Leaf protein containing approximately 50% rubisco (ribulose bisphosphate carboxylase/oxygenase) was obtained from fresh spinach leaf with the use of a simple extraction method. Pepsin and pepsin-pancreatin digests of spinach leaf protein have potent angiotensin-I converting enzyme inhibitory properties with IC(50) values of 56 and 120 microg/mL, respectively. Both digests of leaf protein have antihypertensive effects after oral administration to spontaneously hypertensive rats (SHR) with minimum effective doses of 0.25 and 0.5 g/kg, respectively. The maximum antihypertensive effect for the pepsin digest was observed 4 h after oral administration, while for the pepsin-pancreatin digest, the maximum effect was observed 2 h after oral administration. Undigested spinach leaf protein did not exert any significant antihypertensive effect after oral administration to SHR at doses of 0.5 and 1 g/kg. Obtained results show that the pepsin digest of leaf protein may be useful in treatment of hypertension.