Simultaneous quantification of the degree of hydrolysis, protein conversion rate and mean molar weight of peptides released in the course of enzymatic proteolysis - ScienceDirect

Fig. 1. Diagram of the global methodology for the simultaneous quantification of the degree of hydrolysis, protein conversion rate and mean molar weight of peptide released in the course of enzymatic proteolysis based on SE-HPLC analysis. Table 1. Enzyme process conditions (pH and temperature) for hydrolysis. Table 2. Aminograms and characteristics of protein substrates for the quantification of DH value. Fig. 2. Superdex peptide column calibration line drawn with synthesized peptides. The elution conditions were 10 μL injection volume, 0.5 mL min−1 flow rate with water/acetonitrile/TFA mix (69.9/30/0.1, v/v/v) buffer. The calibration equation obtained by linear regression was: MMx = 10−0.12.Vr+4.68 and the determination coefficient (R2) was 0.87. Fig. 3. Size exclusion chromatograms obtained for the hydrolysis of 1% (w/v) BSA (A; A′) and RA (B; B′) with Alcalase 2.4L over 5 or 6 h, at 50 °C, pH 9 and 2 different E/S (A: 1/16; A′: 1/1600; B: 1/13; B′: 1/133). Fig. 4. DH values (), Xp v

Fig. 1. Diagram of the global methodology for the simultaneous quantification of the degree of hydrolysis, protein conversion rate and mean molar weight of peptide released in the course of enzymatic proteolysis based on SE-HPLC analysis. Table 1. Enzyme process conditions (pH and temperature) for hydrolysis. Table 2. Aminograms and characteristics of protein substrates for the quantification of DH value. Fig. 2. Superdex peptide column calibration line drawn with synthesized peptides. The elution conditions were 10 μL injection volume, 0.5 mL min−1 flow rate with water/acetonitrile/TFA mix (6