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

被引:4
作者
Ding Qingbao [1 ]
Ou Ling [1 ]
We Dongzhi [1 ]
We Xiaokun [2 ]
Xu Yanmei [2 ]
Zhang Chunyan [2 ]
机构
[1] E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China
[2] Shanghai Pucheng Biosci & Biotechnol Co Ltd, Shanghai 200235, Peoples R China
关键词
5-fluorouridine; lactose; uridine phosphorylase; pyrimidine nucleoside; uridine; 5-fluorouracil; ENZYMATIC-SYNTHESIS; BACILLUS-STEAROTHERMOPHILUS; ENTEROBACTER-AEROGENES; ADENINE-ARABINOSIDE; PURIFICATION; EXPRESSION;
D O I
10.1016/S1004-9541(09)60187-7
中图分类号
TQ [化学工业];
学科分类号
0817 ;
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 degrees C. Wet cells of pUDP was used as catalyst to biosynthesize 5-fluorouridine from 30 mmol.L-1 uridine and 5-fluorouracil in phosphate buffer (pH 7.0) catalyzed at 50 degrees C 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%.
引用
收藏
页码:122 / 127
页数:6
相关论文
共 16 条
[1]  
[Anonymous], 1989, MOL CLONING LAB MANU
[2]  
BENJAMIN CR, 1973, J BIOL CHEM, V248, P2040
[3]   Antiviral drugs: current state of the art [J].
De Clercq, E .
JOURNAL OF CLINICAL VIROLOGY, 2001, 22 (01) :73-89
[4]   Review: Optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter [J].
Donovan, RS ;
Robinson, CW ;
Glick, BR .
JOURNAL OF INDUSTRIAL MICROBIOLOGY, 1996, 16 (03) :145-154
[5]   Nucleoside analogues and nucleobases in cancer treatment [J].
Galmarini, CM ;
Mackey, JR ;
Dumontet, C .
LANCET ONCOLOGY, 2002, 3 (07) :415-424
[6]   THE ENZYMATIC-SYNTHESIS OF ANTIVIRAL AGENTS [J].
HANRAHAN, JR ;
HUTCHINSON, DW .
JOURNAL OF BIOTECHNOLOGY, 1992, 23 (02) :193-210
[7]   SYNTHESIS OF 5-METHYLURIDINE BY A THERMOPHILE, BACILLUS-STEAROTHERMOPHILUS JTS859 [J].
HORI, N ;
WATANABE, M ;
YAMAZAKI, Y ;
MIKAMI, Y .
AGRICULTURAL AND BIOLOGICAL CHEMISTRY, 1989, 53 (01) :197-202
[8]  
ISHII N, 1989, AGR BIOL CHEM TOKYO, V52, P3209
[9]   PURIFICATION AND CHARACTERIZATION OF A NOVEL NUCLEOSIDE PHOSPHORYLASE FROM A KLEBSIELLA SP AND ITS USE IN THE ENZYMATIC PRODUCTION OF ADENINE-ARABINOSIDE [J].
LING, F ;
INOUE, Y ;
KIMURA, A .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, 56 (12) :3830-3834
[10]   Use of lactose to induce expression of soluble NifA protein domains of Herbaspirillum seropedicae in Escherichia coli [J].
Monteiro, RA ;
Souza, EM ;
Yates, MG ;
Pedrosa, FO ;
Chubatsu, LS .
CANADIAN JOURNAL OF MICROBIOLOGY, 2000, 46 (11) :1087-1090