采用产氨短杆菌合成辅酶A的研究

STUDIES ON CoA SYNTHESIZED BY BREVIBACTERIUM AMMONIAGENES

用产氨短杆菌鲜菌体在以腺嘌呤代替ATP的反应系统中酶促合成辅酶A(CoA),每毫升反应液的 CoA 产量达221单位(u);在不加任何核苷酸类物质的条件下,也可达185u;合成在5小时左右达高峰。CoA 合成主要在细胞内进行。提出了连续式碳-阴离子交换树脂柱提纯CoA 的新方法,既提高了收率,又避免了原锌汞齐法的毒物危害,方法简易可行。用本法制得的结晶品含 CoA 231u/mg,收率17.0%;阴柱洗脱液不经结晶直接制成 CoA 注射液,提取率20.9%,经广州药检所检验,各项指标均符合国家标准。

CoA was enzymatically synthesized by Brevibacterium ammoniagenes,using adenine inplace of ATP as a precursor.The amount of synthesizing CoA was 221 units in every milliterfor the reaction mixture.If no adenine was used in the CoA synthesis reaction system,theamount of synthesizing CoA was as high as 185 units per milliliter.The optimum time of syn-thesizing CoA was about 5h.CoA was mainly formed inside the bacterial cell.We had pro-vided a new process of purifying CoA by a series of columns involving carbon and anion ex-change resin.The process not only increased the yield of CoA,but also avoided the poisonfrom applying the Zn-Hg amalgamation process.It also simplified the operation.Crystal CoA obtained by this new method contained CoA 231 units per milligram,andthe yield was 17.0 percent.If final CoA elution was directly collected,the yield could be rai-sed to 20.9 percent.