A strain of Lactobacillus plantarum was examined for production of an extracellular antagonistic compound. Cellfree preparations, dialyzed to remove organic acids, were used in inhibition studies which revealed that Gram-positive bacteria were sensitive. Among these, Staphylococcus aureus was chosen for further characterization of the agent. The antagonistic compound was susceptible to breakdown by proteolytic enzymes and its effect was completely lost after heat treatment at 121°C for 15 min. Ultrafiltration studies indicated that the agent had a molecular weight of over 100, 000, suggesting a complex protein-containing aggregate. The antagonistic effect was found to be highest at low pH values and S. aureus was shown to be able to adapt to the agent. Most Gram-negative bacteria were resistant to the compound. However, after their transformation to spheroplasts, which removed most of the cell envelopes, these bacteria were sensitized. The conclusion is that the antagonistic mechanism probably includes agent influence on the cell surface. © 1986.