Enzymatic synthesis of 2'-deoxyadenosine and 6-methylpurine-2'-deoxyriboside by Escherichia coli DH5alpha overexpressing nucleoside phosphorylases from Escherichia coli BL21.
Genes encoding purine nucleoside phosphorylase (deo D), uridine phosphorylase (udp) and thimidine phosphorylase (deo A) from Escherichia coli BL21 were cloned and overexpressed in E. coli DH5alpha. The recombinant strains were employed to synthesize 2'-deoxyadenosine (dAR) and 6-methylpurine-2'-deoxyriboside (MePdR). Experimental parameters such as strains, temperature, pH, reagent concentration and cell mass were optimized. Under the optimal situation, 96% adenine was converted to dAR and 95% 6-methylpurine (MeP) was converted to MePdR in an hour, using 0.2 per thousand (dry wt./v) cell paste as biocatalyst.
Liang
Shenghua
S
State Key Laboratory of Genetic Engineering, School of Life Science, Fudan University, Shanghai 200433, PR China.
Li
Wenzhou
W
Gao
Tong
T
Zhu
Xiangying
X
Yang
Guimei
G
Ren
Daming
D
D003839:Deoxyadenosines / Q000096:biosynthesis*; D004926:Escherichia coli / Q000502:physiology*; D010430:Pentosyltransferases / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011684:Purine Nucleosides / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000378:metabolism
D003839:Deoxyadenosines; D011684:Purine Nucleosides; D011994:Recombinant Proteins; C032885:6-methylpurine 2'-deoxyriboside; D010430:Pentosyltransferases; C067995:nucleoside phosphorylase; C058118:2'-deoxyadenosine