Purification and Properties of 5'-Nucleotidase from Cod Muscle(Biological Chemistry)

概要

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An enzyme which catalyzes the hydrolysis of 5'-IMP to inosine and inorganic phosphate was purified from cod muscle by extraction of the acetone powder with 1% Triton X-100, and followed by two steps of affinity chromatography on Concanavalin A-Sepharose and 5'-AMP Sepharose 4B. The purified enzyme was homogeneous on disc polyacrylamide gel electrophoresis. The apparent molecular weight of the purified enzyme was estimated to be 2.7 × 10^5 by gel filtration, and about 6.7 × 10^4 by SDS-polyacrylamide gel electrophoresis. The enzyme was identified as 5'-nucleotidase (EC 3.1.3.5), since it hydrolyzed various 5'-nucleotides but not a number of phosphorylated compounds including 2'(3')-nucleotides. The optimum pH was 7.6, and there appeared a second peak at pH 9.1 in the presence of Mg^<2+>. The enzyme was inhibited by Zn^<2+> and Cu^<2+>, but was not inhibited by Ca^<2+>, Ba^<2+>, 2-mercaptoethanol, pCMB and inosine. The enzyme was competitively inhibited by ADP and ATP, and was also inhibited by EDTA, depending upon the concentration of EDTA and the incubation time. The EDTA-inactivated enzyme was partially reactivated by the addition of some divalent cations, Zn^<2+> being the most effective.
社団法人日本農芸化学会の論文
1986-05-23