Characterization of New Polyol/H+ Symporters in Debaryomyces hansenii

Abstract

Debaryomyces hansenii is a halotolerant yeast that produces and assimilates a wide variety of polyols. In this work we evaluate polyol transport in D. hansenii CBS 767, detecting the occurrence of polyol/H(+) (and sugar/H(+)) symporter activity, through the transient extracellular alkalinization of unbuffered starved cell suspensions. From the D. hansenii genome database, we selected nine ORFs encoding putative transporter proteins to clone in a centromeric plasmid with C-terminal GFP tagging and screened for polyol/H(+) symporters by heterologous expression in Saccharomyces cerevisiae. Five distinct D. hansenii polyol/H(+) symporters were identified and characterized, with different specificities and affinities for polyols, namely one glycerol-specific (DhStl1), one D-galactitol-specific (DhSgl1, Symporter galactitol/H(+) 1), one D-(+)-chiro-inositol-specific (DhSyi1, Symporter D-(+)-chiro-inositol/H(+) 1), one for D-sorbitol/D-mannitol/ribitol/D-arabitol/D-galactitol (DhSyl1, Symporter Polyols 1) and another for D-sorbitol/D-mannitol/ribitol/D-arabitol (DhSyl2, Symporter Polyols 2). This work contributed to the annotation of new yeast polyol transporters, including two specific for uncommon substrates as galactitol and D-(+)-chiro-inositol.

DOI: 10.1371/journal.pone.0088180

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@inproceedings{Pereira2014CharacterizationON, title={Characterization of New Polyol/H+ Symporters in Debaryomyces hansenii}, author={Iliana Pereira and Ana Madeira and Catarina Prista and Maria Conceiç{\~a}o Loureiro-Dias and Maria Jos{\'e} Leandro}, booktitle={PloS one}, year={2014} }