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

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

• BIOTECHNOLOGY AND BIOENGINEERING • Previous Articles Next Articles

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. 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)

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. 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

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)

Abstract

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

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

DING Qingbao, OU Ling, WEI Dongzhi, WEI Xiaokun, XU Yanmei, ZHANG Chunyan. Induction of Recombinant Uridine Phosphorylase and Its Application in Biosynthesis of Pyrimidine Nucleosides[J]. , 2011, 19(1): 122-127.

丁庆豹, 欧伶, 魏东芝, 魏晓琨, 许彦梅, 张春艳. Induction of Recombinant Uridine Phosphorylase and Its Application in Biosynthesis of Pyrimidine Nucleosides[J]. , 2011, 19(1): 122-127.

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

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

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