利用嘧啶核苷磷酸化酶基因工程菌酶法合成5-氟尿苷的转化率影响因素的研究  

Study on Transformation of 5-FUR Produced by E.coli BL21(PyPNase)

作　　者：吴晓[1] 倪孟祥[1] 朱晓丽[1] 王旻[1] 吴梧桐[1] 

机构地区：[1]中国药科大学微生物制药教研室,江苏南京210009

出　　处：《药物生物技术》2008年第4期282-285,共4页Pharmaceutical Biotechnology

摘　　要：研究酶法合成5-氟尿苷(5-FUR)的反应条件。利用嘧啶核苷磷酸化酶(Pyrimidine-nucleosidephos-phorylase,PyPNase)基因工程菌大肠杆菌BL21(DE3)/pET28a+合成5-氟尿苷(5-FUR),考察不同反应条件对转化率的影响;尝试不同反应体系对转化率的影响。结果:(1)最佳转化条件:菌体:0.5WCG/L;UR:10mmol/L;5-FU:45mmol/L;PBS:40mmol/L;T:50℃;pH7.4;t:60min,转化率可达91.27%;(2)以Na2SO4溶液为溶剂,比以PBS溶液为溶剂的转化率略有提高。微生物酶法合成5-氟尿苷(5-FUR)的转化率较高,可考虑用于工业生产。To study the transformation condition of 5 FUR by E. coli BL21（PyPNase）. Using E. coli BL21（DE3）/pET28a＋ （Pyrimidine-nucleoside phosphorylase, PyPNase） to transform 5 FUR, the usual factors which affect the conversion rate and several kinds of solvent were studied. Results showed that （1）when the wet cell concentration of E. coli was 0. 5 WCG/L, the concentration of UR and 5-FUR was 10 mmol/L and 45 mmol/L respectively, the temperature and pH value were 50℃ and 7.4 respectively, the PBS concentration was 40 mmol/L, 91.27% UR was converted to FUR; （2） The conversion rate was higher when the Na2SO4 solution system was used. The conversion rate of 5-FUR is high enough to be used in commercial production.

关 键 词：基因工程菌BL21 嘧啶核苷磷酸化酶 5-氟尿苷 微生物转化 转化率 

分 类 号：Q78[生物学—分子生物学]