This work reports the use of response surface methodology to optimize the lipase production by submerged fermentation using immobilized cells of a newly isolated Penicillium sp. For this, experimental designs were employed to evaluate the best condition for microorganism immobilization. The optimum experimental conditions for immobilized microorganisms were determined as 2.5% (w/v) of calcium alginate, 0.2% (w/v) of CaCl2, and 1.5% (v/v) of glutaraldehyde and 6 h of curing time, yielding a lipase activity as high as 20.96 U/mL in 120 h of fermentation. The activity obtained is higher than those observed with free cells of this microorganism and comparable to results presented in the literature for lipase production with immobilized cells from several microorganisms. The crude enzymatic extract presented Optimum pH and temperature of 7.0 and 37 degrees C, respectively, and maintained its activity for 60 days storage at -10 degrees C.