Sixty one isolates of Penicillium crustosum originating from various foodstuffs were screened for penitrem A production by thin-layer chromatography. The highest producers of penitrem A (4 isolates) were grown in various liquid media. Skimmed milk (2%)/potato extract (2%)/sucrose (2%) (SPS) medium supported the highest toxin production and P. crustosum Sp 1552 was selected as the best producer of penitrem A. Optimal conditions for the production of penitrem A were to grow the isolate Sp 1552 (stationary) in 100 ml of SPS medium (with 4% sucrose) in 500-ml flasks for 3 weeks at 25 degrees C with initial pH 5.7. The isolation and purification of penitrem A is described. Approximately 246 mg of pure penitrem A was obtained from 200 flasks each containing 100 ml of SPS medium.