Compositional analysis of proteins following hydrolysis by immobilized proteases.
Pronase, proteinase K, carboxypeptidases A and B, aminopeptidase M, intestinal mucosa exopeptidases and prolidase, immobilized to derivatized controlled-pore glass beads, were used in a study of total enzymic hydrolysis of proteins. The combined use of immobilized enzymatic and acid hydrolysis, for assessment of protein quality, will give a more accurate chemical score than that afforded by acid hydrolysis alone. Amino acid analysis of enzymic hydrolysates of native protein substrates (beta-lactoglobulin and insulin) yielded 92% of the theoretical values and 103% of the values observed for standard acid hydrolysates. These results suggest that using a combination of immobilized proteases in concert gives essentially total hydrolysis of protein substrates in a time period (18-24 h) comparable to conventional acid hydrolysis methods.
D000596:Amino Acids / Q000032:analysis; D004800:Enzymes, Immobilized; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007328:Insulin / Q000032:analysis; D007700:Kinetics; D007782:Lactoglobulins / Q000032:analysis; D010447:Peptide Hydrolases; D011506:Proteins / Q000032:analysis*; D013305:Streptomyces griseus / Q000201:enzymology; D013696:Temperature; D009764:o-Phthalaldehyde
D000596:Amino Acids; D004800:Enzymes, Immobilized; D007328:Insulin; D007782:Lactoglobulins; D011506:Proteins; D009764:o-Phthalaldehyde; D010447:Peptide Hydrolases