Few strains of Metschnikowia pulcherrima (Pitt) M.W. Miller are under development for control of postharvest pathogens on fruit. A substrate was developed to optimize the biomass production of M. pulcherrima strain BIO126. Different complex nutrient sources, with or without pH control, were tested. Growth in yeast extract provided at concentrations > or =30 g*L-1 yielded the highest biomass. The addition of two carbon sources, D-mannitol and L-sorbose, at 5 g*L-1 each, significantly improved yeast growth. The greatest amount of yeast growth occurred when pH values of the medium ranged from 5.0 to 7.5. A combination of yeast extract, D-mannitol, and L-sorbose (YEMS), probably with diauxic utilization, showed a synergistic effect, widening the exponential phase (maximum specific growth rate of 0.45 h-1) and increasing the final cell number (1.5 x 109 cells*mL-1) and dry biomass (6.0 g*L-1) in well-controlled batch fermentation. In efficacy trials on 'Golden Delicious' apples, M. pulcherrima grown in YEMS effectively reduced incidence and severity of Botrytis cinerea (51.1% and 70.8%, respectively) and Penicillium expansum (41.7% and 14.0%, respectively). Also on 'Gala' apples, the best reduction of grey and blue mould incidence was obtained with cells grown in YEMS (58.1% and 50.5%, respectively).