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On-line Access: 2010-11-04

Received: 2010-05-30

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Crosschecked: 2010-10-11

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

http://doi.org/10.1631/jzus.B1000193


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


Qing-bao Ding, Ling Ou, Dong-zhi Wei, Xiao-kun Wei, Yan-mei Xu, Chun-yan Zhang. Enzymatic synthesis of nucleosides by nucleoside phosphorylase co-expressed in Escherichia coli[J]. Journal of Zhejiang University Science B, 2010, 11(11): 880-888.

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author="Qing-bao Ding, Ling Ou, Dong-zhi Wei, Xiao-kun Wei, Yan-mei Xu, Chun-yan Zhang",
journal="Journal of Zhejiang University Science B",
volume="11",
number="11",
pages="880-888",
year="2010",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000193"
}

%0 Journal Article
%T Enzymatic synthesis of nucleosides by nucleoside phosphorylase co-expressed in Escherichia coli
%A Qing-bao Ding
%A Ling Ou
%A Dong-zhi Wei
%A Xiao-kun Wei
%A Yan-mei Xu
%A Chun-yan Zhang
%J Journal of Zhejiang University SCIENCE B
%V 11
%N 11
%P 880-888
%@ 1673-1581
%D 2010
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000193

TY - JOUR
T1 - Enzymatic synthesis of nucleosides by nucleoside phosphorylase co-expressed in Escherichia coli
A1 - Qing-bao Ding
A1 - Ling Ou
A1 - Dong-zhi Wei
A1 - Xiao-kun Wei
A1 - Yan-mei Xu
A1 - Chun-yan Zhang
J0 - Journal of Zhejiang University Science B
VL - 11
IS - 11
SP - 880
EP - 888
%@ 1673-1581
Y1 - 2010
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000193


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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