酶促磷酸化法合成氚标记核苷三磷酸

SYNTHESIS OF TRITIUM LABELLED NUCLEOSIDE TRIPHOSPHATE BY ENZYMATIC PHOSPHORYLATION

摘要: 以 5 BrUMP、5 BrCMP、8 BrAMP及 8 BrGMP为原料,使用催化氚卤置换法制备了5-~3HUMP、5-~3HCMP、8-~3HAMP及8-~3HGMP。再用从啤酒鲜酵母中提取的粗酶,经酶促磷酸化,将5-~3HUMP、5-~3HCMP、8-~3HAMP及8-~3HGMP进一步合成了5-~3HUTP、5-~3HCTP、8-~3HATP及8-~3HGTP,并用纸层析方法进行纯化。作为副产品,还得到了氚标记核苷一磷酸和氚标记核苷二磷酸。产品的比活度为 14—19 Ci/mmol,放化纯度大于98％。

Abstract: (5-~3H)UMP, (5-~3H)CMP, (8-~3H)AMP and (8-~3H)GMP is prepared from 5Br-UMP, 5BrCMP, 8BrAMP and 8BrGMP by catalytic halogen--tritium replacement. (5-~3H)UTP, (5-~3H)CTP, (8-~3H)ATP and (8-~3H)GTP is subsequently synthe-sised from (5-~3H)UMP, (5-~3H)CMP, (8-~3H)AMP and (8-~3H)GMP by enzymaticphosphorylation with the crude enzyme preparaed from brewer's yeasts, andpurified by paper chromatography. In addition to tritium labelled nucleosidemonophosphate, diphosphate is obtained as a byproduct. The specific activities of these products are from 14 to 19 Ci/mmol and theirradiochemical purity is more than 98%.