• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.717-724
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• 2016

Methane (CH₄) is the most abundant component in natural gas. To reduce its harmful environmental effect as a greenhouse gas, CH₄ can be utilized as a low-cost feed for the synthesis of methanol by methanotrophs. In this study, several methanotrophs were examined for their ability to produce methanol from CH₄; including Methylocella silvestris, Methylocystis bryophila, Methyloferula stellata, and Methylomonas methanica. Among these methanotrophs, M. bryophila exhibited the highest methanol production. The optimum process parameters aided in significant enhancement of methanol production up to 4.63 mM. Maximum methanol production was observed at pH 6.8, 30℃, 175 rpm, 100 mM phosphate buffer, 50 mM MgCl₂ as a methanol dehydrogenase inhibitor, 50% CH₄ concentration, 24 h of incubation, and 9 mg of dry cell mass ml^-1 inoculum load, respectively. Optimization of the process parameters, screening of methanol dehydrogenase inhibitors, and supplementation with formate resulted in significant improvements in methanol production using M. bryophila. This report suggests, for the first time, the potential of using M. bryophila for industrial methanol production from CH₄.

• 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 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.21 no.7
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• pp.753-756
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• 2011

A methane-oxidizing bacterium was isolated from the enriched culture of a landfill cover soil. The closest relative of the isolate, designated M6, is Methylocystis sp. Based on a kinetic analysis, the maximum specific methane oxidation rate and saturation constant were 4.93 mmol gdry cell $weight^{-1}{\cdot}h^{-1}$ and 23${\mu}M$, respectively. This was the first time a kinetic analysis was performed using pure methanotrophic culture. The methane oxidation by M6 was investigated in the presence of aromatic (m- and pxylene and ethylbenzene) or sulfur (hydrogen sulfide, dimethyl sulfide, methanthiol) compounds. The methane oxidation was inhibited by the presence of aromatic or sulfur compounds.

• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.244.2-244
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• 2001

No Abstract, See Full Text

• KSBB Journal
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• v.28 no.5
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• pp.275-281
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• 2013

Soil microbial community analysis of farmland soil sprayed with lye in order to use fertilizer in Nigeria was performed. As a control, two kinds of soils not sprayed with lye, located in Eungo and Lagos with general practice in agriculture was selected. Soil sprayed with lye was pH 8.25 through alkalization reaction, while the other soil samples were pH 6.22 and 5.94. Substrate utilization and species diversity index of soil sprayed with lye were low than that of the other soils with the analysis of Biolog ecoplate. As a result of principal component analysis, the relationship between three samples was low. Microbial community analysis was performed by DGGE and most of them were soil uncultured bacterium. Especially, Uncultured Acidobacteria and Uncultured Methylocystis sp., which had been isolated from the rhizosphere of soybean grown in that site were discovered in the soil sprayed with lye.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.76-81
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• 2012

Removal of methane, benzene and toluene was evaluated in a lab-scale biocover packed with a soil mixture of forest soil and earthworm cast (75:25 weight ratio). The bacterial community in the biocover was characterized using quantitative real-time PCR and terminal restriction fragment length polymorphism. Methane was removed at the upper layer of the biocover (-0.1 ~ -0.4 m), where the oxygen concentration was remarkably lower. The average removal efficiencies for methane and benzene/toluene were 90% and 99%, respectively. The pmoA gene copy numbers, responsible for methane oxidation, in the upper layer were higher than those in the lower layer. While type I methanotrohs dominated the lower layer, type II methanotrophs, such as Methylocystis and Methylosinus, were noted to be predominant in the upper layer. Benzene and toluene were removed from the lower layer (-0.6 ~ -0.9 m) as well as the upper layer. Moreover, the tmoA gene copy number, responsible for benzene/toluene oxidation, seen in the upper layer was not significantly different from those seen in the lower layer. These results suggest that a biocover packed with a soil and earthworm cast mixture is a promising method which could be utilized for the control of methane and volatile organic compounds such as benzene and toluene.

• Journal of Korean Society of Water Science and Technology
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• v.26 no.6
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• pp.27-42
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• 2018

This study analyzed the utility of ammonium chloride ($NH_4Cl$) as a nitrogen source for methanotroph communities. When cultured in nitrate mineral salt (NMS) medium, the methanotroph community we identified four families, seven genera, and 16 type I and type II species of methanotrophs. Among species in the Methylobacter genus, Methylobacter marinus could be actively cultured in NMS medium without NaCl addition. Following the addition of 25 mM $NH_4Cl$, the numbers of the type I genera Methylomonas, Methylococcus, and Methylobacter were increased, whereas the numbers of the type II genera Methylocystis and Methylosinus were decreased after 5 days. In methanotroph communities, certain concentrations of $NH_4Cl$ affected methane consumption and growth of methanotrophs at the community level. $NH_4Cl$ caused a considerable decrease in the methane consumption rate and the expression of soluble methane monooxygenases (sMMOs) but did not inhibit the growth of Methylomonas methanica expressing sMMO. These results could be attributed to competitive antagonism of MMOs due to their direct involvement in ammonia oxidation.

• 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.25 no.4
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• pp.423-430
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• 2015

Aerobic CH₄ oxidation is an important CH₄ sink in landfills. To investigate the distribution and community diversity of methanotrophs and link with soil characteristics and operational parameters (e.g., concentrations of O₂, CH₄), cover soil samples were collected at different locations and depths from the Mengzi semi-aerobic landfill (SAL) in Yunnan Province of southern China. Specific PCR followed by denaturing gradient gel electrophoresis and realtime PCR were used to examine methanotrophs in the landfill cover soils. The results showed that different locations did harbor distinct methanotroph communities. Methanotrophs were more abundant in areas near the venting pipes because of the higher O₂ concentrations. The depth of 20-25 cm, where the ratio of the CH₄ to O₂ was within the range from 1.3 to 8.6, was more conducive to the growth of CH₄-oxidizing bacteria. Type II methanotrophs dominated in all samples compared with Type I methanotrophs, as evidenced by the high ratio of Type II to Type I methanotrophic copy numbers (from 1.76 to 11.60). The total copy numbers of methanotrophs detected were similar to other ecosystems, although the CH₄ concentration was much higher in SAL cover soil. Methylobacter and Methylocystis were the most abundant Type I and Type II methanotrophs genera, respectively, in the Mengzi SAL. The results suggested that SALs could provide a special environment with both high concentrations of CH₄ and O₂ for methanotrophs, especially around the vertical venting pipes.