Isolation of glucanase-containing particles from budding Saccharomyces cerevisiae

In an investigation of the role of glucanases in modifying yeast cell walls at the location of new buds, vesicles derived from the endoplasmic reticulum, which are secreted locally into the cell wall of growing buds and may be involved in the secretion of glucanases, have been isolated.In yeast, exo-beta-1,3-glucanase (EC 3.2.1.6) is present both extra- and intracellularly. Exponentially growing cells contain about 16% of the enzyme activity intracellularly (within the plasmalemma). Most, if not all, of this intracellular glucanase is sedimentable. Of the three classes of subcellular particles that contain glucanase, one is almost completely absent from stationary phase cells and almost absent from cells of the late budding phase of the budding cycle. These particles were isolated from budding cells by combined differential and density gradient centrifugation. They contain exo- and endo-beta-1,3-glucanases, mannan, and protein. The isolate consists mainly of membrane-bounded particles with diameters corresponding to those of the secretory vesicles observed in situ. It is concluded that these particles are identical with the vesicles derived from the endoplasmic reticulum.