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.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-015-7266-x) contains supplementary material, which is available to authorized users.
Highlights
The wine and beer yeast Dekkera bruxellensis is the main cause of wine spoilage worldwide, thereby causing immense economic losses (Boulton et al 1996; Fugelsang 1996; Delfini and Formica 2001; Loureiro and Malfeito-Ferreira 2003; Schifferdecker et al 2014)
Several D. bruxellensis strains were mutagenized by UV or ethane methyl sulfonate (EMS), and thousand colonies of each strain screened for auxotrophic mutants
We developed new molecular biology tools that we used for the metabolic engineering of D. bruxellensis
Summary
The wine and beer yeast Dekkera bruxellensis (anamorph Brettanomyces bruxellensis) is the main cause of wine spoilage worldwide, thereby causing immense economic losses (Boulton et al 1996; Fugelsang 1996; Delfini and Formica 2001; Loureiro and Malfeito-Ferreira 2003; Schifferdecker et al 2014). Infected wines develop distinctive and unpleasant aromas due to volatile phenols produced by this species, called BBrett’ taints^ (Chatonnet et al 1995) and normally associated with the smell of barnyard, burnt plastic, wet animal and horse-sweat (Licker et al 1998). This species is known for its contribution to the flavour composition of Belgium’s Lambic and Gueuze beers (Dequin et al 2003; Dufour et al 2003), specialized red wines (Château Musar and Château de Beaucastel), feta cheese and Kombucha tea (Mayser et al 1995; Teoh et al 2004). Whole genome sequences of 10 Dekkera/Brettanomyces bruxellensis isolates originating from wine, beer or soft drinks were recently reported
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