CIESC Journal ›› 2011, Vol. 19 ›› Issue (1): 122-127.

• • 上一篇    下一篇

Induction of Recombinant Uridine Phosphorylase and Its Application in Biosynthesis of Pyrimidine Nucleosides

丁庆豹1, 欧伶1, 魏东芝1, 魏晓琨2, 许彦梅2, 张春艳2   

  1. 1. State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;
    2. Shanghai Pucheng Bioscience and Biotechnology Co., Ltd., Shanghai 200235, China
  • 收稿日期:2010-02-01 修回日期:2010-11-27 出版日期:2011-02-28 发布日期:2011-02-28
  • 通讯作者: WEI Dongzhi, E-mail: dzhwei@ecust.edu.cn
  • 基金资助:
    Supported by“Production,Education&Research”item of Shanghai Baoshan(08-H-4)

Induction of Recombinant Uridine Phosphorylase and Its Application in Biosynthesis of Pyrimidine Nucleosides

DING Qingbao1, OU Ling1, WEI Dongzhi1, WEI Xiaokun2, XU Yanmei2, ZHANG Chunyan2   

  1. 1. State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;
    2. Shanghai Pucheng Bioscience and Biotechnology Co., Ltd., Shanghai 200235, China
  • Received:2010-02-01 Revised:2010-11-27 Online:2011-02-28 Published:2011-02-28
  • Supported by:
    Supported by“Production,Education&Research”item of Shanghai Baoshan(08-H-4)

摘要: Recombinant Escherichia coli pUDP,which overexpressed uridine phosphorylase(UPase),was constructed.0.5 mmol·L-1 lactose had a similar induction effect as the commonly used inducer IPTG during 2.5-5.5 h of cell growth.The lactose-induced UPase was stable at 50℃.Wet cells of pUDP was used as catalyst to biosynthesize 5-fluorouridine from30 mmol·L-1 uridine and 5-fluorouracil in phosphate buffer(pH 7.0)catalyzed at 50℃ for 1.5 h and the yield of 5-fluorouridine was higher than 68%.Under the optimum reaction conditions for production of 5-fluorouridine,5-methyluridine and azauridine were synthesized from uridine by pUDP,the yield was 61.7% and 47.2% respectively.Deoxynucleosides were also synthesized by pUDP,but the yield was only about 20%.

关键词: 5-fluorouridine, lactose, uridine phosphorylase, pyrimidine nucleoside, uridine, 5-fluorouracil

Abstract: Recombinant Escherichia coli pUDP,which overexpressed uridine phosphorylase(UPase),was constructed.0.5 mmol·L-1 lactose had a similar induction effect as the commonly used inducer IPTG during 2.5-5.5 h of cell growth.The lactose-induced UPase was stable at 50℃.Wet cells of pUDP was used as catalyst to biosynthesize 5-fluorouridine from30 mmol·L-1 uridine and 5-fluorouracil in phosphate buffer(pH 7.0)catalyzed at 50℃ for 1.5 h and the yield of 5-fluorouridine was higher than 68%.Under the optimum reaction conditions for production of 5-fluorouridine,5-methyluridine and azauridine were synthesized from uridine by pUDP,the yield was 61.7% and 47.2% respectively.Deoxynucleosides were also synthesized by pUDP,but the yield was only about 20%.

Key words: 5-fluorouridine, lactose, uridine phosphorylase, pyrimidine nucleoside, uridine, 5-fluorouracil