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Biosynthesis of extracellular inulinase by bacteria Bacillus polymyxa 29, B. polymyxa 722, and B. subtilis 68 was studied. The optimal parameters for the producer growth were as follows: pH 7.0, 33–35°C, growth duration 72 h. The presence of reduced mineral nitrogen or organic nitrogen was necessary for the enzyme biosynthesis. The inulinase biosynthesis was sharply activated in the presence of carbohydrates. B. polymyxa 722 and B. polymyxa 29 displayed the maximum activities on a starch-containing culture medium; the maximum activity of B. subtilis 68 was in the presence of sucrose. Inulin did not induce inulinase biosynthesis by the strains studied. The time course of bacteria growth and enzyme biosynthesis was studied.