The protein hydrolysate of purple speckled kidney bean (PSKB) was used as the raw material in this study, and the antioxidant peptide of the PSKB protein hydrolysate was purified using macroporous resin. The XAD-7HP macroporous resin was selected as the best purification material, and the static adsorption-desorption of the purified PSKB antioxidant peptide was optimized. The optimum static adsorption and desorption conditions were as follows: the adsorption capacity reached 11.93 ± 0.11 mg/ml at pH 7 for 24 h, and the desorption capacity was 5.24 ± 0.04 mg/ml with 60% ethanol for 30 min. Under this condition, the amount of antioxidant peptide obtained by adsorption-desorption was the highest. The optimum process conditions were as follows: the appropriate flow rate was 1 ml/min, and the optimal injection volume was 40 ml. The adsorption amount at this time can reach 12.19 ± 0.15 mg/ml. The components with an elution time of 10-30 min were separated using the reversed-phase high-performance liquid chromatography (RP-HPLC) technique to obtain three main components, namely, RP1, RP2, and RP3. The DPPH free radical scavenging ability reached 56.26 ± 0.56, 66.42 ± 0.56, and 78.57 ± 0.56%, respectively, which were 36.65, 46.34 ± 0.56, and 54.39 ± 0.56% higher than those before purification. The amino acid sequences of the three components were identified as Phe-Leu-Val-Asp-Arg-Ile, Phe-Leu-Val-Ala-Pro-Asp-Asp, and Lys-Asp-Arg-Val-Ile-Ser-Glu-Leu.
Keywords: antioxidant peptide; macroporous resin; purification; purple speckled kidney bean protein; separation.
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