Original scientific paper

https://doi.org/10.17113/ftb.57.01.19.5685

Novel Approach in the Construction of Bioethanol-Producing Saccharomyces cerevisiae Hybrids

Anamarija Štafa orcid.org/0000-0001-7534-2900 ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biology and Microbial Genetics, Kršnjavoga 25, 10000 Zagreb, Croatia
Bojan Žunar orcid.org/0000-0002-6855-7222 ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biology and Microbial Genetics, Kršnjavoga 25, 10000 Zagreb, Croatia
Andrea Pranklin ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biology and Microbial Genetics, Kršnjavoga 25, 10000 Zagreb, Croatia
Antonio Zandona orcid.org/0000-0002-3937-3520 ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biology and Microbial Genetics, Kršnjavoga 25, 10000 Zagreb, Croatia
Marina Svetec Miklenić ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biology and Microbial Genetics, Kršnjavoga 25, 10000 Zagreb, Croatia
Božidar Šantek orcid.org/0000-0001-9583-534X ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Kačićeva 28, 10000 Zagreb, Croatia
Ivan Krešimir Svetec orcid.org/0000-0002-0469-2652 ; University of Zagreb, Faculty of Food Technology and Biotechnology, Department of Biochemical Engineering, Laboratory for Biology and Microbial Genetics, Kršnjavoga 25, 10000 Zagreb, Croatia

Abstract

Bioethanol production from lignocellulosic hydrolysates requires a producer strain that tolerates both the presence of growth and fermentation inhibitors and high ethanol concentrations. Therefore, we constructed heterozygous intraspecies hybrid diploids of Saccharomyces cerevisiae by crossing two natural S. cerevisiae isolates, YIIc17_E5 and UWOPS87-2421, a good ethanol producer found in wine and a strain from the flower of the cactus Opuntia megacantha resistant to inhibitors found in lignocellulosic hydrolysates, respectively. Hybrids grew faster than parental strains in the absence and in the presence of acetic and levulinic acids and 2-furaldehyde, inhibitors frequently found in lignocellulosic hydrolysates, and the overexpression of YAP1 gene increased their survival. Furthermore, although originating from the same parental strains, hybrids displayed different fermentative potential in a CO2 production test, suggesting genetic variability that could be used for further selection of desirable traits. Therefore, our results suggest that the construction of intraspecies hybrids coupled with the use of genetic engineering techniques is a promising approach for improvement or development of new biotechnologically relevant strains of S. cerevisiae. Moreover, it was found that the success of gene targeting (gene targeting fidelity) in natural S. cerevisiae isolates (YIIc17_E5α and UWOPS87-2421α) was strikingly lower than in laboratory strains and the most frequent off-targeting event was targeted chromosome duplication.

Keywords

yeast Saccharomyces cerevisiae; intraspecies hybrids; lignocellulosic hydrolysates; growth and fermentation inhibitors; gene targeting

Hrčak ID:

218873

URI

https://hrcak.srce.hr/218873

Publication date:

29.3.2019.

Article data in other languages: croatian

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