• Journal of Microbiology and Biotechnology
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• 제25권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.993-1003
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• 2013

Methanotrophs are the most important sink of $CH_4$, which is a more highly potent greenhouse gas than $CO_2$. Methanotrophic abundance and community diversity in cover soils from two typical semi-aerobic landfills (SALs) in China were detected using real-time polymerase chain reaction (real-time-PCR) and denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes, respectively. Real time-PCR showed that Type I methanotrophs ranged from $1.07{\times}10^6$ to $2.34{\times}10^7$ copies/g soil and that of Type II methanotrophs from $1.51{\times}10^7$ to $1.83{\times}10^8$ copies/g soil. The ratio of Type II to Type I methanotrophic copy numbers ranged from 5.61 to 21.89, indicating that Type II methanotrophs dominated in SAL. DGGE revealed that Type I methanotrophs responded more sensitively to the environment, changing as the community structure varied with different soil types and locations. Methylobacter, Methylosarcina, and Methylomicrobium for Type I, and Methylocystis for Type II were most prevalent in the SAL cover layer. Abundant interflow $O_2$ with high $CH_4$ concentration in SALs is the reason for the higher population density of methanotrophs and the higher enrichment of Type II methanotrophs compared with anaerobic landfills and other ecosystems, which proved a conclusion that increasing the oxygen supply in a landfill cover layer would greatly improve $CH_4$ mitigation.

• 대한환경공학회지
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• 제27권11호
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• pp.1198-1204
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• 2005

본 연구는 메탄산화균을 분리 배양하여 생물학적 질소 및 인 제거에 응용하는데 있다. 수도권 매립지의 상부 토양으로부터 NMS (nitrate mineral salt solution)배지로 분리 배양된 메탄산화균을 이용하여 영양염 제거 및 성장특성을 분석하였다. 분리 배양된 메탄산화균은 탈질의 탄소원으로서 이용될 수 있는 메탄올, 포름알데히드, 포름산으로 구성된 상당한 양의 유기물(COD 증가)을 생산하였다. 이때 메탄올의 생성속도는 $8\;mg/L{\cdot}hr$로 나타났다. 메탄산화균의 슬러지에 함유되어 있는 질소와 인의 함량을 볼 때 메탄산화균은 탈질에 필요한 탄소원 생성 뿐만 아니라 자체적으로 질소와 인을 성장기질로서 사용하는 것으로 나타났다.

• 한국미생물·생명공학회지
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• 제45권3호
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• pp.185-199
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• 2017

메탄은 자연적인 발생원과 인위적인 발생원에 의해 배출되며 지구온난화를 야기하는 대표적인 온실가스이다. 메탄을 탄소원과 에너지원으로 이용하는 메탄산화세균은 메탄의 생물학적 산화에 중요한 역할을 한다. 메탄산화세균의 서식지는 매우 다양하며 메탄산화반응의 핵심 효소인 methane monooxygenases (MMOs)는 메탄뿐 아니라 다른 기질을 산화할 수 있는 기질특이성을 가지고 있다. 이러한 메탄산화세균의 특성으로 인해 생물학적 메탄 저감 기술과 생물정화기술 분야에서 메탄산화세균의 활용에 대한 연구가 활발히 진행되고 있다. 본 총설 논문에서는 메탄산화세균의 종류, MMOs의 특성과 메탄산화세균의 고농도 배양 기술에 관한 최근 정보를 정리하였다. 또한 메탄산화세균을 이용한 생물학적 메탄 저감 관련 실험실 규모와 매립지 현장에서의 기술 개발 현황 및 적용 결과를 소개하였다. 이러한 생물학적 메탄 저감 시스템에서 메탄산화세균의 군집 거동 특성도 고찰하였다. 마지막으로, 메탄산화세균을 활용한 생물공학기술의 혁신을 위해 필요한 과제로 대사활성이 우수하거나 신규 대사능력을 가진 메탄산화세균의 지속적인 탐색 연구, 고농도 세포 대량배양기술 개발 및 미생물 컨소시움(메탄산화세균과 비메탄산화세균의 컨소시움) 디자인 및 관리 기술 등이 필요함을 제안하였다.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.360-370
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• 2012

Current ecological knowledge of methanotrophic biofilms is incomplete, although they have been broadly studied in biotechnological processes. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex 16S rDNA pyrosequencing was performed. A complete library (before being de-multiplexed) contained 33,639 sequences (average length, 415 nt). Interestingly, methanotrophs were not dominant, only making up 23% of the community. Methylosinus, Methylomonas, and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but less richer and diverse. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium, and Hyphomicrobium. The library was de-multiplexed into four libraries, with different sequencing efforts (3,915 - 20,133 sequences). Sorrenson abundance similarity results showed that the four libraries were almost identical (indices > 0.97), and phylogenetic comparisons using UniFrac test and P-test revealed the same results. It was demonstrated that the pyrosequencing was highly reproducible. These survey results can provide an insight into the management and/or manipulation of methanotrophic biofilms.

• 한국수처리학회지
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• 제26권6호
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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.

• Journal of Microbiology and Biotechnology
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• 제26권4호
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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.

• 한국미생물·생명공학회지
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• 제41권2호
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• pp.215-220
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• 2013

Tobermolite, perlite 및 polyurethane을 충전재로 이용한 바이오필터에 의한 메탄제거 기술이 개발되고 있다. 본 연구에서는 이들 충전재가 메탄산화능에 미치는 영향을 알아보았다. 습지토양과 매립지토양에서 분리한 혼합 메탄산화세균(consortium A, B, C and D)를 접종원으로 하고, 메탄산화속도와 메탄산화세균 수에 미치는 담체(perlite, tobermolite,polyurethane)의 영향을 조사하였다. Perlite를 첨가한 경우 메탄산화속도는 대조군 (담체 미첨가)보다 두 배 이상 증가하였고, 메탄산화세균 수도 10배 이상 증가하였다. Tobermolite를 첨가한 경우에는 일반세균 수 대비 메탄산화세균의 비율이 대조군과 다른 담체에 비해 높았다. 이는 tobermolite가 메탄산화세균이 우점할 수 있는 특이적 담체로 작용함을 시사한다. 이상의 결과로 부터 perlite와 tobermolite는 메탄산화세균의 활성을 증가시키는 서식지를 제공하며 메탄산화 공정시스템에 적용 시 좋은 담체의 역할을 할 수 있을 것으로 기대된다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.189.2-189
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• 2005

• Journal of Microbiology and Biotechnology
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• 제26권7호
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• pp.1234-1241
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• 2016

Methanol is a versatile compound that can be biologically synthesized from methane (CH₄) by methanotrophs using a low energy-consuming and environment-friendly process. Methylocella tundrae is a type II methanotroph that can utilize CH₄ as a carbon and energy source. Methanol is produced in the first step of the metabolic pathway of methanotrophs and is further oxidized into formaldehyde. Several parameters must be optimized to achieve high methanol production. In this study, we optimized the production conditions and process parameters for methanol production. The optimum incubation time, substrate, pH, agitation rate, temperature, phosphate buffer and sodium formate concentration, and cell concentration were determined to be 24 h, 50% CH₄, pH 7, 150 rpm, 30℃, 100 mM and 50 mM, and 18 mg/ml, respectively. The optimization of these parameters significantly improved methanol production from 0.66 to 5.18 mM. The use of alginate-encapsulated cells resulted in enhanced methanol production stability and reusability of cells after five cycles of reuse under batch culture conditions.