Strain-dependent tolerance to acetic acid in Dekkera bruxellensis

Moktaduzzaman, Md.; Galafassi, Silvia; Vigentini, Ileana; Foschino, Roberto; Corte, Laura Della; Cardinali, Gianluigi; Piškur, Jure; Compagno, Concetta

doi:10.1007/s13213-015-1115-0

Cited by 18 publications

(17 citation statements)

References 61 publications

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“…On the other hand, no effect of DSF on cell growth was observed when D. bruxellensis was cultivated in galactose even at MIC dose (Figure b). Galactose metabolism in ammonium‐based medium, as in the case of the present work, is slow and might avoid the overflow to the upper part of the glycolysis, leading to a fully respiratory metabolism and releasing cells from GCR (Moktaduzzaman, Galafassi, Vigentini, et al, ). In addition, glycerol was used as a carbon source that is exclusively metabolized by the mitochondria in a respiratory metabolism.…”

Section: Resultsmentioning

confidence: 72%

First aspects on acetate metabolism in the yeast Dekkera bruxellensis: a few keys for improving ethanol fermentation

Teles

Silva

Mendonça

et al. 2018

Yeast

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Dekkera bruxellensis is continuously changing its status in fermentation processes, ranging from a contaminant or spoiling yeast to a microorganism with potential to produce metabolites of biotechnological interest. In spite of that, several major aspects of its physiology are still poorly understood. As an acetogenic yeast, minimal oxygen concentrations are able to drive glucose assimilation to oxidative metabolism, in order to produce biomass and acetate, with consequent low yield in ethanol. In the present study, we used disulfiram to inhibit acetaldehyde dehydrogenase activity to evaluate the influence of cytosolic acetate on cell metabolism. D. bruxellensis was more tolerant to disulfiram than Saccharomyces cerevisiae and the use of different carbon sources revealed that the former yeast might be able to export acetate (or acetyl-CoA) from mitochondria to cytoplasm. Fermentation assays showed that acetaldehyde dehydrogenase inhibition re-oriented yeast central metabolism to increase ethanol production and decrease biomass formation. However, glucose uptake was reduced, which ultimately represents economical loss to the fermentation process. This might be the major challenge for future metabolic engineering enterprises on this yeast.

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Section: Resultsmentioning

confidence: 72%

First aspects on acetate metabolism in the yeast Dekkera bruxellensis: a few keys for improving ethanol fermentation

Teles

Silva

Mendonça

et al. 2018

Yeast

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“…1); these growth kinetics were reproducible. In contrast, we noticed that the culture of strain CBS 98, which was one of the less-tolerant strains (30), behaved in a less reproducible way, with adaptive phases lasting 70 to 300 h (see Fig. 1 for an example); additionally, some cultures did not grow at all.…”

Section: Resultsmentioning

confidence: 88%

“…In particular, the cultivation in liquid medium under aerobic conditions revealed that also the growth of the most-tolerant strain, CBS 4482, was negatively affected by the presence of 120 mM acetic acid. It caused a reduced growth rate, a reduced specific glucose consumption rate, and a reduced specific ethanol production rate (30). In a new set of experiments, strain CBS 4482 showed an initial adaptive phase that lasted approximately 60 h and a subsequent faster exponential-growth phase (growth rate, 0.06 h Ϫ1 ) during shake flask cultivation in liquid glucose-based medium containing 120 mM acetic acid at pH 4.5 (Fig.…”

Section: Resultsmentioning

confidence: 94%

“…In a previous study, we screened the acetic acid tolerances of 29 strains of D. bruxellensis at concentrations up to 120 mM at pH 4.5 and revealed considerable diversity in acetic acid tolerance among the strains (30). In particular, the cultivation in liquid medium under aerobic conditions revealed that also the growth of the most-tolerant strain, CBS 4482, was negatively affected by the presence of 120 mM acetic acid.…”

Section: Resultsmentioning

confidence: 99%

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Effects of Oxygen Availability on Acetic Acid Tolerance and Intracellular pH in Dekkera bruxellensis

