Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization
The genetic stability and metabolic robustness of production strains is one of the key criteria for the production of bio-based products by microbial fermentation on an industrial scale. These criteria were here explored in an industrial ethanol-producer strain of Saccharomyces cerevisiae able to co...
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Frontiers Media S.A.
2024-03-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1357671/full |
| _version_ | 1797244747308335104 |
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| author | Maëlle Duperray Mathéo Delvenne Jean Marie François Jean Marie François Frank Delvigne Jean-Pascal Capp |
| author_facet | Maëlle Duperray Mathéo Delvenne Jean Marie François Jean Marie François Frank Delvigne Jean-Pascal Capp |
| author_sort | Maëlle Duperray |
| collection | DOAJ |
| description | The genetic stability and metabolic robustness of production strains is one of the key criteria for the production of bio-based products by microbial fermentation on an industrial scale. These criteria were here explored in an industrial ethanol-producer strain of Saccharomyces cerevisiae able to co-ferment D-xylose and L-arabinose with glucose through the chromosomal integration of several copies of pivotal genes for the use of these pentose (C5) sugars. Using batch sequential cultures in a controlled bioreactor that mimics long-term fermentation in an industrial setting, this strain was found to exhibit significant fluctuations in D-xylose and L-arabinose consumption as early as the 50th generation and beyond. These fluctuations seem not related to the few low-consumption C5 sugar clones that appeared throughout the sequential batch cultures at a frequency lower than 1.5% and that were due to the reduction in the number of copies of transgenes coding for C5 sugar assimilation enzymes. Also, subpopulations enriched with low or high RAD52 expression, whose expression level was reported to be proportional to homologous recombination rate did not exhibit defect in C5-sugar assimilation, arguing that other mechanisms may be responsible for copy number variation of transgenes. Overall, this work highlighted the existence of genetic and metabolic instabilities in an industrial yeast which, although modest in our conditions, could be more deleterious in harsher industrial conditions, leading to reduced production performance. |
| first_indexed | 2024-04-24T19:15:55Z |
| format | Article |
| id | doaj.art-0b022c6f2a914a64b8e66b964dae9360 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-24T19:15:55Z |
| publishDate | 2024-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-0b022c6f2a914a64b8e66b964dae93602024-03-26T04:58:39ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852024-03-011210.3389/fbioe.2024.13576711357671Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilizationMaëlle Duperray0Mathéo Delvenne1Jean Marie François2Jean Marie François3Frank Delvigne4Jean-Pascal Capp5Toulouse Biotechnology Institute, INSA/University of Toulouse, CNRS, INRAE, Toulouse, FranceTERRA Research and Teaching Centre, Microbial Processes and Interactions (MiPI), Gembloux Agro-Bio Tech, University of Liège, Gembloux, BelgiumToulouse Biotechnology Institute, INSA/University of Toulouse, CNRS, INRAE, Toulouse, FranceToulouse White Biotechnology, INSA, INRAE, CNRS, Toulouse, FranceTERRA Research and Teaching Centre, Microbial Processes and Interactions (MiPI), Gembloux Agro-Bio Tech, University of Liège, Gembloux, BelgiumToulouse Biotechnology Institute, INSA/University of Toulouse, CNRS, INRAE, Toulouse, FranceThe genetic stability and metabolic robustness of production strains is one of the key criteria for the production of bio-based products by microbial fermentation on an industrial scale. These criteria were here explored in an industrial ethanol-producer strain of Saccharomyces cerevisiae able to co-ferment D-xylose and L-arabinose with glucose through the chromosomal integration of several copies of pivotal genes for the use of these pentose (C5) sugars. Using batch sequential cultures in a controlled bioreactor that mimics long-term fermentation in an industrial setting, this strain was found to exhibit significant fluctuations in D-xylose and L-arabinose consumption as early as the 50th generation and beyond. These fluctuations seem not related to the few low-consumption C5 sugar clones that appeared throughout the sequential batch cultures at a frequency lower than 1.5% and that were due to the reduction in the number of copies of transgenes coding for C5 sugar assimilation enzymes. Also, subpopulations enriched with low or high RAD52 expression, whose expression level was reported to be proportional to homologous recombination rate did not exhibit defect in C5-sugar assimilation, arguing that other mechanisms may be responsible for copy number variation of transgenes. Overall, this work highlighted the existence of genetic and metabolic instabilities in an industrial yeast which, although modest in our conditions, could be more deleterious in harsher industrial conditions, leading to reduced production performance.https://www.frontiersin.org/articles/10.3389/fbioe.2024.1357671/fullphenotypic heterogeneitygenetic stabilityindustrial yeast strainhomologous recombinationxylosearabinose |
| spellingShingle | Maëlle Duperray Mathéo Delvenne Jean Marie François Jean Marie François Frank Delvigne Jean-Pascal Capp Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization Frontiers in Bioengineering and Biotechnology phenotypic heterogeneity genetic stability industrial yeast strain homologous recombination xylose arabinose |
| title | Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization |
| title_full | Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization |
| title_fullStr | Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization |
| title_full_unstemmed | Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization |
| title_short | Genomic and metabolic instability during long-term fermentation of an industrial Saccharomyces cerevisiae strain engineered for C5 sugar utilization |
| title_sort | genomic and metabolic instability during long term fermentation of an industrial saccharomyces cerevisiae strain engineered for c5 sugar utilization |
| topic | phenotypic heterogeneity genetic stability industrial yeast strain homologous recombination xylose arabinose |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1357671/full |
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