title: Enzymatic synthesis of 2'-deoxyadenosine and 6-methylpurine-2'-deoxyriboside by Escherichia coli DH5alpha overexpressing nucleoside phosphorylases from Escherichia coli BL21.
abstract: 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.
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Science, Fudan University, Shanghai 200433, PR China.
authors.forename: Shenghua
authors.initials: S
authors.lastname: Liang
authors.forename: Wenzhou
authors.initials: W
authors.lastname: Li
authors.forename: Tong
authors.initials: T
authors.lastname: Gao
authors.forename: Xiangying
authors.initials: X
authors.lastname: Zhu
authors.forename: Guimei
authors.initials: G
authors.lastname: Yang
authors.forename: Daming
authors.initials: D
authors.lastname: Ren
mesh_terms: 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
chemical_list: D003839:Deoxyadenosines; D011684:Purine Nucleosides; D011994:Recombinant Proteins; C032885:6-methylpurine 2'-deoxyriboside; D010430:Pentosyltransferases; C067995:nucleoside phosphorylase; C058118:2'-deoxyadenosine