Die Darstellung und Reinigung der alkalischen Phosphatase aus Pferdenieren. 1. Mitteilung

Abstract

Fresh horse kidneys were freed from fat and skin and ground in a meat grinder. To 2 kg. of kidney were added 2 1. of 50% alcohol, 100 cc. toluol and 100 cc. of ethyl acetate. The mixture was autolyzed for 4 days at 35[degree]-38[degree], and stirred once each day. The mixture was filtered and 2 l. of the ice cold filtrate adjusted to pH 4.4 by the addition of 4 cc. of acetic acid. After removal of the slight ppt. formed, 2 1. of absolute alc. denatured with petroleum ether were added, and the soln. placed in the ice box for 1 hr. The ppt. formed was filtered off, washed with 100 cc. absolute alc. and 100 cc. ether. The residue was dried in a vacuum desiccator over CaCl2, and formed a light gray powder. The phosphatase activity of the substance was detd. as mgms. of P2O5 liberated per hr. at 35[degree], using Na-[beta]-glycero-phosphate as substrate. The enzyme prepared from horse kidney showed greater activity than that prepared from human or swine kidney.