Large-scale robot-assisted genome shuffling yields industrial Saccharomyces cerevisiae yeasts with increased ethanol tolerance

Biotechnology for Biofuels
Tim SnoekKevin J Verstrepen

Abstract

During the final phases of bioethanol fermentation, yeast cells face high ethanol concentrations. This stress results in slower or arrested fermentations and limits ethanol production. Novel Saccharomyces cerevisiae strains with superior ethanol tolerance may therefore allow increased yield and efficiency. Genome shuffling has emerged as a powerful approach to rapidly enhance complex traits including ethanol tolerance, yet previous efforts have mostly relied on a mutagenized pool of a single strain, which can potentially limit the effectiveness. Here, we explore novel robot-assisted strategies that allow to shuffle the genomes of multiple parental yeasts on an unprecedented scale. Screening of 318 different yeasts for ethanol accumulation, sporulation efficiency, and genetic relatedness yielded eight heterothallic strains that served as parents for genome shuffling. In a first approach, the parental strains were subjected to multiple consecutive rounds of random genome shuffling with different selection methods, yielding several hybrids that showed increased ethanol tolerance. Interestingly, on average, hybrids from the first generation (F1) showed higher ethanol production than hybrids from the third generation (F3). In a seco...Continue Reading

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Citations

Jan 14, 2016·Current Genetics·Tim SnoekKarin Voordeckers
May 18, 2016·Applied Microbiology and Biotechnology·Kristoffer KrogerusBrian Gibson
Nov 7, 2015·PLoS Genetics·Karin VoordeckersKevin J Verstrepen
Dec 13, 2016·ACS Synthetic Biology·Matthew J MaurerAdam P Arkin
Mar 24, 2017·Biotechnology for Biofuels·Marcus S BenyaminDavid M Mackie
Jun 6, 2017·FEMS Yeast Research·B GibsonJ P Sampaio
Aug 24, 2017·FEMS Microbiology Reviews·Maria C DzialoKevin J Verstrepen
Dec 3, 2017·Applied and Environmental Microbiology·Kristoffer KrogerusBrian Gibson
Sep 27, 2015·Applied and Environmental Microbiology·Stijn MertensKevin J Verstrepen
Jul 8, 2015·Applied and Environmental Microbiology·Esther MeersmanKevin J Verstrepen
May 24, 2018·Nature Communications·Zhouqing LuoJunbiao Dai
Jan 1, 2020·Biotechnology for Biofuels·Weiting WangMingxiong He
May 19, 2020·Plant, Cell & Environment·Nafiseh GeshnizjaniWilco Ligterink
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Methods Mentioned

BETA
dissection
PCR

Software Mentioned

ScreenMill
ImageJ

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