Yuto YASHIRO
Isolation of novel methanogenic archaea by using co-culture method and specific cell sorting technique of flow-cytometry system.
Takashi YAMAGUCHI
It is difficult to selective cultivation of targeted methanogens from enrichment cultures by using the traditional cultivation isolation methods, because several methanogens having similar characteristics (i.e., substrate usage) often grow up in the enrichment cultures. Therefore, in this study, I developed a new isolation method of methanogens by use of specific cell sorting technique of flow-cytometry system. The experiments were carried out by use 11 methanogens that were selected by cover the phylogenetic classification, the cell morphology and the cell wall structure. As a result, it was possible that the isolation of targeted methanogen with 70-95% specificity from artificial mix culture of two types of methanogens. These results suggested that the new isolation method using flow-cytometry system was an effective method as isolation-supporting technique of methanogens.
In addition, I also attempted the isolation of taxonomically novel methanogens using co-culture method. Four methanogenic sludges were cultivated by the co-culture approach. After several attempets were made for over 1 year , I eventually succeeded in isolatation of a novel methanogen, belonging the group E1/E2 within the order Methanomicrobiales. After investigation of detailed characteristics of the methanogen, I justified the creation of a new species of a new genus for the methanogen, for which I proposed the name Methanoregula formicicum gen. nov., sp. nov
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