The activities of major digestive hydrolases (proteases, amylase, lipase and esterases) along the intestine were studied in the burbot Lota lota (L.) using different methods of activity expression. The enzyme activities were determined both in the whole gut segments and in the isolated mucosa, and then expressed in terms of tissue mass and protein content in the samples. Further, the cumulative activities of these enzymes in the pyloric caeca were compared with those in the rest of the intestine to estimate the overall contribution of these regions to digestion. The data obtained suggest the essential role of the pyloric caeca in the digestion of the burbot. In addition, the variations in the pH values along the intestine and the changes in the enzyme activities with incubation temperature were examined. The study proved the method of enzyme activity expression to be a key factor influencing the outcome of the experiment.