The sucrose hydrolysis and the preference of consumption of glucose instead of fructose were investigated for the production of 5-hydroxy-2-hydroxymethyl-γ -pyrone (HHMP) in the presence of Aspergillus flavus IOC 3974 cultivated in liquid Czapeck medium. Standardized 0.5g of pellets were transferred as inoculum into twelve conical flasks of 250 ml containing 100 ml of medium with different sucrose concentration, which was kept at 120 rpm and 28◦C for 16 days without pH adjustment. Aliquots of 500μl of the broth culture were withdrawn at 24 h intervals and analyzed. The major yield of HHMP was 26g l−1 in 120g l−1 of sucrose. At these conditions, A. flavus produced an invertase capable of hydrolyzing 65% of total sucrose concentration in 24h, and an isomerase capable of converting fructose into glucose. In this work, it focused the preference for glucose and, then, of fructose by A. flavus and the strategy used to produce HHMP.