The bioconversion of octanoic acid into 2-heptanone by spores of Penicillium roquefortii is performed using a fed-batch technique with pH control by addition of the liquid substrate itself. The early stage of this process takes place with a high bioconversion rate and high yield. These values then decrease as a result of germination and growth of the biocatalyst. An optimization strategy for the process would thus be to improve the characteristics of this first period, i.e., increase its duration and the reaction rate. An increase in duration is evidenced in two cases : (i) under oxygen limitation; and (ii) when the spore content in the medium is less than 10(7) spores/mL. These conditions give insufficient overall bioconversion rates; better optimization should be achieved without oxygen limitation and with high spore content. Characterization of the first period by material and bioenergetic balances suggests that an increase in the ethanol content of the medium, which acts as an energy source and a permeabilizer, and the use of a specific inhibitor of the Krebs cycle, may be a way to further improve the biocatalyst performance and stability.