title: Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast.
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.
authors.affiliation: Department of Biology, Lund University, Sölvegatan 35, Lund, SE-223 62, Sweden. Anna.Schifferdecker@biol.lu.se.
authors.forename: Anna Judith
authors.initials: AJ
authors.lastname: Schifferdecker
authors.affiliation: Department of Systems Biology, Technical University of Denmark, DK-2800, Lyngby, Denmark.
authors.forename: Juozas
authors.initials: J
authors.lastname: Siurkus
authors.affiliation: Department of Systems Biology, Technical University of Denmark, DK-2800, Lyngby, Denmark.
authors.forename: Mikael Rørdam
authors.initials: MR
authors.lastname: Andersen
authors.affiliation: Department of Systems Biology, Technical University of Denmark, DK-2800, Lyngby, Denmark.
authors.forename: Dorte
authors.initials: D
authors.lastname: Joerck-Ramberg
authors.affiliation: Department of Biology, Lund University, Sölvegatan 35, Lund, SE-223 62, Sweden.
authors.forename: Zhihao
authors.initials: Z
authors.lastname: Ling
authors.affiliation: Department of Biology, Lund University, Sölvegatan 35, Lund, SE-223 62, Sweden.
authors.forename: Nerve
authors.initials: N
authors.lastname: Zhou
authors.affiliation: Consulting statistician, Pinnmöllevägen 48, SE-24755, Dalby, Sweden.
authors.forename: James E
authors.initials: JE
authors.lastname: Blevins
authors.affiliation: Institute of Cell Biology, NAS of Ukraine, Drahomanov Street 14/16, Lviv, 79005, Ukraine.|Department of Biotechnology and Microbiology, University of Rzeszow, Zelwerowizca 4, Rzeszow, 35-601, Poland.
authors.forename: Andriy A
authors.initials: AA
authors.lastname: Sibirny
authors.affiliation: Department of Biology, Lund University, Sölvegatan 35, Lund, SE-223 62, Sweden.
authors.forename: Jure
authors.initials: J
authors.lastname: Piškur
authors.affiliation: Department of Biology, Lund University, Sölvegatan 35, Lund, SE-223 62, Sweden. Olena.Ishchuk@biol.lu.se.
authors.forename: Olena P
authors.initials: OP
authors.lastname: Ishchuk
mesh_terms: D000332:Aerobiosis; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000693:Anaerobiosis; D056804:Biofuels; D003001:Cloning, Molecular; D055167:Dekkera / Q000201:enzymology / Q000235:genetics*; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005818:Genetic Engineering; D005947:Glucose / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D056804:Biofuels; D005656:Fungal Proteins; D011994:Recombinant Proteins; D000431:Ethanol; D000426:Alcohol Dehydrogenase; D005947:Glucose
keywords: ADH3; Auxotrophic mutants; Dekkera bruxellensis; Gene expression; Glucose fermentation; Promoters; TEF1