[Synthesis of 2-chloro-2'-deoxyadenosine by microbiological transglycosylation using a recombinant Escherichia coli strain].
Cladribine (2-chloro-2'-deoxyadenosine) was synthesized using intact cells of the recombinant Escherichia coli strain producing Geobacillus stearothermophilus B-2194 thermostable purine-nucleoside phosphorylase II (EC 2.4.2.1). Use of the cells containing this thermostable enzyme allowed the process to be conducted at a temperature of 70 degrees C, which provided the maximal concentrations of sparingly soluble substrates. The best results were obtained with 2-chloroadenine as a modified base. The highest yield of the target 2-chloro-2'-deoxyadenosine (up to 95% in the case of deoxyguanosine) was reached when using 2'-deoxypurines as donors of deoxyribose. Use of thymidine for these purposes required its considerable molar excess over 2-chloroadenine (up to 6 : 1), which is connected with a nonoptimal amount of endogenous thymidine phosphorylase, necessary for synthesis of deoxyribose-1-phosphate, in the transglycosylation reaction.
Taran
S A
SA
Verevkina
K N
KN
Esikova
T Z
TZ
Feofanov
S A
SA
Miroshnikov
A I
AI
D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism; D017338:Cladribine / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D006031:Glycosylation; D011683:Purine-Nucleoside Phosphorylase / Q000378:metabolism*; D013696:Temperature; D013936:Thymidine / Q000378:metabolism; D013939:Thymidine Phosphorylase / Q000378:metabolism
C069484:2-chloroadenine; D017338:Cladribine; D011683:Purine-Nucleoside Phosphorylase; D013939:Thymidine Phosphorylase; D000225:Adenine; D013936:Thymidine