Ffz1, a new transporter specific for fructose from Zygosaccharomyces bailii.
The basis of fructophily in the yeast Zygosaccharomyces bailii has been shown to reside in the performance of transport systems for hexoses. In this study, a gene encoding a fructose-specific transporter was characterized. The strategy involved the functional complementation of a Saccharomyces cerevisiae strain that does not take up hexoses (hxt-null strain). This strain was transformed with a genomic library of Z. bailii. One transformant capable of growing on fructose, but not on glucose, was obtained. This transformant did not transport d-[(14)C]glucose, and the kinetic parameters for d-[(14)C]fructose were V(max)=3.3 mmol h(-1) g(-1) and K(m)=80.4 mM. As in the original strain of Z. bailii, fructose uptake was not inhibited by the presence of other hexoses or uranyl. The plasmid responsible for the observed phenotype was found to carry an ORF encoding a 616 amino acid protein with the characteristics of a membrane transporter, which was designated FFZ1 (fructose facilitator Zygosaccharomyces). The impairment in function observed in an S. cerevisiae transformant expressing a truncated Ffz1 protein lacking 67 amino acids at the C-terminus suggests an important role for this terminal part in the proper structure of the transporter.
Pina
Carlos
C
Dep. Botânica e Engenharia Biológica, Instituto Superior de Agronomia, 1349-017 Lisboa, Portugal.
Gonçalves
Paula
P
Centro de Recursos Microbiológicos, SABT, FCT/UNL, 2825-516 Caparica, Portugal.
Prista
Catarina
C
Dep. Botânica e Engenharia Biológica, Instituto Superior de Agronomia, 1349-017 Lisboa, Portugal.
Loureiro-Dias
Maria C
MC
Dep. Botânica e Engenharia Biológica, Instituto Superior de Agronomia, 1349-017 Lisboa, Portugal.
D001692:Biological Transport; D005632:Fructose / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D020068:Zygosaccharomyces / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D005632:Fructose