The co-cultivation of GRAS amylolytic bacteria together with ethanol-producing yeast Saccharomyces cerevisiae in starch-containing media might be one of the ways solving the problem of starch-containing waste disposal with simultaneous formation of ethanol as a potential biofuel for increasing octane number of gasoline. The aim of the study was to test the combination of microorganisms (amylolytic lactic acid bacteria and yeast) suitable for co-cultivation on starch and to optimize the conditions for starch cofermentation. Methods. Conventional microbiological, biochemical and statistical methods, including serial dilution technique with counting colony forming units (CFU) for growth assessment of mixed cultures, Gas Chromatograph/Mass Spectrometer (GC/MS) for measuring ethanol concentration and Box-Behnken experimental design (Statistica 10) for bioethanol production optimization, were used in this work. Results. The combination of microorganisms for mixed cultures co-cultivation in single-stage starch fermentation was established: the strain of ethanol-producing yeast S. cerevisiae UCM Y-527 and the amylolytic strain of lactic acid bacteria Streptococcus bovis IMV B-7151. Mathema
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