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

BMC Biotechnology

Marta V Semkiv¹  Kostyantyn V Dmytruk¹  Charles A Abbas²  Andriy A Sibirny³ 

[1] Institute of Cell Biology, NAS of Ukraine, Drahomanov Street, 14/16, Lviv 79005, Ukraine

[2] Archer Daniels Midland Co Research Center, 1001 N Brush College Rd, Decatur, IL 62521, USA

[3] University of Rzeszow, Zelwerowicza 4, Rzeszow 35-601, Poland

关键词: Biomass accumulation;    ATP content;    Alkaline phosphatase;    PHO8;    Ethanol production;    Baker’s yeasts;   

Others  :  834850
DOI  :  10.1186/1472-6750-14-42

 received in 2014-03-06, accepted in 2014-05-07,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 Semkiv et al.; licensee BioMed Central Ltd.

【 图 表 】

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