ビタミンB_1関連化合物の代謝 : (VI)Saccharomyces cerevisiaeを用いるビタミンB_1構成のピリミジン体の定量法

概要

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Saccharomyces cerevisiae Hansen No. 0305 has an ability of synthesizing thiamine from 4-methyl-5-β-hydroxyethylthiazole (I) and 2-methyl-4-amino-5-hydroxymethylpyrimidine (II) or 2-methyl-4-amino-5-aminomethylpyrimidine (III). This thiamine-synthesizing activity was utilized for the determination of (II) or (III) in biological materials such as urine. Sacch. cerevisiae, preincubated in Reader's medium, was inoculated in the same medium containing the adequate amount of I and the sample ; then it was incubated at 37℃ for 40 hours. Thus synthesized thiamine was determined by the usual thiochrome method after boiling and hydrolysis by Taka-phosphatase. For the simplification and speed-up of the whole procedure, Sacch. cerevisiae was grown in the Reader's medium containing only I, at 25℃ overnight and then each 4ml portion of the whole medium was poured into a tube. It was incubated with an aliquot of the sample at 37℃ for 3 hours and the thiamine formed, could be estimated after boiling without Taka-phosphatase treatment. The dose-response curve of the sample was compared to the standard curve of (II) or (III) ; the value of (II) or (III) in the sample was computed from at least 2 corresponding points. The simplified procedure gave nearly the same results as the one incubating for 40 hours and the values found at the same sample by these procedures coincided fairly well on duplicated estimation.
日本ビタミン学会の論文
1961-09-25