JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2010 Vol.11 No.11 P.880-888

Enzymatic synthesis of nucleosides by nucleoside phosphorylase co-expressed in Escherichia coli

Author(s): Qing-bao Ding, Ling Ou, Dong-zhi Wei, Xiao-kun Wei, Yan-mei Xu, Chun-yan Zhang
Affiliation(s): State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China, Shanghai Scibest Biotechnology Co., Ltd., Shanghai 200235, China
Corresponding email(s): bioxianleid@163.com, dzwei@ecust.edu.cn
Key Words: Nucleoside phosphorylase, Lactose, Enzymatic synthesis

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Abstract: nucleoside phosphorylase is an important enzyme involved in the biosynthesis of nucleosides. In this study, purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase were co-expressed in Escherichia coli and the intact cells were used as a catalyst for the biosynthesis of nucleosides. For protein induction, lactose was used in place of isopropyl β-D-1-thiogalactopyranoside (IPTG). When the concentration of lactose was above 0.5 mmol/L, the ability to induce protein expression was similar to that of IPTG. We determined that the reaction conditions of four bacterial strains co-expressing these genes (TUD, TAD, DUD, and DAD) were similar for the biosyntheses of 2,6-diaminopurine nucleoside and 2,6-diaminopurine deoxynucleoside. When the substrate concentration was 30 mmol/L and 0.5% of the recombinant bacterial cell volume was used as the catalyst (pH 7.5), a greater than 90% conversion yield was reached after a 2-h incubation at 50 °C. In addition, several other nucleosides and nucleoside derivatives were efficiently synthesized using bacterial strains co-expressing these recombinant enzymes.

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