Biofilm-based fermentation: a novel immobilisation strategy for Saccharomyces cerevisiae cell cycle progression during ethanol production.

Biofilm-based fermentation, as a new immobilisation strategy, is beneficial for industrial fermentation due to its excellent environmental resistance, high productivity and continuous fermentation relative to calcium alginate-immobilised fermentation. These two techniques differ mainly regarding cell stages. Here, we describe the cell phenotype of Saccharomyces cerevisiae biofilm-based fermentation and compare cell cycle stages with those during immobilisation in calcium alginate. Most cells in the biofilm-based fermentation adhered to the cotton-fibre carrier of the biofilm and were in the G2/M phase whereas alginate-embedded cells were in the G1/G0 phase. Deletion of the RIM15 gene, which regulates cell cycle progression according to nutritional status, hampered the cell cycle arrest observed in alginate-embedded cells, enhanced biofilm formation and improved fermentation ability. The improved biofilm formation shown by the rim15△ strain could be attributed to an increase in the expression level of the adhesion protein FLO11 and synthesis of trehalose. These findings suggest that the extracellular environment is mainly responsible for the difference between biofilm-based fermentation and alginate-embedded fermentation, and that RIM15 plays an essential role in cell cycle progression. KEY POINTS: • In the biofilm, S. cerevisiae cell populations were mostly in the G2/M phase while alginate-embedded cells were arrested in the G1/G0 phase. • The RIM15 gene partially influenced the cell cycle progression observed during ethanol fermentation. • Biofilm-based cells were actively adsorbed on the physical carrier. • Biofilm immobilisation could maintain cell division activity explaining its fermentation efficiency.