Abstract
The mutant R33 of the obligatory aerobic yeastRhodotorula glutinis exhibited a defect ind-glucose uptake. Detailed kinetic studies ofd-glucose andd-fructose transport in wild-type and mutant strains provided evidence for the existence in the plasma membrane of a carrier specific for fructose. The transport ofd-fructose in the mutant exhibited saturation kinetics up to 1 mmol/Ld-fructose; at higher concentrations the rate ofd-fructose uptake decreased. In the wild-type strain biphasicd-fructose uptake kinetics were observed; the low-affinity component was not found in the mutant, but the high-affinity transport system persisted. During the exponential phase of growth (ond-glucose) the high-affinityd-fructose system was repressed in the wild-type strain. Mutual competition betweend-fructose andd-glucose as well as the pH dependence of transport of the two hexoses further supported the following conclusion: In the wild-type strain,d-fructose is taken up both by the specific fructose carrier (K T=0.22 mmol/L) and the glucose carrier (K T=9.13 mmol/L). The former does not translocated-glucose, the latter is damaged by the mutation. Finally H+ co-transport and plasma membrane depolarization induced by the onset ofd-fructose transport indicated that the fructose carrier is an H+ symporter.
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References
Alcorn M.E., Griffin C.C.: A kinetic analysis ofd-xylose transport inRhodotorula glutinis Biochim. Biophys. Acta 510, 361–371 (1978).
Bergmeyer H.U., Bernt E., Schmidt F., Stork H.:Methoden der enzymatischen Analyse (H.U. Bergmeyer, Ed.), Vol. 2, pp. 1163–1195. Verlag Chemie, Weinheim 1970.
Eddy A.A.: Proton dependent solute transport.Curr. Topics Membr. Transp.,10, 279–360 (1978).
Harold F.M.: Membranes and energy transduction in bacteria.Curr. Topics Bioenerg. 6, 83–149 (1977).
Hauer R., Höfer M.: Variable H+/substrate stoichiomctries inRhodotorula gracilis are caused by a pH-dependent protonation of the carrier(s).Biochem. J. 208 459–464 (1982).
Höfer M.: Mobile membrane carrier for monosaccharide transport inRhodotorula gracilis.J. Membr. Biol. 3, 73–82 (1970).
Höfer M.: Accumulation of electroneutral and charged carbohydrates by proton cotransport inRhodotorula.Methods Enzymol. 174, 629–653 (1989).
Höfer M., Dahle P.: Glucose repression of inducible enzyme synthesis in the yeastRhodotorula gracilis: effect of cell membrane transport.Eur. J. Biochem. 29, 326–332 (1972).
Höfer M., Künemund A.: Tetraphenylphosphonium ion is a true indicator of negative plasma-membrane potential in the yeastRhodotorula glutinis.Biochem. J. 225, 815–819 (1984).
Höfer M., Misra P.C.: Evidence for a proton/sugar symport in the yeastRhodotorula gracilis (glutinis).Biochem. J. 172, 15–22 (1978).
Janda S., Kotyk A., Tauchová R.: Monosaccharide transport systems in the yeastRhodotorula glutinis.Arch. Microbiol. 111, 151–154 (1976).
Klöppel R., Höfer M.: Transport und Umsatz von Polyalkoholen bei der HefeRhodotorula gracilis (glutinis). II. Induzierbarer Transport und Abbau von Pentitolen.Arch. Microbiol. 107, 335–342 (1976).
Kotyk A., Höfer M.: Uphill transport of sugars in the yeastRhodotorula gracilis.Biochim. Biophys. Acta 102, 410–422 (1965).
Lavi L.E., Hermiller J.B., Griffin C.C.: Carrier-mediated transport ofd-ribose byRhodotorula glutinis.Biochim. Biophys. Acta 648, 1–5 (1981).
Milbradt B., Höfer M.: Isolation and characterization of aRhodotorula glutinis mutant defective in glucose transport.J. Gen. Microbiol. 136, 1961–1965 (1990).
Niemietz C., Hauer R., Höfer M.: Active transport of charged substrates by a proton/sugar cotransport system.Biochem. J. 194, 433–441 (1981).
Niemietz C., Höfer M.: Transport of an anionic substrate by the H+/monosaccharide symport inRhodotorula gracilis: only the protonated form of the carrier is catalytically active.J. Membr. Biol. 80, 235–242 (1984).
Pascual C., Romay C., Sigler K.: Proton extrusion inSaccharomyces cerevisiae mutants in very dilute suspensions.Folia Microbiol. 28, 353–362 (1983).
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Gille, G., Höfer, M. & Sigler, K. Evidence for a specific fructose carrier inRhodotorula glutinis based on kinetic studies with a mutant defective in glucose transport. Folia Microbiol 37, 31–38 (1992). https://doi.org/10.1007/BF02814576
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DOI: https://doi.org/10.1007/BF02814576