Transport of lactic acid in Kluyveromyces marxianus: evidence for a monocarboxylate uniport.
Lactic acid-grown cells of a strain of Kluyveromyces marxianus transported D- and L-lactic acid by a saturable mechanism that was partially inducible and subject to glucose repression, with the following kinetic parameters at pH 5.4: Vmax = 1.00 (+/- 0.13) mmol h-1 per g dry weight and Ks = 0.42 (+/- 0.08) mM. Lactic acid transport was competitively inhibited by pyruvic, glycolic, acetic and bromoacetic acids. The latter, a non-metabolizable analogue, was transiently accumulated, the extent depending on the extracellular pH. The pH dependence of the Ks values for undissociated lactic acid and for the lactate anion indicated that the latter was the transported species. Lactate uptake was not accompanied by the simultaneous uptake of protons, potassium ions or sodium ions excluding symport mechanisms. Initial lactic acid uptake led to transient membrane hyperpolarization as measured with a fluorescent dye excluding also an electroneutral anion antiport mechanism. It was concluded that lactate anions use a monocarboxylate uniport and that the counter anion, possibly bicarbonate, uses a separate channel, the coupling being electrical and loose.
Fonseca
A
A
Laboratory of Microbiology, Gulbenkian Institute of Science, Oeiras, Portugal.
Spencer-Martins
I
I
van Uden
N
N
D000085:Acetates / Q000032:analysis / Q000378:metabolism; D001693:Biological Transport, Active; D002264:Carboxylic Acids / Q000378:metabolism*; D003470:Culture Media; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007716:Kluyveromyces / Q000378:metabolism*; D007773:Lactates / Q000378:metabolism*; D019344:Lactic Acid
D000085:Acetates; D002264:Carboxylic Acids; D003470:Culture Media; D007773:Lactates; D019344:Lactic Acid; C016274:bromoacetate; D005947:Glucose