Statistical optimization of protein hydrolysis using mixture design: Development of efficient systems for suppression of lipid accumulation in 3T3 L1 adipocytes
Abstract Soy protein isolate (SPI) bovine whey protein (BWP) and egg white protein (EWP) were hydrolyzed with the protease Flavourzyme(tm) 500 L, and the interactions of these substrates for the inhibition of the relative lipid accumulation (RLA) in 3T3 L1 preadipocytes during differentiation was studied using a simplex centroid mixture design. The results indicated that there were synergistic effects for mixtures of non hydrolyzed and hydrolyzed proteins. The hydrolyzed mixture containing BWP (1/2) and EWP (1/2) at 800 ug mL 1 showed increases of up to 220 and 27% in their activities, respectively, compared to the isolated substrates, reaching a maximum RLA suppression of 15.5% The treatment in which the two day postconfluent 3T3 L1 preadipocytes received 1200 ug mL 1 of the mixture containing BWP (1/2) and EWP (1/2) at day zero and every two days afterwards until the end of the experiment at day eight was demonstrated to be more effective, reaching an RLA suppression of 47.9% The results from the fractionation by ultrafiltration indicated that the non fractionated sample of BWP (1/2) and EWP (1/2) was the most active for the anti adipogenic activity and indicated that there is an important contribution of peptide fractions with various molecular sizes in the RLA suppression in 3T3 L1 cells.