Abstract: Amylase was purified from the juice of healthy and diseased sugarcane, Ishurdi (Isd)-20 by successive chromatographies on DEAE-Sephadex A-50 and Sephadex G-150 to a homogeneous state as confirmed by polyacrylamide disc gel electrophoresis (PAGE). The molecular weights of the enzyme from healthy and diseased sugarcane juices were estimated as 55.8 and 56.3 kDa, respectively by gel filtration. The purified amylases were of -type and not a glycoprotein in nature. The purified amylases showed the following characteristics, respectively; Km values- 0.27% and 0.23%; optimum pH 6.4 and 6.3; and optimum temperature 39 and 40°C for starch as substrate. Ca2+, Mg2+, and Mn2+ increased amylase activities remarkably. The activities of amylases were increased slightly by Zn2+. On the other hand Cd2+ and Fe3+ increased moderately. Na+ had almost no effect on the amylase activity. Ag+ and K+ produced a little inhibitory effect but Cu2+ inhibited amylase activities moderately. EDTA and acetic acid almost completely ceased amylase activities. Both amylases were found to contain 70% activity in presence of 8M urea.