A technique for the identification and separation of enzymes by paper chromatography

Abstract

Enzyme solns. were chromatogramed on filter paper at 0-5[degree]C using aqueous acetone or 033 [image]M NaCl as solvent. The dried paper was then placed on an agar plate containing substrate. After 4-12 hrs. at room temp. the paper was removed and the agar surface tested for spots (e. g. with I2 for amylases and phosphorylases, 0.1 [image] NaOH for phosphatases). Rp values are given for a number of plant amylases and phosphorylases. Separation of mixed enzyme was satisfactorily achieved. Separations were achieved of the amylases of germinated rice, of kidney phosphatases, of liver acid and alkaline phosphatase, and of serum alkaline and acid phosphatasp. The amylase and phosphorylase of green gram could not be separated.