• Journal of Microbiology and Biotechnology
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• 제28권10호
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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.

• 대한환경공학회지
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• 제22권5호
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• pp.971-980
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• 2000

가용성 메탄산화효소를 분비하는 혼합 메탄자화균총을 celite R-635에 고정화시켜 TCE 분해를 위한 새로운 형식의 가압 산기식 혼합 메탄자화균총 고정층 생물막 반응기를 설계하였다. Celite R-635에서 용출되는 용액의 pH는 약 4시간 후부터 안정화되어서 중성 영역에 도달하므로 더 이상 중화할 필요가 없었다. 혼합 메탄자화균 생물막을 완전히 형성하기 위해서는 130일이 걸렸으며, 처음에 흰색을 띠고 있었던 celite는 점차 붉게 변해 갔었다. 생물막이 형성된 후에는 메탄과 산소를 각각 2.5~4, 8~10 ppm씩 공급할 때 하루 동안 체류한 후 0.5~1, 1~2 ppm 정도로 농도가 낮아졌다. 초기에 2 ppm의 TCE를 메탄자화균 고정층 생물막 반응기에서 10시간 동안 체류시켰을 때 79.9%의 분해 효율을 보였다.

• 한국환경농학회지
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• 제33권4호
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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.

• 한국미생물·생명공학회지
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• 제40권2호
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• pp.152-156
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• 2012

서울 근교의 민물 습지(FW), 해수습지(SW), 산림 토양(FS) 그리고 매립지 복토(LS)의 메탄산화세균 군집을 clone library/sequencing 기법을 이용하여 분석하였다. 메탄산화세균인 Methylocaldum, Methlyococcus과 Methylosinus는 FS와 SW에서 풍부하였으며, Methylobacter와 Methylomonas는 FW에서 풍부하였고, Methylocystis와 Methylomicrobium은 LS에서 우점하였다. 메탄 산화가 관찰되기 전까지 필요한 lag phase는 각 토양별로 유의적으로 차이가 있었고, 메탄 산화속도는 $FW{\geq}LS{\geq}SW>FS$순이었다. 이러한 결과들은 토양의 환경조건은 메탄산화세균의 군집과 메탄산화능에 영향을 미치는 중요한 인자임을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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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.

• 한국환경농학회지
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• 제41권2호
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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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• 제33권7호
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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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• 제23권12호
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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.

• 한국미생물·생명공학회지
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• 제41권2호
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• pp.221-227
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• 2013

Perlite와 tobermolite 담체를 각각 상 하단(바이오필터 A) 또는 하 상단(바이오필터 B)으로 충전한 상향식 바이오필터에서 상하단의 메탄산화 특성을 비교하였다. 각 바이오필터에서 상단과 하단 담체를 채취하여 메탄산화속도를 측정하고, 정량적 real time PCR 방법을 이용하여 메탄산화세균수를 정량분석 하였다. 혼합담체의 층적배열 차이에 따른 각 담체의 순수 메탄산화속도를 조사한 결과, biofilter A perlite 상단이 $845.16{\pm}64.78{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$로 tobermolite 하단 $381.85{\pm}42.00{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$에 비하여 상대적으로 높았다(p < 0.005). 또한, biofilter B tobermolite의 상단($601.25{\pm}37.78{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$)이 perlite 하단($411.07{\pm}53.02{\mu}mol{\cdot}VS^{-1}{\cdot}h^{-1}$)에 비하여 메탄산화속도가 높았다(p < 0.005). 바이오필터 A는 상단(1.27E+13 pmoA gene copy number/mg-VSS)보다는 하단(3.33E+13 pmoA gene copy number/mg-VSS) (p < 0.05)에 더 많은 메탄 산화 세균이 존재하였다. 그러나, 바이오필터 B는 상단과 하단간의 메탄 산화 세균수의 차이는 유의하지 않은 것으로 나타났다(p > 0.05). Perlite와 tobermolite로 충진된 각 담체의 순수 메탄산화속도는 상단부에 위치한 담체들이 높았음에도 불구하고 바이오필터내에서 메탄농도 감소폭이 컸던 하단부에서 메탄산화세균이 많이 존재하였다.

• 대한환경공학회지
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• 제33권9호
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• pp.662-669
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• 2011

본 연구에서는 메탄의 생물학적 메탄올 전환에 관한 연구를 수행하였다. 바이오가스 중의 메탄은 메탄산화균의 methane monooxygenase (MMO)의 생물학적 촉매반응에 의해 산화되었으며, 인산염, NaCl, $NH_4Cl$, EDTA와 같은 methanol dehydrogenase (MDH)의 활성 저해제를 이용하여 MDH의 활성도를 저해함으로써 메탄올의 전환이 이루어졌다. 메탄산화균은 $35^{\circ}C$, pH 7, 인공 바이오가스($CH_4$ 50%, $CO_2$ 50%) / Air의 부피비가 0.4인 조건에서 메탄 산화 정도가 0.56 mmol로 최대로 나타났다. 인산염 40 mM, NaCl 50 mM, $NH_4Cl$ 40 mM, EDTA $150{\mu}m$ 이하일 때 저해제의 종류에 상관없이 메탄 산화율은 80% 이상을 달성하였다. 한편, 인산염 40 mM, NaCl 100 mM, $NH_4Cl$ 40 mM, EDTA $50{\mu}m$ 주입 시 각각 1.30, 0.67, 0.74, 1.30 mmol의 메탄이 산화되는 동시에 각각 0.71, 0.60, 0.66, 0.66 mmol의 메탄올이 최대로 생성되었다. 이때의 메탄올 전환율은 각각 54.7, 89.9, 89.6 및 47.8%였으며 최대 메탄올 생성 속도는 $7.4{\mu}mol/mg{\cdot}h$였다. 이로부터 대상 저해제로 MDH 활성도를 일반적으로 35% 저해 시에 메탄올 생산량이 최대인 89.9%까지 나타남을 알 수 있었다.