Improved synthesis of 2'-deoxyadenosine and 5-methyluridine by Escherichia coli using an auto-induction system.
Nucleoside analogues are used widely for the treatment of viral diseases and cancer, however the preparation of some important intermediates of these nucleoside analogues, including 2'-deoxyadenosine (dAR) and 5-methyluridine (5-MU), remains inconvenient. To optimize the synthesis of dAR and 5-MU, recombinant strains and auto-induction medium were employed in this study. E. coli BL21(DE3) strains overexpressing purine nucleoside phosphorylase (PNP), uridine phosphorylase (UP) and thymidine phosphorylase (TP) were constructed and cultured in auto-induction ZYM-Fe-5052 medium for 8 h. The cultures of these strains were then used directly to synthesize dAR and 5-MU. Under optimized conditions, 30 mM adenine was converted to 29 mM dAR in 1 h, and 32 mM 5-MU was obtained from 60 mM thymine, using 6% (v/v) cell solutions as biocatalysts. These results indicate that our convenient and efficient method is ideal for the preparation of dAR and 5-MU, and has potential for the preparation of other nucleoside analogue intermediates.
School of Life Sciences, Tsinghua University, Beijing, 100084, China.
Juan
J
Xiong
Wenquan
W
Zhang
Jingtan
J
Su
Junlong
J
Shangguan
Yin
Y
Lin
Yang
Y
Yang
Rongqing
R
Zhang
Liping
L
Xie
Hongzhong
H
Wang
D003839:Deoxyadenosines / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D011683:Purine-Nucleoside Phosphorylase / Q000235:genetics / Q000378:metabolism; D013939:Thymidine Phosphorylase / Q000235:genetics / Q000378:metabolism; D014529:Uridine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D014543:Uridine Phosphorylase / Q000235:genetics / Q000378:metabolism
D003839:Deoxyadenosines; C009182:ribothymidine; D011683:Purine-Nucleoside Phosphorylase; D014543:Uridine Phosphorylase; D013939:Thymidine Phosphorylase; C058118:2'-deoxyadenosine; D014529:Uridine