Under pressure: evolutionary engineering of yeast strains for improved performance in fuels and chemicals production

Mans, Robert; Daran, Jean‐Marc; Pronk, Jack T.

doi:10.1016/j.copbio.2017.10.011

Cited by 141 publications

(112 citation statements)

References 71 publications

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“…Under this change, an increase of proton efflux around 1.5 times can be expected in the TTY23 strain respect to the WT strain (Madeira et al, 2010). Under this change, an increase of proton efflux around 1.5 times can be expected in the TTY23 strain respect to the WT strain (Madeira et al, 2010).…”

Section: This Maladaptation Correlated With the Upregulation Of The Hmentioning

confidence: 92%

“…Facts rather suggested that S. cerevisiae suppression is because of imbalances in cellular energy. Both activities are primarily performed by the H+ATPase PMA1 and needed for nutrients uptake (Madeira et al, 2010;Meena, Thakur, & Chakrabarti, 2011;Serrano, Kielland-Brandt, & Fink, 1986). ATPm is mainly required for keeping H + gradients and electrical potentials in the yeast plasma membrane.…”

Section: Introductionmentioning

confidence: 99%

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Engineering high‐gravity fermentations for ethanol production at elevated temperature with Saccharomyces cerevisiae

Caspeta

Coronel

et al. 2019

Biotech & Bioengineering

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Thermal damage, high osmolarity, and ethanol toxicity in the yeast Saccharomyces cerevisiae limit titer and productivity in fermentation to produce ethanol. We show that long-term adaptive laboratory evolution at 39.5°C generates thermotolerant yeast strains, which increased ethanol yield and productivity by 10% and 70%, in 2% glucose fermentations. From these strains, which also tolerate elevated-osmolarity, we selected a stable one, namely a strain lacking chromosomal duplications. This strain (TTY23) showed reduced mitochondrial metabolism and high proton efflux, and therefore lower ethanol tolerance. This maladaptation was bolstered by reestablishing proton homeostasis through increasing fermentation pH from 5 to 6 and/or adding potassium to the media. This change allowed the TTY23 strain to produce 1.3-1.6 times more ethanol than the parental strain in fermentations at 40°C with glucose concentrations~300 g/L. Furthermore, ethanol titers and productivities up to 93.1 and 3.87 g·L −1 ·hr −1 were obtained from fermentations with 200 g/L glucose in potassium-containing media at 40°C. Albeit the complexity of cellular responses to heat, ethanol, and high osmolarity, in this study we overcome such limitations by an inverse metabolic engineering approach.ethanol tolerance, high-temperature fermentation, S. cerevisiae, thermotolerance Abbreviations: AcetylCoA, acetyl coenzyme-A; ADH, alcohol dehydrogenase; ADH4, alcohol dehydrogenase (alcohol dehydrogenase isoenzyme type IV); ADP, adenosine-diphosphate; ALD2, aldehyde dehydrogenase (cytoplasmic aldehyde dehydrogenase); ATP, adenosine-triphosphate; ATP_F0, ATP synthase, F1 complex (membrane-embedded portion; protons are transferred through this); ATP_F1, ATP synthase, F1 complex (hydrophilic catalytic portion which is sticking into matrix); ATP-Stalk, ATP synthase stalk (it is considered as part of F1); bc1, cytochrome B (mitochondrial cytochrome bc1 complex); CytC, cytochrome C oxidase; Glucose-P, glucose-6-phosphate; Glyceraldehyde-P, glyceraldehyde-3-phosphate; GND1, 6-phosphogluconate dehydrogenase; HSP30, heat shock protein 30 (negative regulator of the H + -ATPase PMA1); IDP1, isocitrate dehydrogenase, NADP-specific (mitochondrial NADP-specific isocitrate dehydrogenase); KHA1, K/H ion antiporter (putative K + /H + antiporter); MDH1, malate dehydrogenase (mitochondrial malate dehydrogenase); MPC1, mitochondrial pyruvate carrier (mitochondrial inner membrane complex that mediates pyruvate uptake); MPC3, mitochondrial pyruvate carrier (highly conserved subunit of the mitochondrial pyruvate carrier); NADPH, nicotinamide adenine dinucleotide reduced; NDE1, NADH dehydrogenase, external (mitochondrial external NADH dehydrogenase; type II NAD(P)H:quinone oxidoreductase that catalyzes the oxidation of cytosolic NADH); NDE2, NADH dehydrogenase external (mitochondrial external NADH dehydrogenase; catalyzes the oxidation of cytosolic NADH); NDI1, NADH dehydrogenase internal (NADH: ubiquinone oxidoreductase; transfers electrons from NADH to ubiquinone in respiratory chain ...

