Preparation of high Fischer ratio oligopeptide by proteolysis of corn gluten meal

Abstract

A method to obtain an oligopeptide with high Fischer ratio is described. Corn gluten meal (CGM) was hydrolysed with Alcalase 2.4L using a two-step hydrolysis. In the first-step hydrolysis, the enzyme reaction conditions for hydrolysing CGM were optimised by using the orthogonal experimental design, while pH = 8.0, temperature = 55°C, enzyme to substrate ratio (3:97, w/w), and the substrate concentration = 5% were identified as the optimum conditions, under which up to 11.62% degree of hydrolysis (DH) could be obtained. The hydrolysate was then fractionated by ultrafiltration using a membrane with the molecular cutoff of over 10 kD at 20 kPa. For the second-step hydrolysis, the filtrate was adjusted to pH 6.0, then papain was added at 50°C and the mixture was maintained for 3 hours. The hydrolysate was obtained after inactivating papain and centrifuging. Then the salt (mainly NaCl) in the hydrolysate was removed with an ion exchange resin at the speed of 8 times bed volume per hour, and aromatic amino acids were removed through absorption by active carbon. By using Sephadex G-25 gel filtration chromatography, a peptide mixture with low molecular weights between 1000 and 1300 was obtained. Finally, tests on amino acid composition and free amino acid concentration of oligopeptide solution showed that the oligopeptide had a high Fischer ratio of 34.71 and the yield of 11.59%.