Levan is produced by a species of Bacillus in a tightly controlled fermentation of sugar .Aims of this work to valorize agro waste in producing levan (the fructooligosaccharide) and optimize fermentation conditions for microbial production of levan polymer using respons surface methodology. The variables studied were pH, temperature, inoculum size, incubation period and light using sugar cane molasses as substrate and under the optimal environmental condition determined in this
experiment, we conducted another experiment to study the effect of another two variables tween 80 surfactant concentration and the dry weight of bagasse using sugar cane bagasse as substrate . Also to purify levan from Bacillus sp culture media, characterization the levan polymer by H1 NMR spectroscopy and physicochemical properties and to estimate it as glucose using colorimetric phenol sulphuric acid method.