The bacteriocin pediocin PA-1 produced by Pediococcus acidilactici PAC 1.0 offers significant potential as a food preservative and as an antimicrobial agent in the medical area. However, low production yields and difficulties in obtaining significant amounts of pure pediocin PA-1 have limited, in part, its biochemical and physical characterization. In the present study, we describe a simple and more efficient purification strategy for pediocin PA-1. A hydrophobic interaction chromatography step using an octyl-Sepharose column was introduced for final purification and polishing. The new method is a scalable one, uses only two steps and yields highly purified pediocin PA-1 with a recovery as high as 73%, which is at least two to three times more than that of the methods reported so far. Highly purified, biologically active pediocin PA-1 of the correct molecular mass (4624 Da, with two disulphide bridges) was obtained. Fourier-transform infrared analysis runs at p2H 6 indicated that pediocin PA-1 was more structured than similar pediocin PA-1 samples purified using the earlier purification scheme.