Abstract: Acid phosphatase (ACPase, EC3.1.3.2) was isolated and purified from the Italian honeybee, Apis mellifera L., and properties of the enzyme had been studied. Acid phosphatase was partially obtained from A. mellifera by homogenate, a mmunium sulfate fractionation, chromatography with DEAE-sepharose FF and gel filtration with Sephadex G-200 The purified enzyme moved as a single electrophoretic band in PAGE. The purification multiple was 77.24, and the specific activity 16.22 U/mg with pNPP as its substrate. The molecular weight of ACPase was 135 kD determined with gel filtration and its subunit weight was 63.1 kD determined with SDS-PAGE. Isoelectric focusing study showed that pI values of the enzyme were 4.46 and 4.79. NR/R single and two dimensions SDS-PAGE indicated that the enzyme contained intrachain disulfide bond. Circular dichroism spectrum was investigated, and it was found that the proportion of α-helix, β-sheet and random coil in the enzyme was 13.84%, 2.68% and 6.34% respectively. Amino acid composition analysis showed that there were about 507 amino acids in the ACPase, with plenty of Asp.

Key words: Apis mellifera, acid phosphatase, isolation and purification, properties