortium biofilm reactor(MCBR) system. Further neutralization of celite R-635 was not needed for immobilization because effluent pH was stabilized at neutral after 4 hour washing. It took 130 days to develop biofilm on celite R-635 and the color of the celite changed gradually from white to red. After biofilm developed, influent methane and oxygen were decreased from 2.5~4 and 8~10 ppm to 0.5~1 and 1~2 ppm, respectively, With influent 2 ppm of TCE and 10 hours of retention time, 79.9% of TCE was degraded in the MCBR system.

• Korean Journal of Environmental Agriculture
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• v.33 no.4
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• pp.306-313
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• 2014

BACKGROUND: Forest soils contain microbes capable of consuming atmospheric methane ($CH_4$), an amount matching the annual increase in $CH_4$ concentration in the atmosphere. However, the effect of plant residue production by different forest structure on $CH_4$ oxidation is not studied in Korea. The objective of this study was to evaluate the effect of Korean alpine soils having different forestation structure on $CH_4$ uptake rates. METHODS AND RESULTS: the $CH_4$ flux was measured at three sites dominated with pine, chestnut and oak trees in southern Korea. The $CH_4$ uptake potentials were evaluated by a closed chamber method for a year. The $CH_4$ uptake rate was the highest in the pine tree soil ($1.05mg/m^2/day$) and then followed by oak ($0.930mg/m^2/day$) and chestnut trees ($0.497mg/m^2/day$). The $CH_4$ uptake rates were highly correlated to soil organic matter and moisture contents, and total microbial and methanotrophs activities. Different with the general concent, there was no any correlation between $CH_4$ oxidation rates, and soil temperature and labile carbon concentrations, irrespective with tree species. CONCLUSION: Conclusively, the methane oxidation rate was correlated in positive manner with organic matter, abundance of methanotrophs. Methane oxidation was different among tree species. This results could be used to estimate methane oxidation rate in forest of Korea after complementing information about statistical data and methane oxidation of other site.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.152-156
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• 2012

Methanotrophic communities from freshwater wetland (FW), seawater wetland (SW), forest (FS), and landfill soils (LS) around Seoul of South Korea, were characterized using comparative sequence analyses of clone libraries. Proportions of Methylocaldum, Methlyococcus and Methylosinus were found to be greater in FW and SW, while Methylobacter and Methylomonas were more notable in FS and Methylocystis and Methylomicrobium more prominent in LS. Lag periods behind the initiation of methane oxidation significantly varied amongst the soils. Methane oxidation rates were greater in $FW{\geq}LS{\geq}SW>FS$ (p<0.05). Thus, the environmental setting is a significant factor influencing the communities and capabilities of methanotrophs.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2098-2105
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• 2016

Massive reserves of methane ($CH_4$) remain unexplored as a feedstock for the production of liquid fuels and chemicals, mainly because of the lack of economically suitable and sustainable strategies for selective oxidation of $CH_4$ to methanol. The present study demonstrates the bioconversion of $CH_4$ to methanol mediated by Type I methanotrophs, such as Methylomicrobium album and Methylomicrobium alcaliphilum. Furthermore, immobilization of a Type II methanotroph, Methylosinus sporium, was carried out using different encapsulation methods, employing sodium-alginate (Na-alginate) and silica gel. The encapsulated cells demonstrated higher stability for methanol production. The optimal pH, temperature, and agitation rate were determined to be pH 7.0, $30^{\circ}C$, and 175 rpm, respectively, using inoculum (1.5 mg of dry cell mass/ml) and 20% of $CH_4$ as a feed. Under these conditions, maximum methanol production (3.43 and 3.73 mM) by the encapsulated cells was recorded. Even after six cycles of reuse, the Na-alginate and silica gel encapsulated cells retained 61.8% and 51.6% of their initial efficiency for methanol production, respectively, in comparison with the efficiency of 11.5% observed in the case of free cells. These results suggest that encapsulation of methanotrophs is a promising approach to improve the stability of methanol production.

