Highly effective detection of β-galactosidase using a hydrogel-based colorimetric assay
Production of color detection test is based on the use of a synthetic substrate specific for this enzyme – ortho – nitrophenyl pyranoside (ONPG) immobilized by entrapment inside crosslinked structure of hydrogel matrix. Presents of this biocatalysts is recorded by the appearance of a yellow color formed as a result of ONPG enzymatic hydrolysis. The test is performed by application of small amount of solution potentially containing β-galactosidase on the surface of transparent hydrogel matrix. Matrix containing colorless ONPG turns from clear to yellow. Appearance of the color indicates the presence of β-galactosidase. The intensity observed visually after a certain period of time may provide general information regarding concentration of enzyme.
The invention was laboratory – tested with use of Eppendorf tubes and laboratory glass plates.