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Víctor Guadalupe-Medina, Benjamin Metz, Bart Oud, Charlotte M. van Der Graaf, Robert Mans, Jack T. Pronk and Antonius J. A. van Maris Evolutionary engineering of a glycerol-3-phosphate dehydrogenase-negative, acetate-reducing Saccharomyces cerevisiae strain enables anaerobic growth at high glucose concentrations Microbial Biotechnology 7

Version of Record online: 4 SEP 2013 | DOI: 10.1111/1751-7915.12080

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Glycerol formation in anaerobic S. cerevisiae cultures was recently eliminated by expressing Escherichia coli (acetylating) acetaldehyde dehydrogenase (encoded by mhpF) and simultaneously deleting the GPD1 and GPD2 genes encoding glycerol-3-phosphate dehydrogenase, thus coupling NADH reoxidation to reduction of acetate to ethanol. Gpd- strains are, however, sensitive to high sugar concentrations, which complicates industrial implementation of this metabolic engineering concept. In this study, laboratory evolution was used to improve osmotolerance of a Gpd- mhpF-expressing S. cerevisiae strain, resulting in an increase of the ethanol yield on 1M sugar from 79 % of the theoretical maximum in the Gpd+ reference strain to 92 % for the Gpd- evolved strains.

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