Partition of hexadecane to the cell surface of Candida tropicalis: Mechanism for the transport of water-insoluble substrates

The partition of hexadecane to the cell surface of Candida tropicalis was measured by incubating heat-inactivated cells with hexadecane-1-¹⁴C on a gyratory shaker. The free hexadecane was separated by centrifuging the cells through a 15% sucrose solution, and the partitioned hexadecane was quantified by scintillation spectrometry of the samples from the resulting cell sediment. Heat-inactivated cells did not take up hexadecane as determined by a membrane filtration technique involving organic solvent washing. The partitioning was a time-dependent process. The velocity increased by increasing the shake rate of te shaker. At 360 rpm and with baffled flasks, saturation of the cell surface with hexadecane was obtained after a 20 min incubation period. The amount of hexadecane partitioned depended on the initial hexadecane-to-cell concentration ratio. At a ratio of 5 μmol/mg cell protein the highest amount of hexadecane partitioned was measured at 2100 μmol/mg cell protein. At ratios higher than 6 μmol/mg cell protein the cells were no longer sedimentable by centrifugation. The partition of hexadecane to the cell surface was affected by removing the surface layer of the cell wall by Pronase treatment and by using detergents in the partition assay. Pronase treatment lowered the amount of hexadecane partitioned as a consequence of the removal of the lipophilic layer of the cell surface. Detergents influence the partition coefficient and also lowered the amount of hexadecane partitioning to the cell surface. At a low shaking intensity (280 rpm, unbaffled flasks), after Pronase treatment, and in the presence of detergents he uptake of hexadecane by the cells was limited by the partitioning.