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Glycerol positive promoters for tailored metabolic engineering of the yeast Saccharomyces cerevisiae.

Authors
  • Ho, Ping-Wei1
  • Klein, Mathias1
  • Futschik, Matthias2, 3
  • Nevoigt, Elke1
  • 1 Department of Life Sciences and Chemistry, Jacobs University Bremen gGmbH, Campus Ring 1, 28759 Bremen, Germany. , (Germany)
  • 2 School of Biomedical & Healthcare Sciences, Plymouth University Peninsula Schools of Medicine and Dentistry, Plymouth, Devon, PL4 8AA, UK.
  • 3 Centre of Marine Sciences (CCMAR), Universidade do Algarve, Faro, 8005-139, Portugal. , (Portugal)
Type
Published Article
Journal
FEMS Yeast Research
Publisher
Oxford University Press
Publication Date
Feb 22, 2018
Identifiers
DOI: 10.1093/femsyr/foy019
PMID: 29481685
Source
Medline
License
Unknown

Abstract

Glycerol offers several advantages as a substrate for biotechnological applications. An important step towards using the popular production host Saccharomyces cerevisiae for glycerol-based bioprocesses have been recent studies in which commonly used S. cerevisiae strains were engineered to grow in synthetic medium containing glycerol as the sole carbon source. For metabolic engineering projects of S. cerevisiae growing on glycerol, characterized promoters are missing. In the current study, we used transcriptome analysis and a yECitrine-based fluorescence reporter assay to select and characterize 25 useful promoters. The promoters of the genes ALD4 and ADH2 showed 4.2- and 3-fold higher activities compared to the well-known strong TEF1 promoter. Moreover, the collection contains promoters with graded activities in synthetic glycerol medium and different degrees of glucose repression. To demonstrate the general applicability of the promoter collection, we successfully used a subset of the characterized promoters with graded activities in order to optimize growth on glycerol in an engineered derivative of CEN.PK, in which glycerol catabolism exclusively occurs via a non-native DHA pathway.

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