Publication

Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption

Reznicek, O., Facey, S. J., de Waal, P. P., Teunissen, A. W. R. H., de Bont, J. A. M., Nijland, J. G., Driessen, A. J. M. & Hauer, B., 15-Apr-2015, In : Journal of Applied Microbiology. 119, p. 99-111 12 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Reznicek, O., Facey, S. J., de Waal, P. P., Teunissen, A. W. R. H., de Bont, J. A. M., Nijland, J. G., ... Hauer, B. (2015). Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption. Journal of Applied Microbiology, 119, 99-111. https://doi.org/10.1111/jam.12825

Author

Reznicek, Ondrej ; Facey, Sandra J ; de Waal, Paul P ; Teunissen, Aloys W R H ; de Bont, Jan A M ; Nijland, J.G. ; Driessen, A.J.M. ; Hauer, Bernhard. / Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption. In: Journal of Applied Microbiology. 2015 ; Vol. 119. pp. 99-111.

Harvard

Reznicek, O, Facey, SJ, de Waal, PP, Teunissen, AWRH, de Bont, JAM, Nijland, JG, Driessen, AJM & Hauer, B 2015, 'Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption', Journal of Applied Microbiology, vol. 119, pp. 99-111. https://doi.org/10.1111/jam.12825

Standard

Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption. / Reznicek, Ondrej; Facey, Sandra J; de Waal, Paul P; Teunissen, Aloys W R H; de Bont, Jan A M; Nijland, J.G.; Driessen, A.J.M.; Hauer, Bernhard.

In: Journal of Applied Microbiology, Vol. 119, 15.04.2015, p. 99-111.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Reznicek O, Facey SJ, de Waal PP, Teunissen AWRH, de Bont JAM, Nijland JG et al. Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption. Journal of Applied Microbiology. 2015 Apr 15;119:99-111. https://doi.org/10.1111/jam.12825


BibTeX

@article{71075aec0cd346e786156cdd2f7abeaa,
title = "Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption",
abstract = "AIMS: S. cerevisiae does not express any xylose-specific transporters. To enhance the xylose uptake of S. cerevisiae, directed evolution of the Gal2 transporter was performed.METHODS AND RESULTS: Three rounds of error-prone PCR were used to generate mutants with improved xylose transport characteristics. After developing a fast and reliable high-throughput screening assay based on flow cytometry, eight mutants were obtained showing an improved uptake of xylose compared to wild-type GAL2 out of 41,200 single yeast cells. Gal2 variant 2.1 harboring five amino acid substitutions showed an increased affinity towards xylose with a faster overall sugar metabolism of glucose and xylose. Another Gal2 variant 3.1 carrying an additional amino acid substitution revealed an impaired growth on glucose but not on xylose.CONCLUSIONS: Random mutagenesis of the S. cerevisiae Gal2 led to an increased xylose uptake capacity and decreased glucose affinity, allowing improved co-consumption.SIGNIFICANCE AND IMPACT OF THE STUDY: Random mutagenesis is a powerful tool to evolve sugar transporters like Gal2 towards co-consumption of new substrates. Using a high-throughput screening system based on flow through cytometry various mutants were identified with improved xylose transport characteristics. The Gal2 variants in this work are a promising starting point for further engineering to improve xylose uptake from mixed sugars in biomass. This article is protected by copyright. All rights reserved.",
author = "Ondrej Reznicek and Facey, {Sandra J} and {de Waal}, {Paul P} and Teunissen, {Aloys W R H} and {de Bont}, {Jan A M} and J.G. Nijland and A.J.M. Driessen and Bernhard Hauer",
year = "2015",
month = "4",
day = "15",
doi = "10.1111/jam.12825",
language = "English",
volume = "119",
pages = "99--111",
journal = "Journal of Applied Microbiology",
issn = "1364-5072",
publisher = "Wiley",

}

RIS

TY - JOUR

T1 - Improved xylose uptake in Saccharomyces cerevisiae due to directed evolution of galactose permease Gal2 for sugar co-consumption

AU - Reznicek, Ondrej

AU - Facey, Sandra J

AU - de Waal, Paul P

AU - Teunissen, Aloys W R H

AU - de Bont, Jan A M

AU - Nijland, J.G.

AU - Driessen, A.J.M.

AU - Hauer, Bernhard

PY - 2015/4/15

Y1 - 2015/4/15

N2 - AIMS: S. cerevisiae does not express any xylose-specific transporters. To enhance the xylose uptake of S. cerevisiae, directed evolution of the Gal2 transporter was performed.METHODS AND RESULTS: Three rounds of error-prone PCR were used to generate mutants with improved xylose transport characteristics. After developing a fast and reliable high-throughput screening assay based on flow cytometry, eight mutants were obtained showing an improved uptake of xylose compared to wild-type GAL2 out of 41,200 single yeast cells. Gal2 variant 2.1 harboring five amino acid substitutions showed an increased affinity towards xylose with a faster overall sugar metabolism of glucose and xylose. Another Gal2 variant 3.1 carrying an additional amino acid substitution revealed an impaired growth on glucose but not on xylose.CONCLUSIONS: Random mutagenesis of the S. cerevisiae Gal2 led to an increased xylose uptake capacity and decreased glucose affinity, allowing improved co-consumption.SIGNIFICANCE AND IMPACT OF THE STUDY: Random mutagenesis is a powerful tool to evolve sugar transporters like Gal2 towards co-consumption of new substrates. Using a high-throughput screening system based on flow through cytometry various mutants were identified with improved xylose transport characteristics. The Gal2 variants in this work are a promising starting point for further engineering to improve xylose uptake from mixed sugars in biomass. This article is protected by copyright. All rights reserved.

AB - AIMS: S. cerevisiae does not express any xylose-specific transporters. To enhance the xylose uptake of S. cerevisiae, directed evolution of the Gal2 transporter was performed.METHODS AND RESULTS: Three rounds of error-prone PCR were used to generate mutants with improved xylose transport characteristics. After developing a fast and reliable high-throughput screening assay based on flow cytometry, eight mutants were obtained showing an improved uptake of xylose compared to wild-type GAL2 out of 41,200 single yeast cells. Gal2 variant 2.1 harboring five amino acid substitutions showed an increased affinity towards xylose with a faster overall sugar metabolism of glucose and xylose. Another Gal2 variant 3.1 carrying an additional amino acid substitution revealed an impaired growth on glucose but not on xylose.CONCLUSIONS: Random mutagenesis of the S. cerevisiae Gal2 led to an increased xylose uptake capacity and decreased glucose affinity, allowing improved co-consumption.SIGNIFICANCE AND IMPACT OF THE STUDY: Random mutagenesis is a powerful tool to evolve sugar transporters like Gal2 towards co-consumption of new substrates. Using a high-throughput screening system based on flow through cytometry various mutants were identified with improved xylose transport characteristics. The Gal2 variants in this work are a promising starting point for further engineering to improve xylose uptake from mixed sugars in biomass. This article is protected by copyright. All rights reserved.

U2 - 10.1111/jam.12825

DO - 10.1111/jam.12825

M3 - Article

VL - 119

SP - 99

EP - 111

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

ER -

ID: 19269071