Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine
In winemaking, malolactic fermentation (MLF), which converts L-malic acid to L-lactic acid, is often applied after the alcoholic fermentation stage to improve the sensory properties of the wine and its microbiological stability. MLF is usually performed by lactic acid bacteria, which, however, are s...
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2024-12-01
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| author | Egor A. Vasyagin Valery N. Urakov Maksim Yu. Shalamitskiy Sofia N. Cherviak Elena V. Ivanova Valentina I. Zagoruyko Alexey V. Beletsky Andrey L. Rakitin Eugenia S. Mardanova Vitaly V. Kushnirov Nikolai V. Ravin Andrey V. Mardanov |
| author_facet | Egor A. Vasyagin Valery N. Urakov Maksim Yu. Shalamitskiy Sofia N. Cherviak Elena V. Ivanova Valentina I. Zagoruyko Alexey V. Beletsky Andrey L. Rakitin Eugenia S. Mardanova Vitaly V. Kushnirov Nikolai V. Ravin Andrey V. Mardanov |
| author_sort | Egor A. Vasyagin |
| collection | DOAJ |
| description | In winemaking, malolactic fermentation (MLF), which converts L-malic acid to L-lactic acid, is often applied after the alcoholic fermentation stage to improve the sensory properties of the wine and its microbiological stability. MLF is usually performed by lactic acid bacteria, which, however, are sensitive to the conditions of alcoholic fermentation. Therefore, the development of wine yeast strains capable of both alcoholic fermentation and MLF is an important task. Using genome editing, we engineered a modified variant of the triploid wine yeast strain <i>Saccharomyces cerevisiae</i> I-328, in which the <i>CAR1</i> arginase gene was replaced by the malate permease gene from <i>Schizosaccharomyces pombe</i> and the malolactic enzyme gene from <i>Oenococcus oeni</i>. Genome-wide transcriptional profiling confirmed the expression of the introduced genes and revealed a limited effect of the modification on global gene expression. Winemaking experiments show that genome editing did not affect fermentation activity and ethanol production, while use of the modified strain resulted in a tenfold reduction in malate content with simultaneous formation of lactate. The resulting wines had a softer and more harmonious taste compared to wine obtained using the parental strain. Inactivation of arginase, which forms urea and L-ornithine through the breakdown of arginine, also resulted in a twofold decrease in the content of urea and the carcinogenic ethyl carbamate in wine. Thus, the new strain with the replacement of the arginase gene with the MLF gene cassette is promising for use in winemaking. |
| format | Article |
| id | doaj-art-e3a4c47dde7d48c0a026290cd9592194 |
| institution | Kabale University |
| issn | 2304-8158 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj-art-e3a4c47dde7d48c0a026290cd95921942025-01-10T13:17:39ZengMDPI AGFoods2304-81582024-12-011415410.3390/foods14010054Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in WineEgor A. Vasyagin0Valery N. Urakov1Maksim Yu. Shalamitskiy2Sofia N. Cherviak3Elena V. Ivanova4Valentina I. Zagoruyko5Alexey V. Beletsky6Andrey L. Rakitin7Eugenia S. Mardanova8Vitaly V. Kushnirov9Nikolai V. Ravin10Andrey V. Mardanov11Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaBach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaAll-Russian National Research Institute of Viticulture and Winemaking “Magarach”, Russian Academy of Sciences, 298600 Yalta, RussiaAll-Russian National Research Institute of Viticulture and Winemaking “Magarach”, Russian Academy of Sciences, 298600 Yalta, RussiaAll-Russian National Research Institute of Viticulture and Winemaking “Magarach”, Russian Academy of Sciences, 298600 Yalta, RussiaAll-Russian National Research Institute of Viticulture and Winemaking “Magarach”, Russian Academy of Sciences, 298600 Yalta, RussiaInstitute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaInstitute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaInstitute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaBach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaInstitute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaInstitute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, RussiaIn winemaking, malolactic fermentation (MLF), which converts L-malic acid to L-lactic acid, is often applied after the alcoholic fermentation stage to improve the sensory properties of the wine and its microbiological stability. MLF is usually performed by lactic acid bacteria, which, however, are sensitive to the conditions of alcoholic fermentation. Therefore, the development of wine yeast strains capable of both alcoholic fermentation and MLF is an important task. Using genome editing, we engineered a modified variant of the triploid wine yeast strain <i>Saccharomyces cerevisiae</i> I-328, in which the <i>CAR1</i> arginase gene was replaced by the malate permease gene from <i>Schizosaccharomyces pombe</i> and the malolactic enzyme gene from <i>Oenococcus oeni</i>. Genome-wide transcriptional profiling confirmed the expression of the introduced genes and revealed a limited effect of the modification on global gene expression. Winemaking experiments show that genome editing did not affect fermentation activity and ethanol production, while use of the modified strain resulted in a tenfold reduction in malate content with simultaneous formation of lactate. The resulting wines had a softer and more harmonious taste compared to wine obtained using the parental strain. Inactivation of arginase, which forms urea and L-ornithine through the breakdown of arginine, also resulted in a twofold decrease in the content of urea and the carcinogenic ethyl carbamate in wine. Thus, the new strain with the replacement of the arginase gene with the MLF gene cassette is promising for use in winemaking.https://www.mdpi.com/2304-8158/14/1/54wine yeastmalolactic fermentationethyl carbamate<i>Saccharomyces cerevisiae</i>CRISPR/Cas9genome editing |
| spellingShingle | Egor A. Vasyagin Valery N. Urakov Maksim Yu. Shalamitskiy Sofia N. Cherviak Elena V. Ivanova Valentina I. Zagoruyko Alexey V. Beletsky Andrey L. Rakitin Eugenia S. Mardanova Vitaly V. Kushnirov Nikolai V. Ravin Andrey V. Mardanov Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine Foods wine yeast malolactic fermentation ethyl carbamate <i>Saccharomyces cerevisiae</i> CRISPR/Cas9 genome editing |
| title | Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine |
| title_full | Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine |
| title_fullStr | Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine |
| title_full_unstemmed | Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine |
| title_short | Development of a Wine Yeast Strain Capable of Malolactic Fermentation and Reducing the Ethyl Carbamate Content in Wine |
| title_sort | development of a wine yeast strain capable of malolactic fermentation and reducing the ethyl carbamate content in wine |
| topic | wine yeast malolactic fermentation ethyl carbamate <i>Saccharomyces cerevisiae</i> CRISPR/Cas9 genome editing |
| url | https://www.mdpi.com/2304-8158/14/1/54 |
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