Isolation of cobalt hyper-resistant mutants of Saccharomyces cerevisiae by in vivo evolutionary engineering approach

Çakar, Z. Petek; Alkım, Ceren; Turanlı, Burcu; Tokman, Nilgün; Akman, Süleyman; Sarıkaya, Mehmet; Tamerler, Candan; Benbadis, Laurent; François, Jean

doi:10.1016/j.jbiotec.2009.06.024

Cited by 50 publications

(39 citation statements)

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“…In this work, we obtained copper-enriched and copper-tolerant yeasts through a number of generations smaller than reported in similar recent works [13,14,25]. This might be explained on the basis of a different experimental set-up in which relatively few rounds of selection were applied, but with longer cultivation times (72 h) and wider intervals of metal concentration (increases of 0.5 g. L -1 at each round).…”

Section: Discussionmentioning

confidence: 82%

“…In this light, the variability in copper sensitivity between Candida and Saccharomyces cells might depend (at least partly) on a different ability to limit copper uptake and its overload. Such a mechanism has been reported to protect S. cerevisiae cells from copper [26] cadmium [27] and cobalt [14] and points to a central role of the plasma membrane [28-30] and of the cell wall [31] in the onset of tolerance to heavy metals.…”

Section: Discussionmentioning

confidence: 99%

“…This approach is often reported as "evolutionary engineering", a term introduced by Butler and collaborators in 1996 [11], since it uses evolutionary principles based on the selection of random mutants arising in the microbial population either spontaneously or upon mutagenesis and has been applied for improving complex physiological properties whose genetic and physiological basis is not fully understood [12]. For example, microbial cells were recently evolved to improve their resistance towards multiple stresses [13], cobalt [14], iron- and sulfur-compounds [15], alcohols [16]; and to gain the ability to ferment xylose [17] and lactose [18]. …”

Section: Introductionmentioning

confidence: 99%

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Laboratory evolution of copper tolerant yeast strains

et al. 2012

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BackgroundYeast strains endowed with robustness towards copper and/or enriched in intracellular Cu might find application in biotechnology processes, among others in the production of functional foods. Moreover, they can contribute to the study of human diseases related to impairments of copper metabolism. In this study, we investigated the molecular and physiological factors that confer copper tolerance to strains of baker's yeasts.ResultsWe characterized the effects elicited in natural strains of Candida humilis and Saccharomyces cerevisiae by the exposure to copper in the culture broth. We observed that, whereas the growth of Saccharomyces cells was inhibited already at low Cu concentration, C. humilis was naturally robust and tolerated up to 1 g · L-1 CuSO4 in the medium. This resistant strain accumulated over 7 mg of Cu per gram of biomass and escaped severe oxidative stress thanks to high constitutive levels of superoxide dismutase and catalase. Both yeasts were then "evolved" to obtain hyper-resistant cells able to proliferate in high copper medium. While in S. cerevisiae the evolution of robustness towards Cu was paralleled by the increase of antioxidative enzymes, these same activities decreased in evolved hyper-resistant Candida cells. We also characterized in some detail changes in the profile of copper binding proteins, that appeared to be modified by evolution but, again, in a different way in the two yeasts.ConclusionsFollowing evolution, both Candida and Saccharomyces cells were able to proliferate up to 2.5 g · L-1 CuSO4 and to accumulate high amounts of intracellular copper. The comparison of yeasts differing in their robustness, allowed highlighting physiological and molecular determinants of natural and acquired copper tolerance. We observed that different mechanisms contribute to confer metal tolerance: the control of copper uptake, changes in the levels of enzymes involved in oxidative stress response and changes in the copper-binding proteome. However, copper elicits different physiological and molecular reactions in yeasts with different backgrounds.

