Study on Low Temperature Tolerant Methane-Producing Bacteria for the Treatment of Agricultural and Livestock Wastes;III. Isolation of Low Temperature Tolerant Methanogens

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)

This study was conducted to investigate the biochemical properties of isolated bacteria, low temperature tolerant methanogens which were selected for use as inoculum for anaerobic fermentation of agricultural and livestock wasted at low temperature. The results, obtained were summarized as follows: Low temperature tolerant methanogens were isolated from the samples which showed the high methanogenesis rate by enrichment culture at low temperature in methanol medium. These methanogens, Methanobacterium M-251 and Methanobacterium M-253 were isolated from swampy sediment at latitude $56.9^{\circ}$, Methanosarcina mazei M-372 from lake sediment IV at latitude $55.0^{\circ}$ N, and Methanobacterium formicicum M-375 from tidal land soil at latitude $37.0^{\circ}N$, respectively. The isolated anaerobic bacteria could not use sugars as carbon sources. The optimum pH value for the growth of M-251 and M-375 was 6.8, but those for M-253 and M-372 6.5 and 7.0, respectively. The minimum growth temperature of isolated, M-251 and M-253 were $8^{\circ}C$ and the optimum temperature $30^{\circ}C$, while the minimum of M-392 and M-395 were $13^{\circ}C$ and the optimum $37^{\circ}C$. The growth rate of isolates at $17.5^{\circ}C$ were lower by 32-50% than that of $30^{\circ}C$. The isolated Methanobacterium strains such as M-251, M-253, and M-375 have lower cell yield, 0.38-1.21g/1M $CH_4$ than 1.14-1.51g/1M $CH_4$ of Methanosarcina mazei M-372.

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.