Increased expression of the yeast multidrug resistance ABC transporter Pdr18 leads to increased ethanol tolerance and ethanol production in high gravity alcoholic fermentation

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作者：Miguel C Teixeira (1)
Cláudia P Godinho (1)
Tania R Cabrito (1)
Nuno P Mira (1)
Isabel Sá-Correia (1)
关键词：Saccharomyces cerevisiae ; Ethanol tolerance ; ABC multidrug transporters ; Membrane permeabilization ; Bio ; ethanol production
刊名：Microbial Cell Factories
出版年：2012
出版时间：December 2012
年：2012
卷：11
期：1
全文大小：287KB
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作者单位：Miguel C Teixeira (1)
Cláudia P Godinho (1)
Tania R Cabrito (1)
Nuno P Mira (1)
Isabel Sá-Correia (1)

1. IBB -Institute for Biotechnology and BioEngineering, Centro de Engenharia Biológica e Química, and Department of Bioengineering, Instituto Superior Técnico, Technical University of Lisbon, Av. Rovisco Pais, 1049-001, Lisbon, Portugal
ISSN：1475-2859

文摘

Background The understanding of the molecular basis of yeast tolerance to ethanol may guide the design of rational strategies to increase process performance in industrial alcoholic fermentations. A set of 21 genes encoding multidrug transporters from the ATP-Binding Cassette (ABC) Superfamily and Major Facilitator Superfamily (MFS) in S. cerevisiae were scrutinized for a role in ethanol stress resistance. Results A yeast multidrug resistance ABC transporter encoded by the PDR18 gene, proposed to play a role in the incorporation of ergosterol in the yeast plasma membrane, was found to confer resistance to growth inhibitory concentrations of ethanol. PDR18 expression was seen to contribute to decreased 3?H-ethanol intracellular concentrations and decreased plasma membrane permeabilization of yeast cells challenged with inhibitory ethanol concentrations. Given the increased tolerance to ethanol of cells expressing PDR18, the final concentration of ethanol produced during high gravity alcoholic fermentation by yeast cells devoid of PDR18 was lower than the final ethanol concentration produced by the corresponding parental strain. Moreover, an engineered yeast strain in which the PDR18 promoter was replaced in the genome by the stronger PDR5 promoter, leading to increased PDR18 mRNA levels during alcoholic fermentation, was able to attain a 6- higher ethanol concentration and a 17- higher ethanol production yield than the parental strain. The improved fermentative performance of yeast cells over-expressing PDR18 was found to correlate with their increased ethanol tolerance and ability to restrain plasma membrane permeabilization induced throughout high gravity fermentation. Conclusions PDR18 gene over-expression increases yeast ethanol tolerance and fermentation performance leading to the production of highly inhibitory concentrations of ethanol. PDR18 overexpression in industrial yeast strains appears to be a promising approach to improve alcoholic fermentation performance for sustainable bio-ethanol production.

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