Shrimp abdomenal muscle NADP-dependent malic enzyme (E.C.1.1.1.40) was purified about 1500-fold to a specific activity of 48 units (μmol/min)/mg at 30°C with good quantitative recovery in three chromatographic steps, including affinity chromatography on 2′,5′-ADP-Sepharose 4B, a "substrate activation" method using malate substrate plus manganese chloride. In addition to the malate-manganese chloride substrate pair, succinate or glutamate plus manganese chloride or magnesium chloride could be used in this "substrate activation" method for crustacean NADP-malic enzyme purification on 2′,5′-ADP-Sepharose 4B. Affinity chromatography alone purified malic enzyme almost 43 fold, and the overall method resulted in homogeneous enzyme since polyacrylamide gel electrophoresis of the native purified enzyme revealed only a single band staining for protein and enzyme activity.

Additional Metadata
Persistent URL dx.doi.org/10.1016/S0021-9673(01)94454-1
Journal Journal of Chromatography A
Citation
Skorkowski, E.F. (Edward F.), & Storey, K. (1987). Affinity chromatography on 2′,5′-ADP-sepharose 4B for purification of malic enzyme from crustacean muscle. Journal of Chromatography A, 389(C), 427–432. doi:10.1016/S0021-9673(01)94454-1