The basic hydrolysis parameters, i.e. molecular weight of substrate, reaction time, pH, temperature, and enzyme and substrate concentration, were standardized to maximize sugar yield from dextran. Thus, a 74% conversion of 4% dextran to sugar syrup was reached in 12 h using 6 dextranase units g−1 hydrolysed substrate. The high concentrations of sucrose in the reaction medium did not affect the hydrolysis efficiency nor the activity of Penicillium notatum dextranase. Action patterns of dextranase from P. notatum on dextran were investigated. Analysis of the dextran hydrolysis end products indicated that P. notatum dextranase was a typical endo-enzyme, and isomaltose and isomaltotriose were identified as the primary final products of dextran hydrolysis.