Purification and Properties of α-Mannosidase from Vacuolar Membranes of Yeast Saccharomyces Cerevisiae
Vacuoles of the yeast, Saccharomyces cerevisiae, play important roles in storage of amino-acids and other metabolites (Dürr et al., 1979; Hubber-Wälchi et al., 1979). They contain many hydrolases and contribute to digestive activities in the vacuolar compartments (Wiemken et al., 1979).α-mannosidase is one of such hydrolases and another marker enzyme of the vacuolar membranes (Van Der Wilden et al., 1973) than H+-translocating ATPase (Kakinuma et al., 1981). Therefore, it is a suitable marker to study the functions and biogenesis of the vacuoles. We have succeeded in solubilizing and purifying the enzyme to near homogeneity from commercial baker’s yeast and found that the purified enzyme fraction contained two isoforms. In this report, we will describe a method for solubilization and purification of yeast α-mannosidase and some properties of the two isoforms.
KeywordsVacuolar Membrane Vacuolar Compartment DEAE Dextran Crude Cell Lysate Total Membrane Fraction
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