Our research group developed an efficient protocol to pro-duce nisin from L. lactis, with whey milk as culture medium. This by-product from dairy industry isa low-cost alternative for industrial production, which canprovide high yields of this antimicrobial agent. To our knowl-edge, there are no reports on nisin purification from thismedium. In fact, all methods described for nisin purifica-tion used complex formulated media, mostly Man, Rogosa,and Sharpe (MRS) medium. The aim of the present workwas to purify produced nisin through hydrophobic interac-tion chromatography (HIC), based on purification profiles ofcommercial nisin.