Batch and Fed-Batch Ethanol Fermentation of Cheese-Whey Powder with Mixed Cultures of Different Yeasts

Eight yeast strains of Lachancea thermotolerans, Kluyveromyces marxianus, and Kluyveromyces waltii have been tested for their ability to ferment lactose into ethanol in mashes containing 10% (w/v) cheese whey powder (CWP). The K. marxianus NCAIM Y00963 achieved 3.5% (v/v) ethanol concentration at 96–120 h of fermentation. The ethanol production by the selected lactose-positive strains and the well-known ethanologenic Saccharomyces cerevisiae (Levuline Fb) in mixed culture was also investigated at different CWP concentrations and inoculation techniques in batch mode. The mixed culture in an equal ratio (1:1) of cell counts of K. marxianus and S. serevisiae showed an increase in lactose conversion rate. The two yeast strains in a ratio of 3:1 (three-quarters of K. marxianus and a quarter of S. cerevisiae in a total of 4.5 × 1010 cells) resulted in 72.33% efficiency of lactose bioconversion and 7.6% (v/v) ethanol production at 17.5% (w/v) of CWP concentration. In the repeated inoculation process, with the addition of three-quarter part of 3:1 ratio of mixed culture (3.3 × 1010 cells of K. marxianus) into 150 mL CWP mash at initiation and the rest quarter part (1.2 × 1010 cells of S. cerevisiae) at 24 h, 8.86% (v/v) ethanol content with 87.5% efficiency of lactose conversion was reached. Both the ethanol concentration and efficiency of bioconversion were increased to 10.34% (v/v) and 92%, respectively, by combination with fed-batch fermentation technology. Our results can serve a very good basis for the development of industrial technology for the utilization of cheese whey.