Re-evaluation of glycerol utilization in Saccharomyces cerevisiae: characterization of an isolate that grows on glycerol without supporting supplements
Open Access
- 8 November 2013
- journal article
- Published by Springer Science and Business Media LLC in Biotechnology for Biofuels
- Vol. 6 (1), 157
- https://doi.org/10.1186/1754-6834-6-157
Abstract
Glycerol has attracted attention as a carbon source for microbial production processes due to the large amounts of crude glycerol waste resulting from biodiesel production. The current knowledge about the genetics and physiology of glycerol uptake and catabolism in the versatile industrial biotechnology production host Saccharomyces cerevisiae has been mainly based on auxotrophic laboratory strains, and carried out in the presence of growth-supporting supplements such as amino acids and nucleic bases. The latter may have resulted in ambiguous conclusions concerning glycerol growth in this species. The purpose of this study was to re-evaluate growth of S. cerevisiae in synthetic glycerol medium without the addition of supplements. Initial experiments showed that prototrophic versions of the laboratory strains CEN.PK, W303, and S288c did not exhibit any growth in synthetic glycerol medium without supporting supplements. However, a screening of 52 S. cerevisiae isolates for growth in the same medium revealed a high intraspecies diversity. Within this group significant variation with respect to the lag phase and maximum specific growth rate was observed. A haploid segregant of one good glycerol grower (CBS 6412-13A) was selected for detailed analysis. Single deletions of the genes encoding for the glycerol/H+ symporter (STL1), the glycerol kinase (GUT1), and the mitochondrial FAD+-dependent glycerol 3-phosphate dehydrogenase (GUT2) abolished glycerol growth in this strain, implying that it uses the same glycerol utilization pathway as previously identified in auxotrophic laboratory strains. Segregant analysis of a cross between CBS 6412-13A and CEN.PK113-1A revealed that the glycerol growth phenotype is a quantitative trait. Genetic linkage and reciprocal hemizygosity analysis demonstrated that GUT1 CBS 6412-13A is one of the multiple genetic loci contributing to the glycerol growth phenotype. The S. cerevisiae intraspecies diversity with regard to glycerol growth is a valuable starting point to identify the genetic and molecular basis of this phenotype. This knowledge can be applied for further rational strain improvement with the goal of using glycerol as a carbon source in industrial biotechnology processes based on S. cerevisiae as a production organism.Keywords
This publication has 50 references indexed in Scilit:
- Engineering of the glycerol decomposition pathway and cofactor regulation in an industrial yeast improves ethanol productionJournal of Industrial Microbiology & Biotechnology, 2013
- Identification of novel causative genes determining the complex trait of high ethanol tolerance in yeast using pooled-segregant whole-genome sequence analysisGenome Research, 2012
- Eukaryote-to-eukaryote gene transfer events revealed by the genome sequence of the wine yeast Saccharomyces cerevisiae EC1118Proceedings of the National Academy of Sciences of the United States of America, 2009
- Population genomics of domestic and wild yeastsNature, 2009
- Comprehensive polymorphism survey elucidates population structure of Saccharomyces cerevisiaeNature, 2009
- Progress in Metabolic Engineering of Saccharomyces cerevisiaeMicrobiology and Molecular Biology Reviews, 2008
- Transcriptome profiling of Saccharomyces cerevisiae during a transition from fermentative to glycerol-based respiratory growth reveals extensive metabolic and structural remodelingMolecular Genetics and Genomics, 2006
- Dissecting the architecture of a quantitative trait locus in yeastNature, 2002
- A second set of loxP marker cassettes for Cre-mediated multiple gene knockouts in budding yeastNucleic Acids Research, 2002
- Studies on the transformation of intact yeast cells by the LiAc/SS‐DNA/PEG procedureYeast, 1995