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A new laboratory evolution approach to select for constitutive acetic acid tolerance in Saccharomyces cerevisiae and identification of causal mutations

Overview of attention for article published in Biotechnology for Biofuels and Bioproducts, August 2016
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  • Good Attention Score compared to outputs of the same age (69th percentile)
  • High Attention Score compared to outputs of the same age and source (83rd percentile)

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Title
A new laboratory evolution approach to select for constitutive acetic acid tolerance in Saccharomyces cerevisiae and identification of causal mutations
Published in
Biotechnology for Biofuels and Bioproducts, August 2016
DOI 10.1186/s13068-016-0583-1
Pubmed ID
Authors

Daniel González-Ramos, Arthur R. Gorter de Vries, Sietske S. Grijseels, Margo C. van Berkum, Steve Swinnen, Marcel van den Broek, Elke Nevoigt, Jean-Marc G. Daran, Jack T. Pronk, Antonius J. A. van Maris

Abstract

Acetic acid, released during hydrolysis of lignocellulosic feedstocks for second generation bioethanol production, inhibits yeast growth and alcoholic fermentation. Yeast biomass generated in a propagation step that precedes ethanol production should therefore express a high and constitutive level of acetic acid tolerance before introduction into lignocellulosic hydrolysates. However, earlier laboratory evolution strategies for increasing acetic acid tolerance of Saccharomyces cerevisiae, based on prolonged cultivation in the presence of acetic acid, selected for inducible rather than constitutive tolerance to this inhibitor. Preadaptation in the presence of acetic acid was shown to strongly increase the fraction of yeast cells that could initiate growth in the presence of this inhibitor. Serial microaerobic batch cultivation, with alternating transfers to fresh medium with and without acetic acid, yielded evolved S. cerevisiae cultures with constitutive acetic acid tolerance. Single-cell lines isolated from five such evolution experiments after 50-55 transfers were selected for further study. An additional constitutively acetic acid tolerant mutant was selected after UV-mutagenesis. All six mutants showed an increased fraction of growing cells upon a transfer from a non-stressed condition to a medium containing acetic acid. Whole-genome sequencing identified six genes that contained (different) mutations in multiple acetic acid-tolerant mutants. Haploid segregation studies and expression of the mutant alleles in the unevolved ancestor strain identified causal mutations for the acquired acetic acid tolerance in four genes (ASG1, ADH3, SKS1 and GIS4). Effects of the mutations in ASG1, ADH3 and SKS1 on acetic acid tolerance were additive. A novel laboratory evolution strategy based on alternating cultivation cycles in the presence and absence of acetic acid conferred a selective advantage to constitutively acetic acid-tolerant mutants and may be applicable for selection of constitutive tolerance to other stressors. Mutations in four genes (ASG1, ADH3, SKS1 and GIS4) were identified as causative for acetic acid tolerance. The laboratory evolution strategy as well as the identified mutations can contribute to improving acetic acid tolerance in industrial yeast strains.

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X Demographics

The data shown below were collected from the profiles of 6 X users who shared this research output. Click here to find out more about how the information was compiled.
Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 177 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Spain 1 <1%
Netherlands 1 <1%
Unknown 175 99%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 41 23%
Student > Master 27 15%
Researcher 24 14%
Student > Bachelor 14 8%
Student > Doctoral Student 10 6%
Other 24 14%
Unknown 37 21%
Readers by discipline Count As %
Agricultural and Biological Sciences 55 31%
Biochemistry, Genetics and Molecular Biology 50 28%
Engineering 11 6%
Medicine and Dentistry 4 2%
Unspecified 3 2%
Other 11 6%
Unknown 43 24%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 5. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 27 December 2017.
All research outputs
#7,047,954
of 25,374,917 outputs
Outputs from Biotechnology for Biofuels and Bioproducts
#455
of 1,578 outputs
Outputs of similar age
#109,554
of 369,194 outputs
Outputs of similar age from Biotechnology for Biofuels and Bioproducts
#8
of 49 outputs
Altmetric has tracked 25,374,917 research outputs across all sources so far. This one has received more attention than most of these and is in the 71st percentile.
So far Altmetric has tracked 1,578 research outputs from this source. They receive a mean Attention Score of 4.9. This one has gotten more attention than average, scoring higher than 70% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 369,194 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 69% of its contemporaries.
We're also able to compare this research output to 49 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 83% of its contemporaries.