Purification and Characterization of Acid Phosphatase from the Egg of the Lady Beetle, Harmonia axyridis (Coccinellidae: Coleoptera)

초록

Acid phosphatase (AP) in the egg of the lady beetle, Harmonia axyridis, waspurified and characterized. Ammonium sulfate precipitation, CM column and isoelectrofocusing(IEF) were applied to purify an estimated molecular weight of 66 kDa AP. The purity was checkedby SDS PAGE, native PAGE and Western blot. AP was detected in the hemolymph of the femaleand the egg, but not in the male on the blotting. Km of AP for a substrate, p-nitrophenylphosphate (p-NPP), was 1.64×10-4 M. AP had the optimum enzymatic activity at pH 3.5. Ininhibition tests performed with various chemicals, ammonium molybdate suppressed 99% of theenzyme activity of AP even at the concentration of 5×10-4 mM. AP was stable up to 50.C.

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