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To maximize biomass production of an ochratoxigenic mould–controlling strain of Lachancea thermotolerans employing response surface methodology (RSM).Using Plackett–Burman screening designs (PBSD) and central composite designs (CCD), an optimized culture medium containing (g l−1): fermentable sugars (FS), 139·2, provided by sugar cane molasses (CMz), (NH4)2HPO4 (DAP), 9·0, and yeast extract (YE), 2·5, was formulated. Maximal cell concentration obtained after 24 h at 28°C was 24·2 g l−1cell dry weight (CDW). The mathematical model obtained was validated in experiments performed in shaken-flask cultures and also in aerated bioreactors. Maximum yield and productivity values achieved were, respectively, of 0·23 g CDW/g FS in a medium containing (g l−1): FS, 87·0; DAP, 7·0; YE, 1·0; and of 0·96 g CDW l−1 h−1 in a medium containing (g l−1): FS, 150·8 plus DAP, 6·9.Optimized culture conditions for maximizing yeast biomass production determined in flask cultures were applicable at a larger scale. The highest yield values were attained in media containing relatively low-CMz concentrations supplemented with DAP and YE. Yeast extract would not be necessary if higher productivity is the aim.Cells of L. thermotolerans produced aerobically could be sustainably produced in a medium just containing cheap carbon, nitrogen and phosphorus sources. Response surface methodology allowed the fine-tuning of cultural conditions.