Casein hydrolysate by Aspergillus oryzae protease has been developed to obtain two kinds of antihypertensive peptides, Val-Pro-Pro and Ile-Pro-Pro. In this study, casein miso paste was prepared by adding casein at various concentrations during miso paste fermentation in order to release angiotensin-I converting enzyme (ACE) inhibitory peptides. Increase of peptide concentration and ACE inhibitory activity in casein miso paste was observed and corresponded to the degradation of casein and soy bean proteins during fermentation for 7 days. ACE inhibitory activity of the casein miso paste was higher than that of the general (casein-free) miso paste after fermentation for 7 days. The levels of ACE inhibitory peptides, Val-Pro-Pro and Ile-Pro-Pro, were increased in the casein miso paste during the fermentation. Significant antihypertensive effects of casein miso paste compared to water and the general miso paste (P<0.05 and P<0.05) were confirmed in spontaneously hypertensive rats at a dosage of 1.8 g of the casein miso paste/kg of BW. Taken into account of minimum effective dosages of Val-Pro-Pro and Ile-Pro-Pro in previous clinical studies, these results suggest that casein miso paste might have the potential to control blood pressure in humans. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.