LAUSR

β-Glucosidase was selected to be a reporter to study metabolic burden imposed by its expression in yeast. Cell growth, fermentation yield and enzymatic activity were used as indicators of the metabolic burden borne by 14 recombinant yeast strains. Various factors were found to affect metabolic burden, including BGLI gene source, gene dose, trafficking of the enzyme (either cell-surface display or secretion), and oxygen supply. While BGLI gene from Aspergillus aculeatus provided better performance for the host cells than that from Saccharomycopsis fibuligera, displaying β-glucosidase on the cell surface generally led to lower μm, total activity and ethanol titer, and longer lag period, lower (aerobic condition) or higher (anaerobic condition) biomass yield than that of secreting β-glucosidase. The negative effect on growth increased with gene dose level until a final failure to grow. This growth difference implies that displaying β-glucosidase on the cell surface imposes an extra metabolic burden. The molecular basis and mechanisms for this phenomenon need to further be investigated in order to develop better strategies for utilizing displayed and secreted enzymes in biotechnology and yeast breeding.