Pulcherrimin, a red iron chelate, is produced by some yeasts and bacteria. It plays important ecological roles in many ecosystems, including growth control, biofilm inhibition and photoprotection. In this study, fifteen yeast strains of the genus Metschnikowia were characterized based on their production of pulcherrimin. Yeast pulcherrimin was isolated and its purity assessed using 1H nuclear magnetic resonance spectroscopy. Under experimental conditions, pulcherrimin formation varied depending on both the tested strains and culture media. The best producers formed up to 240 mg/L of pulcherrimin in minimal medium with glucose as the carbon source, supplemented with 0.05% FeCl3 and 0.1% Tween 80. This study presents a new approach to producing high yields of pulcherrimin from yeasts.