Eguchi T, Kuge Y, Inoue K, Yoshikawa N, Mochida K, Uwajima T
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan.
Biosci Biotechnol Biochem. 1992 May;56(5):701-3. doi: 10.1271/bbb.56.701.
A new process for (6S)-tetrahydrofolate production from dihydrofolate was designed that used dihydrofolate reductase and an NADPH regeneration system. Glucose dehydrogenase from Gluconobacter scleroides KY3613 was used for recycling of the cofactor. The reaction mixture contained 200 mM dihydrofolate, 220 mM glucose, 2 mM NADP, 14.4 U/ml dihydrofolate reductase, and 14.4 U/ml Glucose dehydrogenase, and the reaction was complete after incubation at pH 8.0, and 40 degrees C for 2.5 hr. With (6S)-tetrahydrofolate as the starting material, l-leucovorin was synthesized via a methenyl derivative. The purity of the l-leucovorin was 100%, and its diastereomeric purity was greater than 99.5% d.e. as the (6S)-form.
设计了一种由二氢叶酸生产（6S）-四氢叶酸的新方法，该方法使用二氢叶酸还原酶和NADPH再生系统。来自硬脂葡萄糖杆菌KY3613的葡萄糖脱氢酶用于辅因子的循环利用。反应混合物包含200 mM二氢叶酸、220 mM葡萄糖、2 mM NADP、14.4 U/ml二氢叶酸还原酶和14.4 U/ml葡萄糖脱氢酶，在pH 8.0和40℃孵育2.5小时后反应完成。以（6S）-四氢叶酸为起始原料，通过甲烯基衍生物合成了L-亚叶酸。L-亚叶酸的纯度为100%，其非对映体纯度以（6S）-形式计大于99.5%de。
