DETERMINATION OF THE ACTIVITY OF AMYLASES AND PROTEASES USING SUBSTRATE-IMMOBILIZED POLYACRYLAMIDE GEL
A fast and cheap method of quantitative determination of amylase and protease activity is proposed. The method involves the use of polyacrylamide gel with immobilized substrate hydrolase. The test solution is placed in the holes of a standard 96-well immunological tablet, kept in contact with the gel surface, after which the gel is painted and photographed. Quantitative determination of enzymatic activity is carried out using a computer program that allows to determine the color of the gel sites corresponding to individual wells. The possibility of using the method for quantitative determination of the activity of amylases and proteases of different biological origin is investigated. The direct dependence of the obtained numerical data on the enzymatic activity in experiments using amylase Bacillus subtilis, trypsin, amylase and protease of Colorado potato beetle larvae was shown. In all cases, except amylase Bacillus subtilis, there is a nonlinearity of the dependence of the numerical data on the enzyme activity.