The objective of this work was to produce and characterizing glucose syrup from enset starch. The production of glucose syrup (fermentable sugar) from enset starch was carried out via acid hydrolysis and enzymatic hydrolysis. In the present study, enset starch was extracted from bulla and the physicochemical properties were determined. The result showed that physicochemical properties of enset starch were 0.164% ash, 0.602 g/cm3 bulk density, 14.173% moisture, and 28.507% amylose content with a 6.7 PH value. The effect of acid concentration, enzyme concentration, and hydrolysis temperature on the physio-chemical properties of glucose syrup (reducing sugar, dextrose equivalent, total solids, ash, and moisture content) was studied. Experiments were carried out at a temperature range of (100-140) ℃ and (0.2-0.8) M strength of H2SO4 acid during acid hydrolysis while (0.025- 0.075) v/v % enzyme concentration (enzyme-substrate ratio) & (55-65) ℃ range of temperature were considered during enzyme hydrolysis for 2-hour operation. All experiments were done in triplicate and analysis of variance (ANOVA) with Fisher LSD test at p≤ 0.05 was used to measure mean difference among treatments. The result revealed that the maximum glucose yield (46.067%), reducing sugar (55.991%), dextrose equivalent (76.898%), and degree Brix (49) were obtained at 120℃ using 0.8M H2SO4 during acid hydrolysis. While the highest glucose yield (56.643%), reducing sugar (64.721%), dextrose equivalent (94.718%), and degree Brix (68.947) were obtained at 60℃ and 0.075% enzyme concentration during enzymatic hydrolysis. The analysis of variance (ANOVA) result showed that the variation of temperature, acid concentration, and enzyme concentration has a significate (p≤0.05) effect on the physio-chemical properties of the produced glucose syrup.