Continuous Production of Homopolynucleotides by Immobilized Polynucleotide Phosophorylase

概要

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A polynucleotide phosphorylase was immobilized with glutaraldehyde, via an aminopropyl spacer, on porous glass. The specific activity of the immobilized enzyme was effectively increased by the addition of an appropriate ribonucleoside diphosphate on immobilization.A homopolynucleotide could be synthesized continuously by passing a nucleoside diphosphate solution through the immobilized enzyme column. The chain length of the product depended upon the temperature and the flow rate. Polyinosinic acid, poly(I), was continuously synthesized with the immobilized enzyme for about one month without appreciable loss of activity. Polyinosinic acid-polycytidylic acid, poly(I)・poly(C), prepared from poly(I)and poly(C)synthesized with the immobilized polynucleotide phosphorylase, induced interferon-β(IFN-β)in human cultured cells as effectively as that prepared from homopolynucleotides synthesized with the free enzyme.
社団法人日本生物工学会の論文
1986-12-25