INTRA CELLULAR AND EXTRACELLULAR FORMS OF ALPHA-GLUCOSIDASE FROM ASPERGILLUS-NIGER

An intracellular α-glucosidase (α-glu1) of Aspergillus niger was purified and its properties were compared to those of a secreted α-glucosidase (α-gluE). The estimated molecular weight of α-gluI was 95,000 by gel filtration (α-gluE = 63,000); it is a glycoprotein possessing 29 mol of mannose, 6 mol of glucosamine, and 14 mol of glucose (α-gluE has 5-6 and 2 mol of mannose and glucosamine, respectively). The Km′s of α-glu1 for p-nitrophenyl-α-d-glucopyranoside and maltose were 1.49 and 1.04, respectively, slightly lower than those of α-gluE. In addition, at 65 °C α-gluI enzymatic activity decayed fivefold faster than that of α-gluE, and anti-α-gluE antibody did not recognize α-gluI. While some of these distinctions between the enzymes could be ascribed to conformational differences, the great dissimilarity in molecular weight (approximately 32,000) and lack of reactivity with anti-α-gluE argue against α-gluI being related to α-gluE. The antibody covalently coupled to horseradish peroxidase (Ab-Px) was used as a probe to determine the cellular location of α-gluE by electron microscopic immunocytology. It was found on both sides of the plasma membrane (pm) and in the outer of the two layers of the cell wall. This may mean that α-gluE is synthesized at the inner surface of the pm, is extruded through the pm, becomes associated with the outer layer of the cell wall (perhaps as enzyme-substrate complex), and is eventually released into the growth medium. © 1979.