Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation
Genetic engineering of industrial microorganisms often suffers from undesirable side effects on essential functions. Reverse engineering is an alternative strategy to improve multifactorial traits like low glycerol/high ethanol yield in yeast fermentation. Previous rational engineering of this trait...
| Autores principales: | , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Springer
2013
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/51456 |
| _version_ | 1855514429275242496 |
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| author | Hubmann, Georg Mathe, L. Foulquié Moreno, Maria R. Duitama, Jorge Nevoigt, E Thevelein, JM |
| author_browse | Duitama, Jorge Foulquié Moreno, Maria R. Hubmann, Georg Mathe, L. Nevoigt, E Thevelein, JM |
| author_facet | Hubmann, Georg Mathe, L. Foulquié Moreno, Maria R. Duitama, Jorge Nevoigt, E Thevelein, JM |
| author_sort | Hubmann, Georg |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Genetic engineering of industrial microorganisms often suffers from undesirable side effects on essential functions. Reverse engineering is an alternative strategy to improve multifactorial traits like low glycerol/high ethanol yield in yeast fermentation. Previous rational engineering of this trait always affected essential functions like growth and stress tolerance. We have screened Saccharomyces cerevisiae biodiversity for specific alleles causing lower glycerol/higher ethanol yield, assuming higher compatibility with normal cellular functionality. Previous work identified ssk1 E330N…K356N as causative allele in strain CBS6412, which displayed the lowest glycerol/ethanol ratio.We have now identified a unique segregant, 26B, that shows similar low glycerol/high ethanol production as the superior parent, but lacks the ssk1 E330N…K356N allele. Using segregants from the backcross of 26B with the inferior parent strain, we applied pooled-segregant whole-genome sequence analysis and identified three minor quantitative trait loci (QTLs) linked to low glycerol/high ethanol production. Within these QTLs, we identified three novel alleles of known regulatory and structural genes of glycerol metabolism, smp1 R110Q,P269Q , hot1 P107S,H274Y and gpd1 L164P as causative genes. All three genes separately caused a significant drop in the glycerol/ethanol production ratio, while gpd1 L164P appeared to be epistatically suppressed by other alleles in the superior parent. The order of potency in reducing the glycerol/ethanol ratio of the three alleles was: gpd1 L164P > hot1 P107S,H274Y ≥ smp1 R110Q,P269Q .Our results show that natural yeast strains harbor multiple specific alleles of genes controlling essential functions, that are apparently compatible with survival in the natural environment. These newly identified alleles can be used as gene tools for engineering industrial yeast strains with multiple subtle changes, minimizing the risk of negatively affecting other essential functions. The gene tools act at the transcriptional, regulatory or structural gene level, distributing the impact over multiple targets and thus further minimizing possible side-effects. In addition, the results suggest polygenic analysis of complex traits as a promising new avenue to identify novel components involved in cellular functions, including those important in industrial applications. |
| format | Journal Article |
| id | CGSpace51456 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2013 |
| publishDateRange | 2013 |
| publishDateSort | 2013 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | CGSpace514562024-05-01T08:15:18Z Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation Hubmann, Georg Mathe, L. Foulquié Moreno, Maria R. Duitama, Jorge Nevoigt, E Thevelein, JM saccharomyces cerevisiae glycerol ethanol quantitative trait loci backcrossing glicerol etanol loci de rasgos cuantitativos retrocruzamiento biotechnology Genetic engineering of industrial microorganisms often suffers from undesirable side effects on essential functions. Reverse engineering is an alternative strategy to improve multifactorial traits like low glycerol/high ethanol yield in yeast fermentation. Previous rational engineering of this trait always affected essential functions like growth and stress tolerance. We have screened Saccharomyces cerevisiae biodiversity for specific alleles causing lower glycerol/higher ethanol yield, assuming higher compatibility with normal cellular functionality. Previous work identified ssk1 E330N…K356N as causative allele in strain CBS6412, which displayed the lowest glycerol/ethanol ratio.We have now identified a unique segregant, 26B, that shows similar low glycerol/high ethanol production as the superior parent, but lacks the ssk1 E330N…K356N allele. Using segregants from the backcross of 26B with the inferior parent strain, we applied pooled-segregant whole-genome sequence analysis and identified three minor quantitative trait loci (QTLs) linked to low glycerol/high ethanol production. Within these QTLs, we identified three novel alleles of known regulatory and structural genes of glycerol metabolism, smp1 R110Q,P269Q , hot1 P107S,H274Y and gpd1 L164P as causative genes. All three genes separately caused a significant drop in the glycerol/ethanol production ratio, while gpd1 L164P appeared to be epistatically suppressed by other alleles in the superior parent. The order of potency in reducing the glycerol/ethanol ratio of the three alleles was: gpd1 L164P > hot1 P107S,H274Y ≥ smp1 R110Q,P269Q .Our results show that natural yeast strains harbor multiple specific alleles of genes controlling essential functions, that are apparently compatible with survival in the natural environment. These newly identified alleles can be used as gene tools for engineering industrial yeast strains with multiple subtle changes, minimizing the risk of negatively affecting other essential functions. The gene tools act at the transcriptional, regulatory or structural gene level, distributing the impact over multiple targets and thus further minimizing possible side-effects. In addition, the results suggest polygenic analysis of complex traits as a promising new avenue to identify novel components involved in cellular functions, including those important in industrial applications. 2013-12 2014-11-12T13:42:20Z 2014-11-12T13:42:20Z Journal Article https://hdl.handle.net/10568/51456 en Open Access Springer Hubmann, Georg; Mathe, Lotte; Foulquie-Moreno, Maria R; Duitama, Jorge; Nevoigt, Elke; Thevelein, Johan M. 2013. Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation. Biotechnology for Biofuels. 6: 87. |
| spellingShingle | saccharomyces cerevisiae glycerol ethanol quantitative trait loci backcrossing glicerol etanol loci de rasgos cuantitativos retrocruzamiento biotechnology Hubmann, Georg Mathe, L. Foulquié Moreno, Maria R. Duitama, Jorge Nevoigt, E Thevelein, JM Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation |
| title | Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation |
| title_full | Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation |
| title_fullStr | Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation |
| title_full_unstemmed | Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation |
| title_short | Identification of multiple interacting alleles conferring low glycerol and high ethanol yield in Saccharomyces cerevisiae ethanolic fermentation |
| title_sort | identification of multiple interacting alleles conferring low glycerol and high ethanol yield in saccharomyces cerevisiae ethanolic fermentation |
| topic | saccharomyces cerevisiae glycerol ethanol quantitative trait loci backcrossing glicerol etanol loci de rasgos cuantitativos retrocruzamiento biotechnology |
| url | https://hdl.handle.net/10568/51456 |
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