In order to investigate the relationship between dietary amino acids and protein, as well as the activities of intestinal sucrase and leucine aminopeptidase in rats, the effects of an amino acid imbalance on these enzyme activities were studied. The amino acid imbalance was created by adding 8% of an indispensable amino acid mixture lacking threonine to a 6% casein diet supplemented with 0.3% methionine. The food intake and growth of rats fed the imbalanced diet ad libitum were depressed, and the segmental weights of the small intestine and its sucrase activity were clearly lower than those of rats fed the basal diet. The effect of the imbalanced diet under pair-feeding condition on the sucrase activity was similar to that under an ad libitum feeding condition. The food intake and segmental sucrase activity, that is, sucrase activity per length of the small intestine, of rats injected with cortisol (1 mg/day) and fed the imbalanced diet were not depressed, although administration of insulin (1.5 U/day) had no effect on the food intake or segmental sucrase activity. Force-feeding stimulated growth of rats receiving the imbalanced diet, as well as increasing their segmental sucrase activities. The effects of these different conditions on the leucine aminopeptidase activity of rats receiving the imbalanced diet were obscure. These results suggest that changes in segmental sucrase activity might be mediated by stimulating factors in food intake affected by the composition of ingested amino acids and protein together with sucrose in the gastrointestinal lumen.