Purification and characterization of a new ribopolynucleotide synthesizing enzyme from Escherichia coli.

A new type of ribopolynucleotide-synthesizing enzyme was found both on cytoplasmic membranes and in protein-DNA complexes isolated from Escherichia coli. The enzyme was purified by exploiting a specific, reversible enzyme aggregation with ATP and spermidine. The purified enzyme (more than 90% pure) was free from other enzymatic activities such as ATPase and polynucleotide phosphorylase. The enzyme (molecular weight 270,000 ± 15%) contains two kinds of polypeptide chain (molecular weights 91,000 ± 10%, and 45,000 ± 10%) and these polypeptides are not common with those of DNA-dependent RNA polymerase. The enzyme catalyses the synthesis of ribopolynucleotides from nucleoside triphosphates in the presence of 1 mm-MgCl₂. Rifampicin and streptolydigin do not affect the enzyme reaction.