We studied biochemical characteristics and the response to low salinity at short and long-term after feeding of alkaline phosphatase (AP) activity in hepatopancreas of the osmoregulator crab Neohelice granulata from Mar Chiquita coastal lagoon (Buenos Aires Province, Argentina) (37°32′–37°45′S 57°19′–57°26′W). The hepatopancreas exhibited a levamisole-insensitive and a levamisole-sensitive AP activity with distinct characteristics. Levamisole-insensitive activity was similar within the range of pH 7.4–9.0 and exhibited a Michaelis–Menten kinetics. Levamisole-sensitive AP activity appeared to be maximal at pH 8.5 and appeared to exhibit an allosteric kinetics. In crabs acclimated to 10 psu (hyper-regulation conditions) levamisole-insensitive and levamisole-sensitive AP activity increased (about 16-fold) over time from short term (2–4 h) to long term (120 h) after feeding while no changes occurred in crabs acclimated to 35 psu (osmoconforming conditions). The changes of AP activity along with the higher values at 120 h after feeding in 10 psu compared with those in 35 psu, and the concomitant changes in proteolytic activity, suggest a role of AP in digestive and metabolic adjustments at the biochemical level upon hyper-regulatory conditions.