Replica plate screening method for detecting phosphatase activity in basidiomycetes using 1-Napthyl Phosphate as a chromogenic substrate

Abstract

A chromogenic substrate depended replica plate assay that can be used to screen phosphatase activity in basidiomycetes is described. 1-napthyl phosphate can be incorporated directly into the agar plates used to culture the organisms of interest. The approach offers several advantages over current methodologies including the ability to monitor the development of phosphatase activity with colony development, and the ability to distinguish between activity arising from cell-bound and cell-free enzyme. Phosphatase activity was confrrmed by staining the replica plate with fast blue RR salt. This enzyme probe was successfully used to detect phosphatase producing basidiomycetes.

KEY WORDS: Phosphatase activity, Basidiomycetes, 1-napthyl phosphate, Fast Blue RR