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Section: This Maladaptation Correlated With the Upregulation Of The Hmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Engineering high‐gravity fermentations for ethanol production at elevated temperature with Saccharomyces cerevisiae

Caspeta

Coronel

et al. 2019

Biotech & Bioengineering

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“…Consequently, the evolved strain obtained in this study is compatible with industrial processes that involve acetic acid‐containing hydrolysates along with other inhibitors such as phenolic compounds, and thus shows great potential as a microbial platform in a lignocellulosic biorefinery. The acquired novel stress tolerance of the evolved strains is not always expressed when the selective pressure is alleviated or changed (Mans, Daran, & Pronk, ; Wright et al, ). However, our evolved strain constitutively maintained its high performance of xylose fermentation in acetic acid‐free medium, highlighting its robustness under various conditions.…”

Section: Discussionmentioning

confidence: 99%

Improved bioconversion of lignocellulosic biomass by Saccharomyces cerevisiae engineered for tolerance to acetic acid

Gong

et al. 2019

GCB Bioenergy

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Lignocellulosic biomass has considerable potential for the production of fuels and chemicals as a promising alternative to conventional fossil fuels. However, the bioconversion of lignocellulosic biomass to desired products must be improved to reach economic viability. One of the main technical hurdles is the presence of inhibitors in biomass hydrolysates, which hampers the bioconversion efficiency by biorefinery microbial platforms such as Saccharomyces cerevisiae in terms of both production yields and rates. In particular, acetic acid, a major inhibitor derived from lignocellulosic biomass, severely restrains the performance of engineered xylose‐utilizing S. cerevisiae strains, resulting in decreased cell growth, xylose utilization rate, and product yield. In this study, the robustness of XUSE, one of the best xylose‐utilizing strains, was improved for the efficient conversion of lignocellulosic biomass into bioethanol under the inhibitory condition of acetic acid stress. Through adaptive laboratory evolution, we successfully developed the evolved strain XUSAE57, which efficiently converted xylose to ethanol with high yields of 0.43–0.50 g ethanol/g xylose even under 2–5 g/L of acetic stress. XUSAE57 not only achieved twofold higher ethanol yields but also improved the xylose utilization rate by more than twofold compared to those of XUSE in the presence of 4 g/L of acetic acid. During fermentation of lignocellulosic hydrolysate, XUSAE57 simultaneously converted glucose and xylose with the highest ethanol yield reported to date (0.49 g ethanol/g sugars). This study demonstrates that the bioconversion of lignocellulosic biomass by an engineered strain could be significantly improved through adaptive laboratory evolution for acetate tolerance, which could help realize the development of an economically feasible lignocellulosic biorefinery to produce fuels and chemicals.

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“…Individual cells in a microbial population randomly accumulate mutations, either as a consequence of natural DNA replication errors or via externally induced mutagenesis mechanisms . Some (beneficial) mutations encode phenotypic changes that allow mutants to grow and divide faster than other cells under specific culture conditions, eventually taking progressive control of the entire population . As the selective pressure is increased, other beneficial mutations can be selected until the desired objective is achieved via gradual increases in fitness .…”

Section: Introductionmentioning

confidence: 99%

Evolutionary Approaches for Engineering Industrially Relevant Phenotypes in Bacterial Cell Factories

Fernández-Cabezón

Cros

Nikel

2019

Biotechnology Journal

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The bio‐based production of added‐value compounds (with applications as pharmaceuticals, biofuels, food ingredients, and building blocks) using bacterial platforms is a well‐established industrial activity. The design and construction of microbial cell factories (MCFs) with robust and stable industrially relevant phenotypes, however, remains one of the biggest challenges of contemporary biotechnology. In this review, traditional and cutting‐edge approaches for optimizing the performance of MCFs for industrial bioprocesses, rooted on the engineering principle of natural evolution (i.e., genetic variation and selection), are discussed. State‐of‐the‐art techniques to manipulate and increase genetic variation in bacterial populations and to construct combinatorial libraries of strains, both globally (i.e., genome level) and locally (i.e., individual genes or pathways, and entire sections and gene clusters of the bacterial genome) are presented. Cutting‐edge screening and selection technologies applied to isolate MCFs displaying enhanced phenotypes are likewise discussed. The review article is closed by presenting future trends in the design and construction of a new generation of MCFs that will contribute to the long‐sought‐after transformation from a petrochemical industry to a veritable sustainable bio‐based industry.

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Under pressure: evolutionary engineering of yeast strains for improved performance in fuels and chemicals production

Cited by 141 publications

References 71 publications

Engineering high‐gravity fermentations for ethanol production at elevated temperature with Saccharomyces cerevisiae

Engineering high‐gravity fermentations for ethanol production at elevated temperature with Saccharomyces cerevisiae

Improved bioconversion of lignocellulosic biomass by Saccharomyces cerevisiae engineered for tolerance to acetic acid

Evolutionary Approaches for Engineering Industrially Relevant Phenotypes in Bacterial Cell Factories

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