The strain of Bacillus licheniformis LBA 46 (from the culture collection of the Laboratory of Food Biochemistry, School of Food Engineering, UNICAMP) was cultivated in a 6 L bench reactor using 32 g/L of sugar cane molasses, Fio de Ouro®, BR; 6 g/L of corn steep liquor, Corn Products Brasil; 2 g/L of yeast extract, Prodex–Lac SD® Produtos Especiais para Alimentos S/A; and 20 g/L dried whey, Alibra, pH 7. The protease was produced under the following fermentation conditions: 48h, 30 °C and 300 rpm. The supernatant was separated by centrifugation and fractionated with 80% ammonium sulfate. The precipitate was dissolved in 0.05 M phosphate buffer pH 7.0 and dialyzed against distilled water at 5 °C. The dialyzed protease was applied to a 20 mL DEAE Sepharose column (HiPrep DEAE FF 16/10, GE) equilibrated with the same buffer. The proteins were eluted (5 mL/min) using 0.05 M phosphate buffer pH 7.0 containing a linear (0 to 1 M) sodium chloride gradient (Äkta Purifier, GE), and active fractions which showed protease activity were pooled. The molecular weight of the protease was estimated by SDS–PAGE. The effect of pH (4 – 11) and temperature (30 – 80 ºC) on the protease activity and stability was determined using a univariate assay. The protease was purified 3.30 fold after the ammonium sulfate precipitation and DEAE Sepharose column chromatography. The purified protease showed a specific activity of 628.96 U/mL and the molecular weight was estimated as 40 kDa by SDS–PAGE. The purified protease presented stability between pH 5 – 10 and between 30 – 50 ºC (activity higher than 80%) with the optimum at pH 8.5 and 60 ºC. The purified protease was shown to be stable over a wide range of pH and temperature values, which is an interesting feature for its biotechnological use.