Thermophilic phosphoribosyltransferases HB27 in nucleotide synthesis.

作者信息

Fateev Ilja V, Sinitsina Ekaterina V, Bikanasova Aiguzel U, Kostromina Maria A, Tuzova Elena S, Esipova Larisa V, Muravyova Tatiana I, Kayushin Alexei L, Konstantinova Irina D, Esipov Roman S

机构信息

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya Str., 16/10, Moscow, GSP-7, 117997, Russia.

出版信息

Beilstein J Org Chem. 2018 Dec 21;14:3098-3105. doi: 10.3762/bjoc.14.289. eCollection 2018.

Phosphoribosyltransferases are the tools that allow the synthesis of nucleotide analogues using multi-enzymatic cascades. The recombinant adenine phosphoribosyltransferase (APRT) and hypoxanthine phosphoribosyltransferase (HPRT) from HB27 were expressed in strains and purified by chromatographic methods with yields of 10-13 mg per liter of culture. The activity dependence of APRT and HPRT on different factors was investigated along with the substrate specificity towards different heterocyclic bases. The kinetic parameters for HPRT with natural substrates were determined. Two nucleotides were synthesized: 9-(β-D-ribofuranosyl)-2-chloroadenine 5'-monophosphate (2-Сl-AMP) using APRT and 1-(β-D-ribofuranosyl)pyrazolo[3,4-]pyrimidine-4-one 5'-monophosphate (Allop-MP) using НPRT.