论文部分内容阅读
采用分子生物学方法构建16SrDNA基因文库,研究鸡粪沼气池发酵液中产甲烷菌的菌群结构。随机分析文库中50个克隆的16SrDNA基因序列,结果发现,其中46个克隆属于产甲烷菌属,与Methanogenium marinum strain AK-1菌株的相似性为99%-100%,占总数的92%;3个克隆(KD525、KD526和KD567)属于甲烷袋状菌属,与Methanoculleus sp.dm2菌株的相似性均为99%,占总数的6%;1个克隆(KD519)属于甲烷粒菌属,与Methanocorpusculum bavaricum菌株的相似性为99%,占总数的2%。另外,同样分析了样品中甲基辅酶M还原酶alpha亚基mcrA基因氨基酸序列的差异。气相色谱法分析结果显示,发酵液中甲酸含量为28.85g/L,约占总有机酸含量的81.7%;沼气的主要成分为甲烷、二氧化碳和氢气,分别约占总气体量的55.5%、41.1%和3.2%。
16SrDNA gene library was constructed by molecular biology method to study the flora structure of methanogen in the fermentation broth of chicken manure biogas. Randomly analyzing the 16SrDNA gene sequences of 50 clones in the library, 46 of them were identified as Methanogenas and 99% -100% similarity to Methanogenium marinum strain AK-1, accounting for 92% of the total. 3 The clones (KD525, KD526 and KD567) belong to the genus Methanococcus and all have 99% identity with Methanoculleus sp.dm2, accounting for 6% of the total. One clone (KD519) belongs to the genus Methanocorpusculum The similarity of bavaricum strains was 99%, accounting for 2% of the total. In addition, the amino acid sequences of the mcrA gene of methyl-Coenzyme M reductase alpha subunit in the sample were also analyzed. Gas chromatography analysis showed that the content of formic acid in fermentation broth was 28.85g / L, accounting for 81.7% of the total organic acid content. The main components of biogas were methane, carbon dioxide and hydrogen, accounting for 55.5% and 41.1 % And 3.2%.