Assessment of ethanol tolerance of Kluyveromyces marxianus CCT 7735 selected by adaptive laboratory evolution

Silveira, Fernando A.; Soares, Dalila Luzia de Oliveira; Bang, Kyung Whan; Balbino, Thércia Rocha; Ferreira, Maurício Alexander de Moura; Diniz, Raphael Hermano Santos; Lima, Lorena Azevedo de; Brandão, Marcelo Mendes; Villas‐Bôas, Silas G.; Silveira, Wendel Batista da

doi:10.1007/s00253-020-10768-9

Cited by 30 publications

(16 citation statements)

References 68 publications

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“…Recently, in the study of da Silveira et al [43], ethanol-tolerant K. marxianus CCT 7735 strains were developed using the Adaptive laboratory evolution (ALE) strategy [44]. Briefly, hundreds of generations of K. marxianus were exposed to 4% (v/v) ethanol, and the trained yeast cells were considered "ethanol tolerant" when a significant increase (>50%) in the specific growth rate was observed.…”

Section: Advanced Techniques In Kluyveromyces Marxianus Strain Improvmentioning

confidence: 99%

Kluyveromyces marxianus: Current State of Omics Studies, Strain Improvement Strategy and Potential Industrial Implementation

2020

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Bioethanol is considered an excellent alternative to fossil fuels, since it importantly contributes to the reduced consumption of crude oil, and to the alleviation of environmental pollution. Up to now, the baker yeast Saccharomyces cerevisiae is the most common eukaryotic microorganism used in ethanol production. The inability of S. cerevisiae to grow on pentoses, however, hinders its effective growth on plant biomass hydrolysates, which contain large amounts of C5 and C12 sugars. The industrial-scale bioprocessing requires high temperature bioreactors, diverse carbon sources, and the high titer production of volatile compounds. These criteria indicate that the search for alternative microbes possessing useful traits that meet the required standards of bioethanol production is necessary. Compared to other yeasts, Kluyveromyces marxianus has several advantages over others, e.g., it could grow on a broad spectrum of substrates (C5, C6 and C12 sugars); tolerate high temperature, toxins, and a wide range of pH values; and produce volatile short-chain ester. K. marxianus also shows a high ethanol production rate at high temperature and is a Crabtree-negative species. These attributes make K. marxianus promising as an industrial host for the biosynthesis of biofuels and other valuable chemicals.

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Section: Advanced Techniques In Kluyveromyces Marxianus Strain Improvmentioning

confidence: 99%

Kluyveromyces marxianus: Current State of Omics Studies, Strain Improvement Strategy and Potential Industrial Implementation

2020

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“…This type of study benefits from the wide inter- and even intra-strain diversity observed, manifested also in genomic differences during the maintenance of microbial cultures in different environmental conditions, and associated with adaptive microevolutionary changes observed even within the descendants of the same strain [ 115 ]. When considering K. marxianus , strains with higher growth rates were obtained under stressful high ethanol environments [ 116 ], within the range of 7–10% ( v / v ) of ethanol [ 117 ]. In a similar way, a recent study and, to the extent of our knowledge, the only report applying this technique to the generation of T. delbrueckii evolved strains, was conducted by Catrielo et al [ 96 ].…”

Section: Genetic Tools Employed Towards T Delbrueckii Improvementmentioning

confidence: 99%

Biotechnological Importance of Torulaspora delbrueckii: From the Obscurity to the Spotlight

Fernandes

Silva-Sousa

Pereira

et al. 2021

JoF

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Torulaspora delbrueckii has attracted interest in recent years, especially due to its biotechnological potential, arising from its flavor- and aroma-enhancing properties when used in wine, beer or bread dough fermentation, as well as from its remarkable resistance to osmotic and freezing stresses. In the present review, genomic, biochemical, and phenotypic features of T. delbrueckii are described, comparing them with other species, particularly with the biotechnologically well-established yeast, Saccharomyces cerevisiae. We conclude about the aspects that make this yeast a promising biotechnological model to be exploited in a wide range of industries, particularly in wine and bakery. A phylogenetic analysis was also performed, using the core proteome of T. delbrueckii, to compare the number of homologous proteins relative to the most closely related species, understanding the phylogenetic placement of this species with robust support. Lastly, the genetic tools available for T. delbrueckii improvement are discussed, focusing on adaptive laboratorial evolution and its potential.

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“…While there are no studies that report the generation of T. delbrueckii strains resistant to ethanol by adaptive evolution, there are some studies that have reported the use this strategy to improve ethanol resistance in S. cerevisiae (Brown and Oliver, 1982;Dinh et al, 2008;Stanley et al, 2010b;Fiedurek et al, 2011;Chen and Xu, 2014) and Kluyveromyces marxianus (Mo et al, 2019;da Silveira et al, 2020). In this sense, da Silveira et al (2020) reported the generation of four ethanol tolerant strains of K. marxianus CCT 7735 by adaptive laboratory evolution.…”

Section: Adaptive Evolutionmentioning

confidence: 99%

Adaptive Laboratory Evolution of Native Torulaspora delbrueckii YCPUC10 With Enhanced Ethanol Resistance and Evaluation in Co-inoculated Fermentation

2020

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Torulaspora delbrueckii is a yeast species typically present in the early stages of the fermentation process. T. delbrueckii positively modifies the aromatic properties of wines. However, its contribution to the final quality of the wine is restricted by its low tolerance to ethanol. T. delbrueckii is capable of fermenting and tolerating an ethanol concentration ranging from 7.4% (v/v) to slightly higher than 9% (v/v). For this reason, it cannot complete fermentation, when alcohol reach levels higher than 12% (v/v), limiting their use in the industry. The objective of this work was to obtain new variants of T. delbrueckii with improved resistance to ethanol through adaptive laboratory evolution. Variants capable of tolerating ethanol levels of 11.5% (v/v) were obtained. These presented improved kinetic parameters, and additionally showed an increase in resistance to SO2 in ethanol compared to the original strain. Co-inoculated fermentations were performed with the original strain (FTd/Sc) and with the evolved strain (FTdF/Sc), in addition to a control fermentation using only Saccharomyces cerevisiae EC1118 (FSc). The results obtained show that FTdF/Sc present higher levels of 2-Ethylhexanol, compared to FTd/Sc and FSc. Furthermore, FTdF/Sc presents higher levels of total alcohols, total aldehydes, total phenolic derivatives, and total sulfur compounds with significant differences with FSc. These results provide a T. delbrueckii YCPUC10-F yeast with higher resistance to ethanol, which can be present throughout the fermentation process and be used in co-inoculated fermentations. This would positively impact the performance of T. delbrueckii by allowing it to be present not only in the early stages of fermentation but to remain until the end of fermentation.

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Assessment of ethanol tolerance of Kluyveromyces marxianus CCT 7735 selected by adaptive laboratory evolution

Cited by 30 publications

References 68 publications

Kluyveromyces marxianus: Current State of Omics Studies, Strain Improvement Strategy and Potential Industrial Implementation

Kluyveromyces marxianus: Current State of Omics Studies, Strain Improvement Strategy and Potential Industrial Implementation

Biotechnological Importance of Torulaspora delbrueckii: From the Obscurity to the Spotlight

Adaptive Laboratory Evolution of Native Torulaspora delbrueckii YCPUC10 With Enhanced Ethanol Resistance and Evaluation in Co-inoculated Fermentation

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