Overexpression of ESBP6 improves lactic acid resistance and production in Saccharomyces cerevisiae

Sugiyama, Minetaka; Akase, Shin-pei; Nakanishi, Ryota; Kaneko, Yoshinobu; Harashima, Satoshi

doi:10.1016/j.jbiosc.2016.03.010

Cited by 24 publications

(17 citation statements)

References 38 publications

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“…T. neapolitana fermentation takes place on a wide range of carbohydrate-rich substrates at 80 • C (353.15 K) and pH 6.5. However, the lactic acid concentration in the T. neapolitana fermentation broth is in the lower range (2-10 g/L) compared to lactic acid fermentation (50-150 g/L lactic acid) by homolactic fermentation processes [10,11].…”

Section: Introductionmentioning

confidence: 96%

Adsorption Behaviour of Lactic Acid on Granular Activated Carbon and Anionic Resins: Thermodynamics, Isotherms and Kinetic Studies

et al. 2017

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Solid-liquid extraction (adsorption or ion exchange) is a promising approach for the in situ separation of organic acids from fermentation broths. In this study, a diluted concentration of lactic acid (<10 g/L) separation from a model fermentation broth by granular activated carbon (GAC) as well as weak (Reillex ® 425 or RLX425) and strong (Amberlite ® IRA-400 or AMB400) base anion exchange resins under various operating conditions was experimentally investigated. Thermodynamic analysis showed that the best lactic acid adsorption performances were obtained at a pH below the pK a value of lactic acid (i.e., 3.86) for GAC and RLX425 by physical adsorption mechanism and above the pK a value for the AMB400 resin by an ion exchange mechanism, respectively. The adsorption capacity for GAC (38.2 mg/g) was the highest, followed by AMB400 (31.2 mg/g) and RLX425 (17.2 mg/g). As per the thermodynamic analysis, the lactic acid adsorbed onto GAC and RLX425 through a physical adsorption mechanism, whereas the lactic acid adsorbed onto AMB400 with an ion exchange mechanism. The Langmuir adsorption isotherm model (R 2 > 0.96) and the pseudo-second order kinetic model (R 2~1 ) fitted better to the experimental data than the other models tested. Postulating the conditions for the real fermentation broth (pH: 5.0-6.5 and temperature: 30-80 • C), the resin AMB400 represents an ideal candidate for the extraction of lactic acid during fermentation.

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Section: Introductionmentioning

confidence: 96%

Adsorption Behaviour of Lactic Acid on Granular Activated Carbon and Anionic Resins: Thermodynamics, Isotherms and Kinetic Studies

et al. 2017

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“…Its cellular localization is not clear according to YeastRGB (26), but one high throughput dataset suggested that Esbp6 is located in the cell periphery (27). Furthermore, an overexpression mutant of ESBP6 was shown to confer tolerance to 6% lactic acid (28).…”

Section: Resultsmentioning

confidence: 99%

Elucidating aromatic acid tolerance at low pH in Saccharomyces cerevisiae using adaptive laboratory evolution

Pereira

Mohamed

Radi

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Toxicity from the external presence or internal production of compounds can reduce the growth and viability of microbial cell factories and compromise productivity. Aromatic compounds are generally toxic for microorganisms, which makes their production in microbial hosts challenging. Here we use adaptive laboratory evolution to generate Saccharomyces cerevisiae mutants tolerant to two aromatic acids, coumaric acid and ferulic acid. The evolution experiments were performed at low pH (3.5) to reproduce conditions typical of industrial processes. Mutant strains tolerant to levels of aromatic acids near the solubility limit were then analyzed by whole genome sequencing, which revealed prevalent point mutations in a transcriptional activator (Aro80) that is responsible for regulating the use of aromatic amino acids as the nitrogen source. Among the genes regulated by Aro80, ESBP6 was found to be responsible for increasing tolerance to aromatic acids by exporting them out of the cell. Further examination of the native function of Esbp6 revealed that this transporter can excrete fusel acids (byproducts of aromatic amino acid catabolism) and this role is shared with at least one additional transporter native to S. cerevisiae (Pdr12). Besides conferring tolerance to aromatic acids, ESBP6 overexpression was also shown to significantly improve the secretion in coumaric acid production strains. Overall, we showed that regulating the activity of transporters is a major mechanism to improve tolerance to aromatic acids. These findings can be used to modulate the intracellular concentration of aromatic compounds to optimize the excretion of such products while keeping precursor molecules inside the cell.

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“…Interestingly, another transporter from the MCH family (MCH3 also known as ESBP6) was recently identified in a screen for multicopy suppression of lactic acid toxicity even though MCH3 was not regulated by exposure to lactic acid (Sugiyama et al, 2016). Thus, at least two MCH transporters of budding yeast contribute to stress resistance while their physiological function is still unknown.…”

Section: Discussionmentioning

confidence: 99%

MCH4is a multicopy suppressor of glycine toxicity inSaccharomyces cerevisiae

Melnykov

Elson

2019

Preprint

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Saccharomyces cerevisiae can either import amino acids from the surrounding or synthesize inside the cell, and both processes are tightly regulated. Disruption of such regulation can result in amino acid toxicity to the cell through mechanisms that are poorly understood. In this study we make use of a mutant strain with deregulated general amino acid permease gene whose growth is inhibited by low concentrations of several amino acids. We carry out multicopy suppression screen with several toxic amino acids and identify MCH4 as a gene that suppresses inhibitory effects of glycine. We find that expression of MCH4 is regulated by osmotic shock but not other kinds of stress. These findings are discussed in the context of possible mechanisms of amino acid toxicity.

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Overexpression of ESBP6 improves lactic acid resistance and production in Saccharomyces cerevisiae

Cited by 24 publications

References 38 publications

Adsorption Behaviour of Lactic Acid on Granular Activated Carbon and Anionic Resins: Thermodynamics, Isotherms and Kinetic Studies

Adsorption Behaviour of Lactic Acid on Granular Activated Carbon and Anionic Resins: Thermodynamics, Isotherms and Kinetic Studies

Elucidating aromatic acid tolerance at low pH in Saccharomyces cerevisiae using adaptive laboratory evolution

MCH4is a multicopy suppressor of glycine toxicity inSaccharomyces cerevisiae

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