Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast

Anna Judith Schifferdecker, Juozas Siurkus, Mikael Rørdam Andersen, Dorte Jørck-Ramberg, Zhihao Ling, Nerve Zhou, James E. Blevins, Andriy A. Sibirny, Jure Piškur, Olena P Ishchuk

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    Abstract

    Dekkera bruxellensis is a non-conventional Crabtree-positive yeast with a good ethanol production capability. Compared to Saccharomyces cerevisiae, its tolerance to acidic pH and its utilization of alternative carbon sources make it a promising organism for producing biofuel. In this study, we developed an auxotrophic transformation system and an expression vector, which enabled the manipulation of D. bruxellensis, thereby improving its fermentative performance. Its gene ADH3, coding for alcohol dehydrogenase, was cloned and overexpressed under the control of the strong and constitutive promoter TEF1. Our recombinant D. bruxellensis strain displayed 1.4 and 1.7 times faster specific glucose consumption rate during aerobic and anaerobic glucose fermentations, respectively; it yielded 1.2 times and 1.5 times more ethanol than did the parental strain under aerobic and anaerobic conditions, respectively. The overexpression of ADH3 in D. bruxellensis also reduced the inhibition of fermentation by anaerobiosis, the "Custer effect". Thus, the fermentative capacity of D. bruxellensis could be further improved by metabolic engineering.
    Original languageEnglish
    JournalApplied Microbiology and Biotechnology
    Volume100
    Issue number7
    Pages (from-to)3219-3231
    Number of pages13
    ISSN0175-7598
    DOIs
    Publication statusPublished - 2016

    Bibliographical note

    This article is published with open access at Springerlink.com

    An erratum to this article can be found at http://​dx.​doi.​org/​10.​1007/​s00253-016-7349-3.

    Keywords

    • ADH3
    • Auxotrophic mutants
    • Dekkera bruxellensis
    • Gene expression
    • Glucose fermentation
    • Promoters
    • TEF1

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