The objective of the present study was to characterize nutritionally esterase-1 (ES-1). For this purpose, the effects of replacement of dietary carbohydrates by isoenergetic amounts of either fat or protein on ES-1 activities of plasma and small intestine were studied in male rats. Purified diets differing in the amounts of maize starch plus dextrose, casein and various types of fat were used. Plasma and jejunal ES-1 activities were found to be increased with increasing fat intakes. As to the type of fat, increasing plasma ES-1 activities were induced by coconut fat, olive oil, maize oil and medium-chain triacylglycerols, in this order. Maize oil induced higher jejunal ES-1 activities than coconut fat and olive oil, but had similar effects to medium-chain triacylglycerols. Maize oil was more powerful in increasing plasma ES-1 activity than isoenergetic amounts of casein, but with respect to jejunal ES-1 activity these dietary components were equally effective. It is concluded that the amounts of fat and protein in the diet are important determinants of ES-1 activities in plasma and jejunum.