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Xylose transport studies with xylose-utilizing Saccharomyces cerevisiae strains expressing heterologous and homologous permeases

  • Applied Genetics and Molecular Biotechnology
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Abstract

In the present study, we modified xylose uptake properties of a recombinant xylose-utilizing yeast Saccharomyces cerevisiae by expression of heterologous and homologous permease-encoding genes. In a mutant yeast strain with the main seven hexose transporter genes deleted, and engineered for xylose utilization, we screened an expression cDNA library of the filamentous fungus Trichoderma reesei (Hypocrea jecorina) for enhanced growth on xylose plates. One cDNA clone with significant homology to fungal sugar transporters was obtained, but when the clone was retransformed into the host, it did not support significant growth on xylose. However, during a long liquid culture of the strain carrying the cDNA clone, adaptive mutations apparently occurred in the host, which led to growth on xylose but not on glucose. The new transporter homologue, Trxlt1 thus appears to code for a protein specific for xylose uptake. In addition, xylose-transporting properties of some homologous hexose transporters were studied. All of them, i.e., Hxt1, Hxt2, Hxt4, and Hxt7 were capable of xylose uptake. Their affinities for xylose varied, K m values between 130 and 900 mM were observed. The single-Hxt strains showed a biphasic growth mode on xylose, alike the Trxlt1 harboring strain. The initial, slow growth was followed by a long lag and finally by exponential growth.

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Acknowledgements

We want to acknowledge John Londesborough for his advice on the uptake studies and analysis of the results, for instructive discussions, as well as important comments on the manuscript. Virve Vidgren is thanked for the help in cloning of the hexose transporter genes and Carmen Limon for the help in sequencing of the xylose transporter. Technical assistance of Seija Rissanen, Aila Siltala, Eila Leino, and Titta Manninen is warmly thanked. This work is part of the research program “VTT Industrial Biotechnology” (Academy of Finland; Finnish Centre of Excellence program, 2000–2005, Project no. 64330). The financial support of Technology Agency of Finland (Project no. Tekes 40416/01) is acknowledged.

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  1. Jenita Rauta

    Present address: Laboratory of Cancer Genetics, Institute of Medical Technology, University of Tampere, Biokatu 6-8, FI-33520,, Tampere, Finland

Authors and Affiliations

  1. VTT, Technical Research Centre of Finland, P.O. Box 1000, Espoo, FI-02044, Finland

    Anu Saloheimo, Jenita Rauta, Merja Penttilä & Laura Ruohonen

  2. Institute of Cell Biology, National Academy of Sciences of Ukraine, Drahomanov Str. 14/16, Lviv, 79005, Ukraine

    Oleh V. Stasyk & Andrei A. Sibirny

  3. Department of Metabolic Engineering, University of Rzeszów, Cwiklinskiej 2, 35-601, Rzeszów, Poland

    Andrei A. Sibirny

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  1. Anu Saloheimo
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Correspondence to Laura Ruohonen.

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Saloheimo, A., Rauta, J., Stasyk, O.V. et al. Xylose transport studies with xylose-utilizing Saccharomyces cerevisiae strains expressing heterologous and homologous permeases. Appl Microbiol Biotechnol 74, 1041–1052 (2007). https://doi.org/10.1007/s00253-006-0747-1

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