Bovine lactoferrin hydrolysate (BLH) was prepared with pepsin, fortified with Cu²⁺ (Mn²⁺) 0.64 and 1.28 (0.28 and 0.56) mg/g protein, and then assessed for their activity against human gastric cancer BGC-823 cells. BLH and the four fortified BLH products dose- and time-dependently had growth inhibition on the cells in both short- and long-time experiments. These samples at dose level of 25 mg/mL could stop cell-cycle progression at the G0/G1-phase, damage mitochondrial membrane, and induce cell apoptosis. In total, the fortified BLH products had higher activities in the cells than BLH alone. Moreover, higher Cu/Mn fortification level brought higher effects, and Mn was more effective than Cu to increase these effects. In the treated cells, the apoptosis-related proteins such as Bad, Bax, p53, cytochrome c, caspase-3, and caspase-9 were up-regulated, while Bcl-2 was down-regulated. Caspase-3 activation was also evidenced using a caspase-3 inhibitor, z-VAD-fmk. Thus, Cu- and especially Mn-fortification of BLH brought health benefits such as increased anti-cancer activity in the BGC-823 cells via activating the apoptosis-related proteins to induce cell apoptosis.

Two casein plasteins were prepared from the Neutrase-catalyzed plastein reaction of casein hydrolysates in the presence of proline and phenylalanine, respectively, and assessed for their antihypertensive activities at 300 mg/kg body using rat model. After being oral administered, the two plasteins at initial time of 2–6 h resulted in the rats decreased systolic and diastolic blood pressures of 10–23 and 8–19 mmHg, respectively; however, the measured antihypertensive activities showed decreasing trends as oral administrating time prolonged. The two plasteins also decreased serum levels of angiotensin II, angiotensin I converting enzyme, and aldosterone, but had unclear effect on serum angiotensinogen level. The two plasteins showed same antihypertensive activities without statistical difference (p > 0.05), and were more efficient than casein hydrolysates but less effective than captopril (p < 0.05) to exert antihypertensive effect. It is concluded that the plastein reaction can confer casein hydrolysates with enhanced in vivo antihypertensive activities.