In order to evaluate the role of the alkaline phosphatase in intestinal transport processes, we studied the influence of known modulators of the alkaline phosphatase (polyclonal anti-calf AP antibodies, theophylline and zinc ions) on the absorption rate of glucose, inorganic phosphate and glucose liberated from glucose-6-phosphate into calf duodenal brush border membrane vesicles. Our results allow the following conclusions: First a direct involvement of the AP in the Na+-dependent glucose absorption is unlikely. Indeed, theophylline inhibits strongly the AP activity but rather stimulates the glucose uptake; second the AP is indirectly involved in glucose absorption from glucose-6-phosphate, if its enzymatic hydrolysis is the only source of glucose. In that case the Na+-dependent uptake of glucose was completely suppressed either by phosphatase specific antibodies or by theophylline; third the positive correlation found with calf intestinal BBMV between the inhibition of AP by AP antibodies or by theophylline and the decrease of rate of Na+-dependent Pi uptake rate suggests that the enzyme plays some role in the Pi absorption. It appears from the present study that the AP is probably not a carrier protein itself, but its hydrolytic activity might nevertheless be important for intestinal absorption. After hydrolysis of phosphoric esters the alcohol residues and Pi can be supplied to their specific carriers. Furthermore, the high Pi affinity of the enzyme at physiological pH values, could even favour a transient sequestration of phosphate, which then could be transferred to the Pi carrier.