植物種子発芽体の核酸分解酵素に関する研究 : (第2報)緑豆発芽体抽出液の酵素的性質

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By using an enzyme solution extracted from mung bean sprouts, the general properties and their variations of nuclease were studied. Furthermore, a new method for the degradation of RNA was discussed.1) The optimum pH of the nucleases was found in both acidic (pH 4.5-5.0) and alkaline (pH 7.5) regions acting upon RNA, DNA, thymidine-5'-p-nitrophenyl phosphate and bis (p-nitrophenyl) phosphate. From this aspect, these nucleases were assumed to have no substrate specificity.2) The optimum temperature was 60℃ for both acidic and alkaline RNase, They were activated by Mg^>++<, Ca^<++>, Mn^<++> and Zn^<++> and their activities were inhibited by a few chelate compounds and reducing agents. Therefore, it can be concluded that they are metal ion requiring enzymes.3) By the preheating of a solution of the enzyme with 10^<-3>M of Zn^<++> and F^- and with pH at 5.6,at 60℃ for 15 min, the activities of both acidic and alkaline RNase were increased remarkably while the activities of phosphatase were lost by approx. 80%.4) By the preheating described above, the following changes in the enzymatic properties were found : The optimum pH of acidic RNase shifted from 4.5 to 5.0. The optimum temperature of both acidic and alkaline RNase shifted from 60℃ to 80℃. Both acidic and alkaline RNase became more stable against heat and acid.5) The hydrolysate of RNA from these enzymes was identical with 5'-mononucleotides in view of its eluting position in the column chromatogram, maximum ultraviolet absorption spectra, carbazole reaction, periodic Schiff's reaction, recovery of liberated orthophosphorus with 5'-nucleotidase and paperchromatography.6) RNA is hydrolyzed completely to nearly an equal molecule of 5'-mononucleotides by the combined action of acidic and alkaline RNase in a solution of the enzyme. That is, the degradation is carried out by shifting pH of a reaction medium from the acidic to alkaline region or vice versa.
社団法人日本生物工学会の論文
1969-01-25

植物種子発芽体の核酸分解酵素に関する研究 : (第2報)緑豆発芽体抽出液の酵素的性質

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