Mitsuda H, Takii Y, Iwami K, Yasumoto K
J Nutr Sci Vitaminol (Tokyo). 1975;21(1):19-26. doi: 10.3177/jnsv.21.19.
Evidence was presented by paper chromatographic analysis on the occurrence of an enzyme capable of catalyzing a pyrophosphate transfer from ATP to thiamine in green leaves of various plants. The exclusive localization of the enzyme activity in the 105,000 X g supernatant (in a soluble form) was demonstrated by differential centrifugation of a cell homogentae in 0.25 M sucrose. The enzyme was purified by column chromatography with DEAE-cellulose and by gel filtration with Sephadex G-150. The partially pruified preparation, while contaminated with detectable activity of acid phosphatase, lost the ability of utilizing thiamine monophosphate as the substrate in place of thiamine. These findings lead to the conclusion that thiamine pyrophosphate is formed in green leaves of plants through a direct pyrophosphorylation of thiamine in the presence of ATP and Mg.
通过纸色谱分析提供了证据，证明在各种植物的绿叶中存在一种能够催化焦磷酸从ATP转移到硫胺素的酶。通过在0.25M蔗糖中对细胞匀浆进行差速离心，证明了酶活性仅定位在105,000 X g上清液中（以可溶形式存在）。该酶通过用DEAE - 纤维素进行柱色谱和用Sephadex G - 150进行凝胶过滤进行纯化。部分纯化的制剂虽然被检测到的酸性磷酸酶活性污染，但失去了利用硫胺素单磷酸代替硫胺素作为底物的能力。这些发现得出结论，植物绿叶中的硫胺素焦磷酸是在ATP和镁存在下通过硫胺素的直接焦磷酸化形成的。
