Alkaline phosphatase (E C 3.1.3.1) belongs to the class of
hydrolases and catalyzes the alkaline
hydrolysis of a number of
phosphoric acid esters,
nucleotides etc.
Alkaline phosphatase was produced from
Bacillus spp, isolated from
soil samples. The
Bacillus spp. was identified by
staining and standard biochemical tests after which
screening was done using modified Pikovoskaya’s
agar method.
Production of
alkaline phosphatase using different
substrates like
calcium phosphate along with
casein,
starch,
glucose and
glutamic acid was carried out. High activity was found in
calcium phosphate along with the
casein. The specific activity of the
crude extract was found to be 0.825U and it was subjected to purification by
DEAE-Cellulose ion exchange chromatography. Finally,36% recovery was obtained. The
molar mass was estimated by using 10%
SDS-PAGE and was found to be approximately 84 KD. The optimum activity was at
pH 8.8 and
temperature of 650C.
Alkaline phosphatase activity was enhanced by Mg2++ upto 66% and 80% activity was inhibited by
EDTA.
Alkaline Phosphatase activity was also confirmed by zymography using malachite green
staining method.