Purificacion de fosfatasa acida de higado de bovino

Several mammalian tissues have been shown to contain
as many as three acid phosphatases which may be separated
on the basis of differences in size by gel filtration on Sephadex
G-75. The smallest of these enzymes from bovine liver,
termed acid phosphatase III, has been purified to apparent
homogeneity. The purified enzyme migrates as a single
band during electrophoresis on polyacrylamide gel and
yields arginine as the only detectable amino terminal residue.
Acid phosphatase III has been characterized with respect to
its substrate specificity, molecular weight, and amino acid
composition. The K, value determined with p-nitrophenyl
phosphate, the most readily hydrolyzed substrate tested, is
7.5 X 10u4 M. Optimal rates of hydrolysis were observed
at pH 5.5.
The pure enzyme is stabilized by ethylenediaminetetraacetate
and phosphate ion, but is rapidly inactivated
in the presence of Mg++ or mercaptoethanol. The
molecular weight of acid phosphatase 111 determined by
sedimentation equilibrium analysis is 16,590. This figure
is in close agreement with the value of 16,296 obtained from
the amino acid composition.

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