Capusoni

Arioli

Zambelli

et al. 2016

Appl Environ Microbiol

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The yeast Dekkera bruxellensis, associated with wine and beer production, has recently received attention, because its high ethanol and acid tolerance enables it to compete with Saccharomyces cerevisiae in distilleries that produce fuel ethanol. We investigated how different cultivation conditions affect the acetic acid tolerance of D. bruxellensis. We analyzed the ability of two strains (CBS 98 and CBS 4482) exhibiting different degrees of tolerance to grow in the presence of acetic acid under aerobic and oxygen-limited conditions. We found that the concomitant presence of acetic acid and oxygen had a negative effect on D. bruxellensis growth. In contrast, incubation under oxygen-limited conditions resulted in reproducible growth kinetics that exhibited a shorter adaptive phase and higher growth rates than those with cultivation under aerobic conditions. This positive effect was more pronounced in CBS 98, the more-sensitive strain. Cultivation of CBS 98 cells under oxygen-limited conditions improved their ability to restore their intracellular pH upon acetic acid exposure and to reduce the oxidative damage to intracellular macromolecules caused by the presence of acetic acid. This study reveals an important role of oxidative stress in acetic acid tolerance in D. bruxellensis, indicating that reduced oxygen availability can protect against the damage caused by the presence of acetic acid. This aspect is important for optimizing industrial processes performed in the presence of acetic acid. IMPORTANCEThis study reveals an important role of oxidative stress in acetic acid tolerance in D. bruxellensis, indicating that reduced oxygen availability can have a protective role against the damage caused by the presence of acetic acid. This aspect is important for the optimization of industrial processes performed in the presence of acetic acid.A s living microorganisms grow, their metabolism causes considerable changes to their ecological niches: nutrients are sequestered, and a variety of compounds are produced, such as organic acids, ethanol, and others, which can create a hostile environment for other competing microorganisms. Bacteria and yeasts are able to produce large amounts of some organic acids, such as acetic acid. In addition, organisms are constantly faced with different environmental stimuli, stresses, and competition with other organisms, which represent the driving forces leading to the evolution of several traits. The yeast Saccharomyces cerevisiae exhibits a strong fermentative lifestyle due to the Crabtree effect and to its ability to grow at a high rate even under anaerobic conditions (1-3) and low pH (4). In the context of natural evolution, this ability may have helped this organism to consume sugars quickly and to compete with other microorganisms by producing ethanol (5, 6). During yeast evolution, this particular strategy apparently evolved in at least two lineages: the Saccharomyces complex and Dekkera/Brettanomyces (7). S. cerevisiae is used worldwide for baking, producing alcoholic bev...

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“…In this study, a more detailed analysis has been carried out to investigate the short-term ethanol response of both parental and engineered strains. A FTIR-based assay, already employed as a powerful technique for ecotoxicological assessments [28][29][30], was carried out to estimate the type and extent of perturbations induced by ethanol stress on both parental and recombinant yeast strains. Ethanol levels were specifically chosen based on the alcohol concentrations typical of the bioethanol industry [18,22].…”

Section: Ftir Fingerprints Under Ethanol Stressmentioning

confidence: 99%

Delta-Integration of Single Gene Shapes the Whole Metabolomic Short-Term Response to Ethanol of Recombinant Saccharomyces cerevisiae Strains

et al. 2020

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In yeast engineering, metabolic burden is often linked to the reprogramming of resources from regular cellular activities to guarantee recombinant protein(s) production. Therefore, growth parameters can be significantly influenced. Two recombinant strains, previously developed by the multiple δ-integration of a glucoamylase in the industrial Saccharomyces cerevisiae 27P, did not display any detectable metabolic burden. In this study, a Fourier Transform InfraRed Spectroscopy (FTIR)-based assay was employed to investigate the effect of δ-integration on yeast strains' tolerance to the increasing ethanol levels typical of the starch-to-ethanol industry. FTIR fingerprint, indeed, offers a holistic view of the metabolome and is a well-established method to assess the stress response of microorganisms. Cell viability and metabolomic fingerprints have been considered as parameters to detecting any physiological and/or metabolomic perturbations. Quite surprisingly, the three strains did not show any difference in cell viability but metabolomic profiles were significantly altered and different when the strains were incubated both with and without ethanol. A LC/MS untargeted workflow was applied to assess the metabolites and pathways mostly involved in these strain-specific ethanol responses, further confirming the FTIR fingerprinting of the parental and recombinant strains. These results indicated that the multiple δ-integration prompted huge metabolomic changes in response to short-term ethanol exposure, calling for deeper metabolomic and genomic insights to understand how and, to what extent, genetic engineering could affect the yeast metabolome.

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Strain-dependent tolerance to acetic acid in Dekkera bruxellensis

Cited by 18 publications

References 61 publications

First aspects on acetate metabolism in the yeast Dekkera bruxellensis: a few keys for improving ethanol fermentation

First aspects on acetate metabolism in the yeast Dekkera bruxellensis: a few keys for improving ethanol fermentation

Effects of Oxygen Availability on Acetic Acid Tolerance and Intracellular pH in Dekkera bruxellensis

Delta-Integration of Single Gene Shapes the Whole Metabolomic Short-Term Response to Ethanol of Recombinant Saccharomyces cerevisiae Strains

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