燐酸ジエステラーゼに就いて : 第I報 ジフェニール燐酸を特殊的に水解するジエステラーゼの分離
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概要
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Phosphodiesterase specific for the hydrolysis of diphenylphosphate, one of aromatic phosphodiesters, was obtained from hog kidney by following procedure. To hog kidney homogenized in a Waring blendor with 3 volumes of distilled water was added solid sodium chloride to a final concentration of 1% and after adjusting of pH to 5 with ca. 5 N hydrochloric acid, it was shaken for 10 minutes with an equal volume of n-butanol, allowed to stand for one hour, and centrifuged. The aqueous extract present beneath the floating gel layer of protein and butanol mixture, was siphoned out and fractionated with ammonium sulfate. The precipitate obtained between 45 and 70 % saturation was dissolved in a small volume of distilled water, dialyzed for 48 hours against running tap water, and then, to inactivate the monoesterase, heated in water bath of 100° for 5 minutes, whereby the solution became faintly turbid. The enzyme solution, thus prepared, hydrolyzed diphenylphosphate, liberating phenol but no inorganic phosphate, while it was inactive to monophenylphosphate. Diphenylphosphate of a final concentration of 0.0005M could be hydrolyzed in one hour at the optimum pH 7 to the extent of 40%. At this pH, the enzyme activity was not influenced by addition of Mg^<++>, Ca^<++>, or CN^<->. Bis-dichloroisopropylphosphate, dibenzylphosphate, diethylphosphate, lysolecithin, RNA, and DNA were resistant to this enzyme. It is conceivable that these diesters are hydrolyzed by other enzymes different from the aromatic phosphodiesterase, presented in this paper.
- 千葉大学の論文
- 1958-05-28