1958 年 78 巻 4 号 p. 362-368
Some improvements were effected in the method of Sinsheimer for separating phosphodiesterase from snake venom by acetone fractionation. Application of this improved procedure to snake venom from domestic asp (Agkistrodon halys blomhoffi BOIE) effected separation of phosphodiesterase by a comparatively easy procedure. However, this method cannot be termed suitable considering the increased specific activity of phosphodiesterase, recovery of the enzyme, and utilization of enzymes other than phosphodiesterase.
A method was devised for obtaining phosphodiesterase of high purity in a good yield from asp venom by the use of column chromatography with cellulose-calcium phosphate gel as the adsorption agent. The sample of phosphodiesterase obtained by this method contains non-specific phosphomonoesterase but not 5′-nucleotidase that it can be used for structural studies of nucleic acid. Use of ammonium sulfate fractionation to the mixture of L-amino acid oxidase and lecithinase-A, separated from phosphodiesterase by this column chromatography, afforded L-amino acid oxidase free from lecithinase-A. The purity of L-amino acid oxidase so obtained was about 11 times that of the crude snake venom. The crude snake venom contains a substance that suppresses the action of L-amino acid oxidase and this substance is adsorbed on calcium phosphate gel.