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Background  The production of ethyl alcohol by fermentation represents the largest scale application of  Saccharomyces cerevisiae  in industrial biotechnology. Increased worldwide demand for fuel bioethanol is anticipated over the next decade and will exceed 200 billion liters from further expansions. Our working hypothesis was that the drop in ATP level in  S. cerevisiae  cells during alcoholic fermentation should lead to an increase in ethanol production (yield and productivity) with a greater amount of the utilized glucose converted to ethanol. Our approach to achieve this goal is to decrease the intracellular ATP level via increasing the unspecific alkaline phosphatase activity.    Results  Intact and truncated versions of the  S. cerevisiae PHO8  gene coding for vacuolar or cytosolic forms of alkaline phosphatase were fused with the alcohol dehydrogenase gene ( ADH1 ) promoter. The constructed expression cassettes used for transformation vectors also contained the dominant selective marker  kanMX4  and  S. cerevisiae  δ-sequence to facilitate multicopy integration to the genome. Laboratory and industrial ethanol producing strains BY4742 and AS400 overexpressing vacuolar form of alkaline phosphatase were characterized by a slightly lowered intracellular ATP level and biomass accumulation and by an increase in ethanol productivity (13% and 7%) when compared to the parental strains. The strains expressing truncated cytosolic form of alkaline phosphatase showed a prolonged lag-phase, reduced biomass accumulation and a strong defect in ethanol production.    Conclusion  Overexpression of vacuolar alkaline phosphatase leads to an increased ethanol yield in  S. cerevisiae .

bmc biotechnology

Increased ethanol accumulation from glucose via reduction of ATP level in a recombinant strain of Saccharomyces cerevisiaeoverexpressing alkaline phosphatase