Open AccessA Two‐step Synthesis of New Water‐Soluble Polymers of NAD + and ADP
Author(s) -
GOFFIC François,
SICSIC Sames,
VINCENT Christian
Publication year - 1980
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1980.tb04705.x
Subject(s) - nad+ kinase , alcohol dehydrogenase , cofactor , biochemistry , lactate dehydrogenase , dehydrogenase , chemistry , glycerol 3 phosphate dehydrogenase , enzyme , stereochemistry
Alkylation at the N‐1 position of the adenine moiety of NAD + ADP or ATP with 2,3‐epoxypropylacrylatc, followed by polymerization with or without acrylamide at pH 8, gave water‐soluble polymers of NAD + and ADP where the alkyl chain was located at the exocyclic adenine C‐6 amino group. Cofactor incorporations were good to high: 145–447 μmol NAD + /g polymer and 667 μmol ADP/g polymer. About 30%, of the bound NAD + could be reduced with rabbit muscle lactate dehydrogenase, yeast alcohol dehydrogenase and Bacillus subtilis alanine dehydrogenase; 84%, of the bound ADP was phosphorylated with rabbit muscle creatine kinase. High cofactor activities were obtained with polymerized NAD + with alcohol dehydrogenase as enzyme: the initial rate of NAD + polymer reduction was 35–81% that of free NAD + . These values remained substantially high with agarose‐immobilized alcohol dehydrogenase (15–36%) and should eventually allow their use in continuous enzymatic reactors. Enzymatic phosphorylation of ADP polymer by creatine kinase gave an ATP polymer with high biological activity: 480 μmol ATP/g polymer were transformed with yeast hexokinase.