Enzymatic synthesis of adenosine-5'-phosphate from inosine-5'-phosphate.
Enzymatic synthesis of adenosine-5'-phosphate from inosine-5'-phosphate.
复制标题
从 5-磷酸肌苷酶促合成 5-磷酸腺苷。
DOI：
10.1016/s0021-9258(18)65142-7
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发表时间：
1956
期刊：
影响因子：
--
通讯作者：
I. Lieberman
中科院分区：
文献类型：
--
作者：
I. Lieberman
材料与方法无细胞提取物-E的制备。大肠杆菌，菌株 B，在葡萄糖和无机盐培养基2上以 37” 剧烈摇动生长。当用科尔曼初级分光光度计在 540 rnp 测量的光密度约为 0.7 时，冷却培养物，并通过在 3” 下离心收集细胞。通过用氧化铝（Alcoa A-301，每克包装的湿细胞 2.5 克）研磨（6）并用磷酸钾缓冲液（0.005 M，pH 7.2，每克湿细胞 5.5 毫升）提取来制备无细胞提取物。通过离心（约10,000 X g）除去不溶性物质。Materials-15P和ATP以结晶钠盐形式从Sigma Chemical Company获得。 GDP、GTP、UTP、IDP 和 ITP 均来自同一来源，并通过阴离子交换或纸层析进一步纯化。 G5P、ADP（Sigma Chemical Company）和胞苷二磷酸和三磷酸（Pabst Brewing Company）无需进一步纯化即可使用。
Materials and MethodsPreparation of Cell-Free Extract-E. coli, strain B, was grown on a glucose and inorganic salts medium2 at 37” with vigorous shaking. When the optical density, measured with a Coleman junior spectrophotometer at 540 rnp, was approximately 0.7, the cultures were cooled, and the cells were collected by centrifugation at 3”. Cell-free extracts were prepared by grinding with alumina (Alcoa A-301, 2.5 gm. per gm. of packed, wet cells)(6) and extracting with potassium phosphate buffer (0.005 M, pH 7.2, 5.5 ml. per gm. of wet cells). Insoluble material was removed by centrifugation (approximately 10,000 X g).Materials-15P and ATP were obtained from the Sigma Chemical Company as the crystalline sodium salts. GDP, GTP, UTP, IDP, and ITP were obtained from the same source and were further purified by anion exchange or paper chromatography. G5P, ADP (Sigma Chemical Company), and cytidine di-and triphosphate (Pabst Brewing Company) were used without further purification.