• KSBB Journal
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• v.13 no.5
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• pp.483-490
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• 1998

Two unidentified methanotrophic strains (MM-white and MM-red) secreting soluble methane monooxygenase (sMMO) involved in thrichloroethylene biodegradation have been isolated from mixed methanotrophic consortium (MM) around Taejon area. Subsequently four methanotrophic strains were isolated from MM and named according to their color; white (MS-white), yellow (MS-yellow), pink (MS-pink) and reddish brown (MS-rbrown). All strains except MS-yellow which can take glucose as well as methane, metabolized methane as a sole carbon source. They all showed symbiotic behavior when methane was used as the sole carbon source. Optimum conditions of cell growth for MM were pH of 6.8 - 7.2, temperature of 29 - 32$^{\circ}C$, and gas flow rate of 6 (for methane), 40 (for air), and 4 ml/min (for carbon dioxide). The sMMO activity was expressed as naphthalene oxidation rate (${\mu}$mol/ mg protein/ hr). The sMMO activity for MM grown in flask culture with 1 ${\mu}$M of CuSO4 was 36, while it was 61 without copper. The activity for MM grown in the fermentor without CuSO4 was 1077, but is was 197 after reaction with 5 ppm of TCE. The methanotrophs showed significantly high sMMO activity despite the presence of 1 ${\mu}$M of CuSO4, although most of other strains already known could not express sMMO activity under this condition.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.134-139
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• 2006

Methanotrophs are microorganisms that possess the unique ability to utilize methane as their sole source of carbon and energy. A novel culture system, known as the compulsory circulation diffusion system, was developed for rapid growth of methanotrophic bacteria. Methanol dehydrogenase (MDH, EC 1.1.99.8) from Methylomicrobium sp. HG-1, which belongs to the type I group of methanotrophic bacteria, can catalyze the oxidation of methanol directly into formaldehyde. This enzyme was purified 8-fold to electrophoretic homogeneity by means of a 4 step procedure and was found in the soluble fraction. The relative molecular weight of the native enzyme was estimated by gel filtration to be 120 kDa. The enzyme consisted of two identical dimers which, in turn, consisted of large and small subunits in an ${\alpha}_2{\beta}_2$ conformation. The isoelectric point was 5.4. The enzymatic activity of purified MDH was optimum at pH 9.0 and $60^{\circ}C$, and remained stable at that temperature for 20 min. MDH was able to oxidize primary alcohols from methanol to octanol and formaldehyde.

• 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.

• New & Renewable Energy
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• v.19 no.4
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• pp.46-52
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• 2023

Methane is the second most emitted greenhouse gas after carbon dioxide. Despite lower emissions than those of carbon dioxide, methane receives significant attention owing to its more than 20-fold higher global warming potential. Consequently, the importance of research on methanotrophic bacteria, microorganisms capable of converting methane gas into high-value materials, is increasingly emphasized. In the case of methanotrophic bacteria, knowledge on episomal plasmids that can be used for genetic engineering remains lacking, which poses significant challenges to the engineering process. The replication origin sequences of natural plasmids within methanotrophic bacteria have been predicted through in silico methods. The basic characteristics of the replication origin, such as a high A/T ratio, repetitive sequences, and proximity to proteins related to replication, have been used as criteria for identifying the replication origin. As a result, a region with a sequence of 18 base pairs repeated eight times could be identified. The putative replication origin sequence thus identified generally takes the form of iterons, but it also possesses unique features such as the length of the gap between iterons and the repetition of identical iteron sequences. This information can be valuable for future design of episomal plasmids applicable to methanotrophs.

• Environmental Engineering Research
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• v.19 no.1
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• pp.75-81
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• 2014

Methanotrophic denitrification under a non-aerated condition (without external supply of oxygen or air) was investigated in a bioreactor coupled with a membrane diffuser. Batch experiment demonstrated that both methane consumption and nitrogen production rates were not high in the absence of oxygen, but most of the nitrate was reduced into $N_2$ with 88% recovery efficiency. The methane utilized for nitrate reduction was determined at 1.63 mmol $CH_4$/mmol $NO_3{^-}$-N, which was 2.6 times higher than the theoretical value. In spite of no oxygen supply, methanotrophic denitrification was well performed in the bioreactor, due to enhanced mass transfer of the methane by the membrane diffuser and utilization of oxygen remaining in the influent. The denitrification efficiency and specific denitrification rate were 47% and 1.69 mg $NO_3{^-}-N/g\;VSS{\cdot}hr$, respectively, which were slightly lower than for methanotrophic denitrification under an aerobic condition. The average concentration of total organic carbon in the effluent was as low as 2.45 mg/L, which indicates that it can be applicable as a post-denitrification method for the reclamation of secondary wastewater effluent. The dominant fatty acid methyl ester of mixed culture in the bioreactor was $C_{16:1{\omega}7c}$ and $C_{18:1{\omega}7c}$, which was predominantly found in type I and II methanotrophs, respectively. This study presents the potential of methanotrophic denitrification without externally excess oxygen supply as a post-denitrification option for various water treatment or reclamation.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1597-1605
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• 2014