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Section: Discussionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Laboratory evolution of copper tolerant yeast strains

et al. 2012

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“…However, several more applied studies have started from strains commonly used in the wine, beer, biofuel, and baking industry. The phenotypes targeted include resistance to individual stresses, such as high levels of acetate (Aarnio et al ., 1991) or ethanol (Brown & Oliver, 1982; Dinh et al ., 2008), osmotic stress (Ekberg et al ., 2013), and high concentrations of metal ions such as copper (Adamo et al ., 2012) and cobalt (Cakar et al ., 2009), as well as (improved) utilization of alternative carbon sources such as xylose and arabinose (starting from metabolically engineered strains, see also below; Sonderegger & Sauer, 2003; Wisselink et al ., 2009; Scalcinati et al ., 2012; Demeke et al ., 2013). However, in industrial settings, cells are often faced with a combination of different stresses: during brewing fermentations for example, cells encounter osmotic stress, high levels of ethanol, and nutrient deprivation (Gibson et al ., 2007).…”

Section: Natural and Artificial Diversitymentioning

confidence: 99%

Improving industrial yeast strains: exploiting natural and artificial diversity

Steensels¹,

Snoek²,

Meersman³

et al. 2014

FEMS Microbiol Rev

395

288

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Yeasts have been used for thousands of years to make fermented foods and beverages, such as beer, wine, sake, and bread. However, the choice for a particular yeast strain or species for a specific industrial application is often based on historical, rather than scientific grounds. Moreover, new biotechnological yeast applications, such as the production of second-generation biofuels, confront yeast with environments and challenges that differ from those encountered in traditional food fermentations. Together, this implies that there are interesting opportunities to isolate or generate yeast variants that perform better than the currently used strains. Here, we discuss the different strategies of strain selection and improvement available for both conventional and nonconventional yeasts. Exploiting the existing natural diversity and using techniques such as mutagenesis, protoplast fusion, breeding, genome shuffling and directed evolution to generate artificial diversity, or the use of genetic modification strategies to alter traits in a more targeted way, have led to the selection of superior industrial yeasts. Furthermore, recent technological advances allowed the development of high-throughput techniques, such as ‘global transcription machinery engineering’ (gTME), to induce genetic variation, providing a new source of yeast genetic diversity.

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“…It is particularly useful for obtaining genetically complex microbial phenotypes such as tolerance to inhibitors/toxic compounds or various stress types [39]. Successful results were obtained by our research group, regarding evolutionary engineering of multi-stress resistant [40], cobalt-resistant [41,42], nickel-resistant [43], and ethanol-tolerant [44] S. cerevisiae. Another example for the use of evolutionary engineering against product toxicity involves adaptive evolution for lactic acid tolerance in S. cerevisiae [31].…”

Section: Production Of Bulk Chemicalsmentioning

confidence: 99%

Advances in Metabolic Engineering of Saccharomyces cerevisiae for the Production of Industrially and Clinically Important Chemicals

Turanlı-Yıldız¹,

Hacısalihoğlu²,

Çakar³

2017

Old Yeasts - New Questions

Self Cite

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Sustainable production of chemicals is of increasing importance, due to depletion of petroleum and environmental concerns. In addition to its importance in basic research as a simple, eukaryotic model organism, Saccharomyces cerevisiae has long been exploited in industry because of its physiological properties. And today, the development in genetic engineering toolbox and genome-scale metabolic models of S. cerevisiae has extended its application range to new products and bioprocesses. In addition, evolutionary engineering strategies have been useful in improving cellular properties of S. cerevisiae, such as tolerance to product toxicity and inhibitors. In this chapter, recent metabolic and evolutionary engineering studies that involve S. cerevisiae for the production of bulk chemicals and ine chemicals including lavours and pharmaceuticals are reviewed. It was shown that metabolic engineering particularly allowed the improvement of pharmaceuticals production, which will enable economic and large-scale production of many valuable pharmaceuticals. It is clear that S. cerevisiae will continue to be an important host for future metabolic engineering and metabolic pathway engineering applications to produce a variety of industrially and clinically important chemicals.

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Isolation of cobalt hyper-resistant mutants of Saccharomyces cerevisiae by in vivo evolutionary engineering approach

Cited by 50 publications

References 35 publications

Laboratory evolution of copper tolerant yeast strains

Laboratory evolution of copper tolerant yeast strains

Improving industrial yeast strains: exploiting natural and artificial diversity

Advances in Metabolic Engineering of Saccharomyces cerevisiae for the Production of Industrially and Clinically Important Chemicals

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