Fructose-biphosphatase (EC 126.96.36.199) activity was determined in the proximal and distal parts of the small intestinal mucosa of rats 1-30 days of age. Activity was found to increase to a maximum on about the 10th postnatal day and then to decrease. It was always higher in the proximal than in the distal portion of the gut. The enzyme showed a wide range of pH optimum around 7.0 and was inhibited by AMP. In 10-day-old rats activity determined 24 or 48 h after a single injection of cortisone or triiodothyronine was significantly decreased. This effect was no longer found for cortisone in 14-day-old animals. Weaning 18-day-old rats to a high-fat diet for 5 or 7 days delayed but did not prevent the usual decrease in activity seen at weaning.