Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes

Figure 1

Size-exclusion chromatography of intracellular sucrase. The ammonium sulfate saturated sample protein was run on a Sephacryl (S-200) column, preequilibrated with Buffer A. The protein absorbance measured at 280 nm (--) was plotted on primary axis. The secondary -axis shows the intracellular sucrase activity (--) of the corresponding fractions. The molecular weight of the native enzyme was determined to be ~112 kDa from the S-200 calibration graph (Figure 1 inset). The arrow indicates the peak of intracellular sucrase elution volume and the molecular weight (MW) of the protein. The molecular weight markers used were as follows: amylase MW = 200,000; alcohol dehydrogenase MW = 150,000; hemoglobin MW = 64,500; carbonic anhydrase MW = 29,000; and cytochrome C MW = 12,400.