The present study was designed to identify alkaline phosphatases in non-permeabilized hyphal cells of the fungus Neurospora crassa by staining these enzymatic activities with a modified azo dye coupling method. Our strategy allowed the identification of three nonspecific alkaline phosphatase activities, one of them possibly being a novel putative enzyme, which is not responsive to either Mg2+ or EDTA. Another alkaline phosphatase activity, whose location in the hyphal cell is regulated by phosphate, is stimulated by Mg2+, inhibited by EDTA, and somehow dependent on the expression of the pho-2+-encoded Pi-repressible alkaline phosphatase.
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