BACKGROUND: The present work aims to gain further insight on the use of Fusarium solani pisi cutinase from a Saccharomyces cerevisiae strain for the synthesis of short chain ethyl esters, a group of important fruit flavor compounds, in a non-conventional environment. Synthesis is promoted by cutinase in organic media, in particular, in iso-octane, an organic solvent recognized as a safe ingredient in food and beverage industrial processes. RESULTS: The effect of solvent and substrate components of the reaction mixture on the enzyme stability was measured separately. Focus was given to the effect of reaction medium on the operational stability of cutinase and on the esterification yield after 24h. The feasibility of the operation in fed batch mode was successfully evaluated. The effect of the addition of substrate in consecutive pulses in the activity and stability of the biocatalyst was again assessed. CONCLUSIONS: The bioconversion system used in this work allowed for the sustained production of short chain alkyl esters for more than 45 days, which is suggestive of the stability of cutinase in the organic environment evaluated. (C) 2010 Society of Chemical Industry