Shibatani T, Nishimura N, Nabe K, Kakimoto T, Chibata I
Appl Microbiol. 1974 Apr;27(4):688-94. doi: 10.1128/am.27.4.688-694.1974.
To develop an efficient method for the production of urocanic acid, optimal conditions for the production of microbial L-histidine ammonia lyase and for the conversion of L-histidine to urocanic acid by this enzyme were studied. A number of microorganisms were screened to test their ability to form and accumulate urocanic acid from L-histidine. Achromobacter liquidum was selected as the best organism. With this organism, enzyme activity as high as 2.0 units/ml could be produced by a shaking culture at 30 C in a medium containing glucose, urea, potassium phosphate, L-histidine, yeast extract, peptone, and inorganic salts. Appropriate addition of a surface-active agent to the reaction mixture shortened the time required for the conversion. A large amount of L-histidine was converted stoichiometrically to urocanic acid in 48 h at 40 C. Accumulated urocanic acid was readily isolated in pure form by ordinary procedures with isoelectric precipitation. Yields of isolated urocanic acid of over 92% from L-histidine were easily attainable. When the culture of Achromobacter liquidum was added to DL-histidine, D-histidine and urocanic acid were simultaneously obtained in high yields.
为开发一种高效生产尿刊酸的方法,研究了微生物L-组氨酸解氨酶生产的最佳条件以及该酶将L-组氨酸转化为尿刊酸的最佳条件。筛选了多种微生物以测试它们从L-组氨酸形成并积累尿刊酸的能力。选择了液化无色杆菌作为最佳菌株。使用该菌株,在含有葡萄糖、尿素、磷酸钾、L-组氨酸、酵母提取物、蛋白胨和无机盐的培养基中于30℃振荡培养,可产生高达2.0单位/毫升的酶活性。向反应混合物中适当添加表面活性剂可缩短转化所需时间。在40℃下,大量的L-组氨酸在48小时内按化学计量转化为尿刊酸。通过等电沉淀的常规方法可轻松分离出积累的纯尿刊酸。从L-组氨酸中分离得到的尿刊酸产率很容易超过92%。当将液化无色杆菌培养物添加到DL-组氨酸中时,可同时高产获得D-组氨酸和尿刊酸。