Purification and isolation of 2 phosphatases from sheep liver

Abstract

The levels of purífication of phosphatases (ortophosphoric monoester phosphydrolases, 3.1.3.1, 3.1.3.2) from the liver were much below those obtained from other sources. A method is described which ineludes as a fundamental step the treat- ment with ion-exchange adsorbents (like to Dowex 50 and Dowex 3), a great quantity of inert protein being retained in the anionic resin. The resulting solution is precipitated by (NJD2SO4 between 45-85 % saturation, redisolved and then precipitated with acetone (60 %). Dissolved in water it is cro- matografied on paper (ethanol-water, 1 : 1), where two frac- tions are distinguished (Rr 0.48 and 0.37) with optimal pH 9.8 and 6 respectively, which can be recovered by elution

Preparations of alkaline phosphatase are by this way obtained with optimal 9.8, specific activity 28.000 units (/xg P/mg N/min) and of acid phosphatase with optimal 6, specific activity 33.000 units. The purification is with respect to theinitial homogenate of some 600 and 1.000 times respectively.