TY - JOUR
T1 - An efficient synthesis of polyguanylic acid by a thermophilic polynucleotide phosphorylase
AU - Kikuchi, Yo
AU - Hirai, Keiko
AU - Hishinuma, Fumio
AU - Sakaguchi, Kenji
PY - 1977/6/17
Y1 - 1977/6/17
N2 - 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.
AB - 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.
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U2 - 10.1016/0005-2787(77)90293-3
DO - 10.1016/0005-2787(77)90293-3
M3 - Article
C2 - 884104
AN - SCOPUS:0017412790
VL - 476
SP - 287
EP - 294
JO - BBA Section Nucleic Acids And Protein Synthesis
JF - BBA Section Nucleic Acids And Protein Synthesis
SN - 0005-2787
IS - 4
ER -