Akta pure 25 l1

Hydrolyzed hazelnut proteins were fractionated using AKTA-Pure 25-L1 fast protein liquid chromatography (FPLC) system (GE Healthcare Life Sciences, Sweden). HiTrap Capto Q or Capto DEAE anion exchange columns were utilized for fractionation (GE Healthcare). Hydrolysate samples were injected into the column at a rate of 1 CV.min⁻¹. Salt containing 20 or 100 mmolL^-1 Tris-HCl buffers (0.6 mmolL^-1 or 0.8 mmolL^-1 NaCl, set to pH 8) were used for the elution of the column-bound compounds. The elution was carried out utilizing a linear salt gradient over 25 or 32 CV for papain, bromelain and pepsin hydrolysates vs. trypsin, chymotrypsin and thermolysin hydrolysates, respectively, which were fractionated and numbered sequentially. Prior to the experiments, a variety of other ion exchange (HiTrap Capto Q, DEAE FF, Capto DEAE, Capto-S) and hydrophobic interaction (HiTrap Phenyl FF, Butyl-S FF, Octyl FF) columns were tested in pre-trials. Absorbance (280 nm) was measured by means of a UV detector. In addition, medium pH, conductivity, column and system pressure values were also monitored. The collected fractions were stored at −20 °C until further use.