• Korean Journal of Environmental Agriculture
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• v.41 no.2
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• pp.115-124
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• 2022

BACKGROUND: Methane is a major greenhouse gas attributed to global warming partly contributed by agricultural activities from ruminant fermentation and rice paddy fields. Methanotrophs are microorganisms that utilize methane. Their unique metabolic lifestyle is enabled by enzymes known as methane monooxygenases (MMOs) catalyzing the oxidation of methane to methanol. Rice absorbs, transports, and releases methane directly from soil water to its stems and the micropores and stomata of the plant epidermis. Methylobacterium species associated with rice are dependent on their host for metabolic substrates including methane. METHODS AND RESULTS: Methylobacterium spp. isolated from rice were evaluated for methane oxidation activities and screened for the presence of sMMO mmoC genes. Qualitatively, the soluble methane monooxygenase (sMMO) activities of the selected strains of Methylobacterium spp. were confirmed by the naphthalene oxidation assay. Quantitatively, the sMMO activity ranged from 41.3 to 159.4 nmol min^-1 mg of protein^-1. PCR-based amplification and sequencing confirmed the presence and identity of 314 bp size fragment of the mmoC gene showing over 97% similarity to the CBMB27 mmoC gene indicating that Methylobacterium strains belong to a similar group. CONCLUSION(S): Selected Methylobacterium spp. contained the sMMO mmoC gene and possessed methane oxidation activity. As the putative methane oxidizing strains were isolated from rice and have PGP properties, they could be used to simultaneously reduce paddy field methane emission and promote rice growth.

• 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 Microbiology and Biotechnology
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• v.23 no.12
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• pp.1774-1778
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• 2013

Methylotrophs within biological activated carbon (BAC) systems have not received attention although they are a valuable biological resource for degradation of organic pollutants. In this study, methylotrophic populations were monitored for four consecutive seasons in BAC of an actual drinking water plant, using ribosomal tag pyrosequencing. Methylotrophs constituted up to 5.6% of the bacterial community, and the methanotrophs Methylosoma and Methylobacter were most abundant. Community comparison showed that the temperature was an important factor affecting community composition, since it had an impact on the growth of particular methylotrophic genera. These results demonstrated that BAC possesses a substantial methylotrophic activity and harbors the relevant microbes.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.221-227
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• 2013

The methane oxidation characteristics at the top and bottom layers in up-flow biofilters were investigated. Two biofilters were packed with perlite and tobermolite (biofilter A: respectively top and bottom; biofilter B: respectively bottom and top) and then compared. The methane oxidation rate was analyzed with the packed bed of the biofilter layers. The bacterial population in the biofilter was characterized using quantitative real-time PCR. For the methane oxidation rate of the biofilter A column, the perlite top part ($845.16{\pm}64.78{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$) gave a relatively higher value than the tobermolite bottom part ($381.85{\pm}42.00{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$). For the methane oxidation rate of the biofilter B column, the tobermolite top part ($601.25{\pm}37.78{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$) provided a relatively higher value than the perlite bottom part ($411.07{\pm}53.02{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$). The pmoA gene copy numbers, responsible for methanotrophs, in the top layer of biofilter A (1.27E+13 pmoA gene copy number/mg-VSS) was higher than in the bottom layer (3.33E+13 pmoA gene copy number/mg-VSS). However, the population of methanotrophs in biofilter B was not significantly different between the top and bottom layers. These results suggest that although the methane oxidation rates of perlite and tobermolite in the top parts of biofilter A and B were high, methanotroph populations were higher in the bottom parts of both biofilters, with a rapid decline in methane concentrations within the biofilters.

• Journal of Korean Society of Environmental Engineers
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• v.33 no.9
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• pp.662-669
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• 2011

This study was conducted to biologically convert methane into methanol. Methane contained in biogas was bio-catalytically oxidized by methane monooxygenase (MMO) of methanotrophs, while methanol conversion was observed by inhibiting methanol dehydrogenase (MDH) using MDH activity inhibitors such as phosphate, NaCl, $NH_4Cl$, and EDTA. The degree of methane oxidation by methanotrophs was the most highly accomplished as 0.56 mmol for the condition at $35^{\circ}C$ and pH 7 under 0.4 (v/v%) of biogas ($CH_4$ 50%, $CO_2$ 50%) / Air ratio. By the inhibition of 40 mM of phosphate, 50 mM of NaCl, 40 mM of $NH_4Cl$ and $150{\mu}m$ of EDTA, methane oxidation rate could achieve more than 80% regardless of type of inhibitors. In the meantime, addition of 40 mM of phosphate, 100 mM of NaCl, 40 mM of $NH_4Cl$ and $50{\mu}m$ of EDTA each led to generating the highest amount of methanol, i.e, 0.71, 0.60, 0.66, and 0.66 mmol when 1.3, 0.67, 0.74, and 1.3 mmol of methane was each concurrently consumed. At that time, methanol conversion rate was 54.7, 89.9, 89.6, and 47.8% respectively, and maximum methanol production rate was $7.4{\mu}mol/mg{\cdot}h$. From this, it was decided that the methanol production could be maximized as 89.9% when MDH activity was specifically inhibited into the typical level of 35% for the inhibitor of concern.