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Abstract
Alkaline extraction followed by acid precipitation were employed to extract rapeseed protein and, Alcalase 2.4 L was used to obtain rapeseed protein hydrolysates. Three groups of rapeseed protein hydrolysates were obtained by purifying with membrane ultrafiltration and a Sephacryl S-100HR gel column. The antioxidant activities were then determined. Group 3 had the best antioxidant activities according to the oxygen radical absorbance capacity, peroxyl radical-scavenging capacity, and cellular antioxidant activity assays, with the following antioxidant values: an oxygen radical absorbance capacity value of 1610 ± 113 µmol TE/(g sample), a peroxyl radical-scavenging capacity value of 622 ± 30 mg VC/(100 g sample), and a cellular antioxidant activity value of 25 ± 2 µmol QE/(g sample) and corresponding EC50 value of 58 ± 3 µg/mL. Six peaks of group 3 were collected and well separated by reversed phase–high-performance liquid chromatography. Peak 5 were identified to exhibit a higher antioxidant activity, the amino acid sequence of which was found to be Trp-Ile (Leu)-Tyr, as determined by liquid chromatography–mass spectrometry.
FUNDING
This work has been supported by the the National Natural Science Foundation of China (No. 31571766), Natural Science Foundation of the Higher Education Institutions of Jiangsu Province, China (14KJB550004), Natural Science Foundation of Jiangsu Province, China (BK20141485), the prospective Industry-Academy-Research cooperation projects of the Jiangsu province China (BY2015010-01), National High Technology Research and Development Program of China (863 Program) (2013AA102207-2), National agricultural achievements transformation projects, China (2014GB2C100318), the National Science and Technology Pillar Program during the Twelfth Five-year Plan Period (No.2014BAD04B03), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).