Methane is considered as a next-generation carbon feedstock owing to the vast reserves of natural and shale gas. Methane can be converted to methanol by various methods, which in turn can be used as a starting chemical for the production of value-added chemicals using existing chemical conversion processes. Methane monooxygenase is the key enzyme that catalyzes the addition of oxygen to methane. Methanotrophic bacteria can transform methane to methanol by inhibiting methanol dehydrogenase. In this paper, we review the recent progress made on the biocatalytic conversion of methane to methanol as a key step for methane-based refinery systems and discuss future prospects for this technology.

• Membrane and Water Treatment
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• v.8 no.2
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• pp.171-184
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• 2017

This study presents the effect of hydraulic retention time (HRT) on the characteristics of emission of three major greenhouse gases (GHGs) including $CH_4$, $CO_2$ and $N_2O$ during operation of a sequencing batch reactor for aerobic oxidation of methane with denitrification (AeOM-D SBR). Dissolved $N_2O$ concentration increased, leveled-off and slightly decreased as the HRT increased from 0.25 to 1d. Concentration of the dissolved $N_2O$ was higher at the shorter HRT, which was highly associated with the lowered C/N ratio. A longer HRT resulted in a higher C/N ratio with a sufficient carbon source produced by methanotrophs via methane oxidation, which provided a favorable condition for reducing $N_2O$ formation. With a less formation of the dissolved $N_2O$, $N_2O$ emission rate was lower at a longer HRT condition due to the lower C/N ratio. Opposite to the $N_2O$ emission, emission rates of $CH_4$ and $CO_2$ were higher at a longer HRT. Longer HRT resulted in the greater total GHGs emission as $CO_2$ equivalent which was doubled when the HRT increased from 0.5d to 1.0 d. Contribution of $CH_4$ onto the total GHGs emission was most dominant accounting for 98 - 99% compared to that of $N_2O$ (< 2%).

• Proceedings of the Korea Water Resources Association Conference
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• 2006.05a
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• pp.1415-1419
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• 2006

