Recently, inulinases one of the most important enzymes which have gained much attention in industrial purposes. They can be applied for production of high fructose syrup from inulin and could reduce the process steps, resulting in cost reduction and increasing the amount of products. Acetobacter sp. produced extracellular inulinase  in vitro. Maximum inulinase production was obtained at pH 7.0, 30⁰C temperature, ammonium sulphate as nitrogen source, inoculum volume 5% (v/v), 1% sucrose as additional carbon source and 48 hr after inoculation.The enzyme showed maximum activity at pH 7 and 50 ⁰C. The enzyme was stable more than 80% of its maximal activity at 30-50 ⁰C after 30 min heat treatment.