Re-evaluation of glycerol utilization in Saccharomyces cerevisiae: characterization of an isolate that grows on glycerol without supporting supplements

Biotechnology for Biofuels

Steve Swinnen¹  Mathias Klein¹  Martina Carrillo¹  Joseph McInnes¹  Huyen Thi Thanh Nguyen¹  Elke Nevoigt¹ 

[1] School of Engineering and Science, Jacobs University Bremen gGmbH, Campus Ring 1, 28759 Bremen, Germany

关键词: GUT2;    GUT1;    STL1;    Glycerol;    Saccharomyces cerevisiae;    Yeast;   

Others  :  1085005
DOI  :  10.1186/1754-6834-6-157

 received in 2013-08-05, accepted in 2013-10-29,  发布年份 2013

【 摘 要 】

Background

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.

Results

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.

Conclusion

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.

【 授权许可】

   
2013 Swinnen et al.; licensee BioMed Central Ltd.

【 图 表 】

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