• 제목/요약/키워드: Methanosarcina mazei

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Isolation and characterization of new Methanosarcina mazei strains KOR-3, -4, -5, and -6 from an anaerobic digester using pig slurry

  • Battumur, Urantulkhuur;Yoon, Youngman;Bae, Gui Sek;Kim, Chang-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권8호
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    • pp.1198-1205
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    • 2017
  • Objective: An experiment was conducted to isolate and identify new methanogens in Korea from an anaerobic digester that uses pig slurry. Methods: An anaerobic digestate sample was collected from an anaerobic digester using pig slurry. Pre-reduced media were used for the growth and isolation of methanogens. Growth temperature range, pH range, NaCl concentration range, substrate utilization, and antibiotic tolerance were investigated to determine the physiological characteristics of isolated methanogens. The isolates were also examined microscopically for their morphology and Gram-stained. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification purpose. Results: Four strains, designated KOR-3, -4, -5, and -6, were isolated and were non-motile, irregular coccoid, and 0.5 to $1.5{\mu}m$ in diameter. Moreover, the cell walls of isolated strains were Gram-negative. KOR-3 and KOR-4 strains used acetate for methane production but did not use $H_2+CO_2$, formate, or methanol as a growth substrate KOR-5 and KOR-6 strains utilized acetate, methanol, and trimethylamine for methanogenesis but did not use $H_2+CO_2$ or formate as a growth substrate. The optimum temperature and pH for growth of four strains were $39^{\circ}C$ and 6.8 to 7.2, respectively. The optimum concentration of NaCl for growth of KOR-3, KOR-5, and KOR-6 were 1.0% (w/v). The optimum NaCl concentration for KOR-4 was 0.5% (w/v). All of the strains tolerated ampicillin, penicillin G, kanamycin, streptomycin, and tetracycline; however, chloramphenicol inhibited cell growth. Phylogenetic analysis of 16S rRNA and mcrA genes demonstrated that strains KOR-3, -4, -5, and -6 are related to Methanosarcina mazei (M. mazei, 99% sequence similarity). Conclusion: On the basis of physiological and phylogenetic characteristics, strains KOR-3, -4, -5, and -6 are proposed to be new strains within the genus Methanosarcina, named M. mazei KOR-3, -4, -5, and -6.

농축산(農畜産) 폐기물(廢棄物) 처리(處理)를 위(爲)한 저온내성(低溫耐性) 메탄 생성균(生成菌)의 특성(特性)에 관(關)한 연구(硏究);III. 저온내성(低溫耐性) Methanogens의 분리(分離) (Study on Low Temperature Tolerant Methane-Producing Bacteria for the Treatment of Agricultural and Livestock Wastes;III. Isolation of Low Temperature Tolerant Methanogens)

  • 정광용;김재정
    • 한국환경농학회지
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    • 제15권3호
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    • pp.362-371
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    • 1996
  • 농축산(農畜産) 폐기물(廢棄物)의 혐기적(嫌氣的) 처리공정(處理工程)에 적용하기 위하여 아한대(亞寒帶) 지역으로부터 분리(分離)한 Methanogen의 생화학적(生化學的) 특성(特性)을 조사한 결과는 다음과 같다. 저온(低溫)에서도 메탄생성량(生成量)이 많았던 늪지 퇴적물(堆積物)(Canada, $56.9^{\circ}$ N)에서 분리(分離)된 균(菌)은 Methanobacterium M-251, Methanobacterium M-253, 호수(湖水) 퇴적물(堆積物) IV(Canada, $55.0^{\circ}$ N)에서 분리(分離)된 균(菌)은 Methanosarcina mazei M-372, 갯벌흙 II(Korea, $37.0^{\circ}$ N)에서 분리(分離)된 균(菌)은 Methanobacterium formicicum M-375로 각각 동정되었다. 분리(分離)된 4종의 메탄균은 당분해능은 없으며 적정 pH는 M-251, M-375는 6.8, M-253은 6.5, 그리고 M-372는 7.0이었다. 최적(最適) 생육온도(生育溫度)는 M-251과 M-253은 $30^{\circ}C$이었고 M-272와 M-375는 $37^{\circ}C$이었다. 최저(最低) 생육온도(生育溫度)는 M-251, M-253은 $8^{\circ}C$이었고, M-372, M-375는 $13^{\circ}C$이었다. 분리균주(分離菌株)의 생육량(生育量)은 $30^{\circ}C$에 비하여 $17.5^{\circ}C$에서 배양(培養)할 때 $32{\sim}50%$ 정도 낮았다. M-251, M-253, M-375의 균체(菌體) 생산량(生産量)은 $0.38{\sim}1.21$ g/1M $CH_4$ 범위 이었으며 M-372는 $1.14{\sim}1.51$ $g/CH_4$범위이었다.

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