메탄을 탄소원 및 에너지원으로 이용하는 메탄산화균은 물질대사과정 중에 다량의 세포외 고분자물질인 Extracellular polymeric substances(EPS)를 생성하는데, EPS는 카르복실기와 같은 표면흡착 기능을 가지고 있어 생체흡착제로 사용이 가능하다. 따라서 본 연구에서는 메탄산화세균을 이용하여 중금속인 Cd의 흡착성능을 파악하여 활성슬러지의 흡착능과 비교하고, EPS 농도별, pH별 흡착량의 변화를 실험한 후 Freundlich 흡착모델식에 적용하여 흡착공정의 기본적인 설계인자를 도출하고자 하였다. 실험에 사용한 메탄산화세균은 매립지 복토층 상부 토양에서 분리하여 실험실에서 대량으로 배양하였으며, EPS 생성을 위해 메탄을 Head space의 20%를 주입하고 $30^{\circ}C$, 150rpm에서 질소원이 부족한 조건으로 48hr 동안 배양하였다. Cd의 흡착실험은 용액의 pH를 3에서 8까지 변화를 주면서 활성슬러지와 메탄산화세균의 시간별 흡착능을 측정하였다. 또한 중금속의 농도별 흡착능을 측정하여 흡착평형 상수를 파악하였으며, 중금속 흡착 전, 후 미생물의 SEM 촬영, FT-IR 분석, 전자현미분석(EPMA)을 통하여 무기성분 분석 및 표면관찰을 수행하였다. 실험결과 메탄산화세균에 의해 생성된 EPS 물질은 중금속에 대한 강한 결합능력이 있으며, Cd에 대한 최고 흡착능은 26mg Cd(Ⅱ)/g VSS의 값을 보였다. 이러한 미생물의 EPS의 흡착능은 pH와 칼슘이온의 영향을 많이 받았으며, 메탄산화세균의 FT-IR 분석결과 EPS에는 sulfate ester, pyruvate 등과 같은 작용기와 amino sugar, carboxyl 작용기들이 많이 존재하여 활성슬러지에 비해 중금속의 흡착능이 높은 것으로 사료되었다.X>${\mu}_{max,A}$는 최대암모니아 섭취률을 이용하여 구한 결과 $0.65d^{-1}$로 나타났다.EX>$60%{\sim}87%$가 수심 10m 이내에 분포하였고, 녹조강과 남조강이 우점하는 하절기에는 5m 이내에 주로 분포하였다. 취수탑 지점의 수심이 연중 $25{\sim}35m$를 유지하는 H호의 경우 간헐식 폭기장치를 가동하는 기간은 물론 그 외 기간에도 취수구의 심도를 표층 10m 이하로 유지 할 경우 전체 조류 유입량을 60% 이상 저감할 수 있을 것으로 조사되었다.심볼 및 색채 디자인 등의 작업이 수반되어야 하며, 이들을 고려한 인터넷용 GIS기본도를 신규 제작한다. 상습침수지구와 관련된 각종 GIS데이타와 각 기관이 보유하고 있는 공공정보 가운데 공간정보와 연계되어야 하는 자료를 인터넷 GIS를 이용하여 효율적으로 관리하기 위해서는 단계별 구축전략이 필요하다. 따라서 본 논문에서는 인터넷 GIS를 이용하여 상습침수구역관련 정보를 검색, 처리 및 분석할 수 있는 상습침수 구역 종합정보화 시스템을 구축토록 하였다.N, 항목에서 보 상류가 높게 나타났으나, 철거되지 않은 검전보나 안양대교보에 비해 그 차이가 크지 않은 것으로 나타났다.의 기상변화가 자발성 기흉 발생에 영향을 미친다고 추론할 수 있었다. 향후 본 연구에서 추론된 기상변화와 기흉 발생과의 인과관계를 확인하고 좀 더 구체화하기 위한 연구가 필요할 것이다.게 이루어질 수 있을 것으로 기대된다.는 초과수익률이 상승하지만, 이후로는 감소하므로, 반전거래전략을 활용하는 경우 주식투자기간은 24개월이하의 중단기가 적합함을 발견하였다. 이상의 행태적 측면과 투자성과측면의 실증결과를 통하여 한국주식시장에 있어서 시장수익률을 평균적으로 초과할 수 있는 거래전

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.157-165
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• 2016

Under flooded rice fields, methanogens produce methane which comes out through rice stalks, thus rice fields are known as one of the anthropogenic sources of atmospheric methane. Studies have shown that use of manure increases amount of methane emission from rice. To investigate mechanisms by which manure boosts methane emission, comparative soil metagenomics between inorganically (NPK) and pig manure fertilized paddy soils (PIG) were conducted. Results from taxonomy analysis showed that more abundant methanogens, methanotrophs, methylotrophs, and acetogens were found in PIG than in NPK. In addition, BLAST results indicated more abundant carbohydrate mabolisetm functional genes in PIG. Among the methane metabolism related genes, PIG sample showed higher abundance of methyl-coenzyme M reductase (mcrB/mcrD/mcrG) and trimethylamine-corrinoid protein Co-methyltransferase (mttB) genes. In contrast, genes that down regulate methane emission, such as trimethylamine monooxygenase (tmm) and phosphoserine/homoserine phosphotransferase (thrH), were observed more in NPK sample. In addition, more methanotrophic genes (pmoB/amoB/mxaJ), were found more abundant in PIG sample. Identifying key genes related to methane emission and methane oxidation may provide fundamental information regarding to mechanisms by which use of manure boosts methane emission from rice. The study presented here characterized molecular variation in rice paddy, introduced by the use of pig manure.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.6
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• pp.991-1000
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• 2000

Methanotrophic consortium utilizing methane as the primary carbon source and secreting soluble methane monooxygenase (sMMO) was immobilized on celite R-635 to continuously treat a wastewater containing trichloroethylene (TCE). With influent 2 ppm of TCE. 80.4 and 84.5% of TCE was degraded in 6 and 20 hour of hydraulic retention time (HRT). respectively. and the removal efficiency of TCE was increased with an increase in HRT in methanotrophic consortium biofilm reactor (MCBR). With influent 5 ppm of TCE and 10 hour of HRT. average efficiency of TCE removal was decreased in initial stage. but gradually increased to 81%. TCE was degraded to 88.5 and 96.5% with 10 and 15 hour of HRT. respectively. when methane was supplied alternately with continuous oxygen supply at influent 5 ppm of TCE. The efficiency of TCE degradation was decreased probably because oxidation reaction of methane was proceeded slowly on MMO. when high concentration of methane was supplied with depletion of oxygen. As results of the pilot-scale study. biodegradation of TCE by MCBR system might be feasible at full-scale operation.