Alkaline phosphatase of Pseudomonas aeruginosa: the mechanism of secretion and release of the enzyme from whole cells

Abstract

The release of alkaline phosphatase from whole cells of Pseudomonas aeruginosa as a function of the MgCl2 concentration is proportional to the release of lipopolysaccharide from the cells. Cells grown under conditions where APase is almost completely secreted to the culture filtrate, i.e. growth at pH 7.6, also secrete lipopolysaccharide. Twenty percent sucrose releases a variable quantity of whole cell phosphatase. Localization of this portion of enzyme by biochemical and electron-microscopic techniques showed that it is located on the cell surface exterior to the outer tripartite layer. Phosphatase, which is not released by sucrose, but which is released by MgCl2, is located in the periplasmic space. Phosphatase is located in three areas; the culture filtrate, the outer cell wall surface, and the periplasmic space. The results suggest that A Pase is associated with, and bound to, a cell wall fraction which contains lipopolysaccharide and that the enzyme is "transported" through the outer wall in complex with this fraction. Liberation of the complex from the outer wall may be accomplished by the mechanical shearing forces developed during growth or during the sucrose suspension procedure.