摘要
研究了大肠杆菌BL21(DE3)中重组胸苷磷酸化酶的最佳诱导表达条件及1-(2-脱氧-β-D-呋喃核糖基)-1,2,4-三唑-3-甲酰胺(1)的最佳生成条件。菌体培养3.5h时加入8mmol/L乳糖诱导6h后,胸苷磷酸化酶的酶活力为2.14×104u/mg湿菌体。5%湿菌体在pH7.0的0.1mol/L Tris-HCl缓冲液中与底物胸苷、1,2,4-三唑-3-甲酰胺于50℃反应5h后,所得1的转化率大于90%。
The induction and expression conditions of recombinant thymidine phosphorylase （Tpase） in E. coli BL （DE3） as well as the synthetic conditions of 1- （2-deoxy--β-D-ribofuranosyl） -1,2,4-triazole-3-carboxamide （1） were optimized. After the strain was cultured for 3.5h, 8mmol/L lactose was added and induced for 6h. The enzyme activity of Tpase reached to 2.14× 10^4u/mg wet cells. When 5 % wet cells were added into 0.1mol/L Tris-HC1 buffer solution （pH 7.0） and reacted for 5h at 50℃ with thymidine and 1,2,4-triazole-3-carboxyamide as the substrates, the conversion rate of 1 was over 90 %.
出处
《中国医药工业杂志》
CAS
CSCD
北大核心
2008年第8期584-587,共4页
Chinese Journal of Pharmaceuticals
基金
科技部科技型中小企业技术创新基金(07C26213100222)
