The study aimed to purification of acid phosphatase (ACP) from sera of obesetype 2 diabetes mellitus patients, this study included from thirty T2DM patients and thirty control, purification process was done with several steps included precipitation with inorganic salt (NH4 ) 2SO4 30%-80%, dialysis, ion exchange chromatography by DEAE sepharose anion column and size exclusion chromatography by Sepharose 6B.ACP, BMI, FBS, HbA1c, Lipid profile, Urea, Creatinie, Insuline, Homa-IR were determined. Results showed the precipitate and concentrated protein appeared four peaks in ion exchange column. ACP located in the first and second peak with purification fold (21.1), (37.2) yield of enzyme and specific activity (173.3) IU/ml, which obtained a single peakby gel filtration chromatography, the degree of purification (34.1) fold, yield of enzyme (20.5) with specific activity (280) IU/ml. Also, the peak that has the highest enzymatic activity showed single peak after eluted in gel filtration chromatography following steps by using SDS-PAGE Electrophoresis. From this study, it is concluded acid phosphatase which purification from sera of obese T2DM patients have two isoenzymes, also, concluded the purified enzyme had an optimum temperature (500C) and optimum pH (5). Purity and molar mass was measured using (SDS.PAGE) electrophoresis showing approximately ~ 56 KD with single band.
