An efficient synthesis of polyguanylic acid by a thermophilic polynucleotide phosphorylase

Yo Kikuchi, Keiko Hirai, Fumio Hishinuma, Kenji Sakaguchi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Polyguanylic acid (poly(G)) was synthesized from GDP in a yield of 60-75% by Thermus thermophilus polynucleotide phosphorylase (polyribonucleotide: orthophosphate nucleotidyltransferase, EC 2.7.7.8) at 70°C, pH 8.5 in the presence of Mg2+. The yield was dependent on the ratio of GDP to Mg2+, but was independent of the concentrations of enzyme or substrate. The maximal rate of GDP polymerization was obtained when the ratio of GDP to Mg2+ was 3 : 1. However, by prolonged incubation, the higher initial ratio of over 4 : 1 was preferred because of the rapid consumption of GDP in the reaction mixture. Poly(G) prepared by 1 h incubation was heterogeneous in size from 5 S to over 23 S, but by prolonged incubation of 19 h the size of product converged to 9 S as judged by sucrose density gradient centrifugation. Its chain length was determined by terminal nucleoside analysis to be 200 nucleotides long.

Original languageEnglish
Pages (from-to)287-294
Number of pages8
JournalBBA Section Nucleic Acids And Protein Synthesis
Volume476
Issue number4
DOIs
Publication statusPublished - 1977 Jun 17
Externally publishedYes

ASJC Scopus subject areas

  • Medicine(all)

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