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Alpha-ketoglutarate enhances freeze–thaw tolerance and prevents carbohydrate-induced cell death of the yeast Saccharomyces cerevisiae

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Abstract

Stress resistance and fermentative capability are important quality characteristics of baker’s yeast. In the present study, we examined protective effects of exogenous alpha-ketoglutarate (AKG), an intermediate of the tricarboxylic acid cycle and amino acid metabolism, against freeze–thaw and carbohydrate-induced stresses in the yeast Saccharomyces cerevisiae. Growth on AKG-supplemented medium prevented a loss of viability and improved fermentative capacity of yeast cells after freeze–thaw treatment. The cells grown in the presence of AKG had higher levels of amino acids (e.g., proline), higher metabolic activity and total antioxidant capacity, and higher activities of catalase, NADP-dependent glutamate dehydrogenase and glutamine synthase compared to control ones. Both synthesis of amino acids and enhancement of antioxidant system capacity could be involved in AKG-improved freeze–thaw tolerance in S. cerevisiae. Cell viability dramatically decreased under incubation of stationary-phase yeast cells in 2% glucose or fructose solutions (in the absence of the other nutrients) as compared with incubation in distilled water or in 10 mM AKG solution. The decrease in cell viability was accompanied by acidification of the medium, and decrease in cellular respiration, aconitase activity, and levels of total protein and free amino acids. The supplementation with 10 mM AKG effectively prevented carbohydrate-induced yeast death. Protective mechanisms of AKG could be associated with the intensification of respiration and prevention of decreasing protein level as well as with direct antioxidant AKG action.

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Abbreviations

AKG:

Alpha-ketoglutarate

CFU:

Colony-forming unit

DC:

α-dicarbonyl compounds

GDH:

Glutamate dehydrogenase

GR:

Glutathione reductase

GS:

Glutamine synthase

G6PDH:

Glucose-6-phosphate dehydrogenase

KPi:

Potassium phosphate buffer

ROS:

Reactive oxygen species

TAC:

Total antioxidant capacity

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Acknowledgements

We are grateful to Dr. Youshiharu Inoue (Kyoto University, Japan) for the providing S. cerevisiae strain. We express our sincere gratitude to Dr. Dmytro Gospodaryov for polarography measurements and critical reading of the manuscript and Dr. Meet R. Zandawala (Stockholm University) for English correction of the article. The work was partially supported by the Ministry of Education and Science of Ukraine (#0115U002304) to Volodymyr I. Lushchak. Two anonymous reviewers are acknowledged for their highly professional and helpful recommendations for improving the paper.

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Authors and Affiliations

  1. Department of Biochemistry and Biotechnology, Vasyl Stefanyk Precarpathian National University, 57 Shevchenko Str, Ivano-Frankivsk, 76018, Ukraine

    Maria M. Bayliak, Olha V. Hrynkiv, Roksolana V. Knyhynytska & Volodymyr I. Lushchak

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  1. Maria M. Bayliak
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  2. Olha V. Hrynkiv
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  3. Roksolana V. Knyhynytska
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  4. Volodymyr I. Lushchak
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Correspondence to Maria M. Bayliak.

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Communicated by Olaf Kniemeyer.

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Bayliak, M.M., Hrynkiv, O.V., Knyhynytska, R.V. et al. Alpha-ketoglutarate enhances freeze–thaw tolerance and prevents carbohydrate-induced cell death of the yeast Saccharomyces cerevisiae . Arch Microbiol 200, 33–46 (2018). https://doi.org/10.1007/s00203-017-1423-9

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