Study on separation and purification of antioxidant peptide from pigskin collagen protein and antioxidant activity in vitro
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摘要: 采用碱性蛋白酶对猪皮胶原蛋白进行酶解,制备抗氧化肽。为了得到抗氧化活性高且纯度高的抗氧化肽,本实验采用多种体外抗氧化评价体系研究超滤获得的各分子质量段猪皮胶原蛋白抗氧化肽的体外抗氧化活性;依次采用离子交换色谱、凝胶色谱对猪皮胶原蛋白酶解液进行分离纯化。结果显示:超滤分离获得510ku的抗氧化肽较其他分子量范围的抗氧化肽具有较好的抗氧化活性;采用离子交换色谱、凝胶色谱两种分离方法分步分离能达到较好的分离纯化效果,离子交换色谱分离得到7个组分,其中组分P1对O-2·清除率最高,多肽浓度为0.85mg/mL时,清除率为46.30%,IC50值为1.24mg/mL;该组分经过凝胶色谱分离后得到2个组分,其中组分P1-B对O-2·清除率最高,多肽浓度为0.9mg/mL时,清除率为49.43%,IC50值为0.98mg/mL。Abstract: Alkaline protease were used to hydrolyze pigskin collagen protein to get antioxidant peptides.And to get the one with high antioxidant activity and purity, a variety of evaluative systems in vitro had been used to research the antioxidant activity of antioxidant peptides with different molecular weights which were obtained through ultrafiltration.At the same time, ion exchange chromatography and gel permeation chromatography were used to separate and purify the enzymolysis solution.It showed that: the antioxidant activity of antioxidant peptides with5~10ku which were obtained through ultra-filtration was better compared to other molecular weights.Ion exchange chromatography and gel permeation chromatography could make a good effect on separation and purification, seven components had been separated by ion exchange chromatography, including the component P1 with the highest scavenging rate on O-2·of 46.30% and IC50 of 1.24 mg /mL when the concentration of polypeptide was0.85 mg /mL.And then, the component P1 was purified into two components by gel chromatography, in which component P1-Bwith the highest scavenging rate on O-2·of 49.43% and IC50 of 0.98 mg /mL when the concentration of polypeptide was 0.9mg /mL.
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