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Transport of Glycerol by Pseudomonas aeruginosa

^a Microbiology Department, Oklahoma State University, Stillwater, Oklahoma 74074

ABSTRACT

In Pseudomonas aeruginosa, the transport of glycerol was shown to be genetically controlled and to be dependent on induction by glycerol. Accumulation of ¹⁴C-glycerol was almost completely absent in uninduced cells and in a transport-negative mutant. Kinetic studies with induced cells suggested that glycerol may be transported by two systems with different affinities for glycerol. Osmotically shocked cells did not transport glycerol, and the supernatant fluid from shocked cells contained glycerol-binding activity demonstrable by equilibrium dialysis. The binding protein was not glycerol kinase. Binding activity was absent in shock fluids from the transport-negative mutant and from uninduced cells. The glycerol-binding protein was partially purified by precipitation with ammonium sulfate. Mild heat treatment completely eliminated the binding activity of shock fluid and of the partially purified protein. Sodium azide and N-ethylmaleimide inhibited both transport by whole cells and binding of glycerol by shock fluid. It is concluded that transport of glycerol by P. aeruginosa involves a binding protein responsible for recognition of glycerol and may occur by facilitated diffusion or active transport. A requirement for energy has not been demonstrated.

¹ Present address: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Mass. 02115.