Acclimated H
2/CO
2 using methanogen was cultured using stopped tube, with the production of vitamin B
12 as a by-product. And the process of extraction and refining of vitamin B
12 from culture liquid were studied. The XAD-2 resin was used for separation of vitamin B
12, and more than 90% of vitamin B
12 yield was achieved by modification for vitamin B
12 extraction and refining method used in this study. Under light and shade-culture condition, it was indicated that shading condition was required when the goal of methanogen cultivation is to produce vitamin B
12. Furthermore, in order to extract vitamin B
12 suitably from intracellular, the process of freezing and thawing for methanogen is necessary. Also, the highest vitamin B
12 concentration was obtained when methanogen having a high activity in logarithmic growth phase. Therefore, it was found that, the logarithmic growth stage was important for vitamin B
12 production from methane fermentation.
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