Activity-Based Protein Profiling of Retaining α-Amylases in Complex Biological Samples

Amylases are key enzymes in the processing of starch many kingdoms life. They important catalysts industrial biotechnology where they applied in, among others, food and production detergents. In man amylases first digestion to glucose arguably also preferred target therapeutic strategies aimed at treatment type 2 diabetes patients through down-tuning assimilation. Efficient sensitive assays that report selectively on retaining amylase activities irrespective nature complexity biomaterial studied great value both finding new effective human inhibitors discovery microbial with potentially advantageous features for biotechnological application. Activity-based protein profiling (ABPP) glycosidases is inherently suited development such an assay format. We here design synthesis 1,6-epi-cyclophellitol-based pseudodisaccharides equipped a suite reporter entities their use ABPP from saliva, murine tissue as well secretomes fungi grown starch. The activity efficiency probes substantiated by extensive biochemical analysis, selectivity over related endoglycosidases validated structural studies.