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Purification and Characterization of Acid Phosphatase from the Egg of the Lady Beetle, Harmonia axyridis (Coccinellidae: Coleoptera)
Author(s) -
LEE Jun Hyuk,
PARK Yong Chul
Publication year - 2004
Publication title -
entomological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.421
H-Index - 20
eISSN - 1748-5967
pISSN - 1738-2297
DOI - 10.1111/j.1748-5967.2004.tb00085.x
Subject(s) - biology , coccinellidae , ammonium sulfate precipitation , acid phosphatase , hemolymph , western blot , microbiology and biotechnology , harmonia axyridis , botany , enzyme , chromatography , biochemistry , ecology , chemistry , size exclusion chromatography , gene , predation , predator
Acid phosphatase (AP) in the egg of the lady beetle, Harmonia axyridis , was purified 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 checked by SDS PAGE, native PAGE and Western blot. AP was detected in the hemolymph of the female and the egg, but not in the male on the blotting. Km of AP for a substrate, p ‐nitrophenyl phosphate ( p ‐NPP), was 1.64 x 10 ‐4 M. AP had the optimum enzymatic activity at pH 3.5. In inhibition tests performed with various chemicals, ammonium molybdate suppressed 99% of the enzyme activity of AP even at the concentration of 5 x 10 ‐4 mM. AP was stable up to 50°C.