对以脱盐乳清蛋白粉为原料的乳杆菌A-2代谢产物中铜离子螯合活性多肽进行分离纯化并进行其抗氧化活性的评价。采用铜离子固定化金属离子亲和层析柱分离,筛选具有铜离子螯合活性多肽,得组分F1、F2、F3,分别经Sephadex G-15凝胶过滤柱层析经进行脱盐分离,得到组分F1-1、F1-2、F2-1、F2-2、F3-1、F3-2,采用ABTS法与抗氧化指数(oxygen radical absorbance capacity,ORAC)法进行抗氧化活性测定,组分F2-1的清除ABTS阳离子自由基能力IC50值为(3.068±0.15) g/L,组分F3-1的清除ABTS阳离子自由基能力IC50值为(5.510±1.56) g/L;组分F2-1的相对ORAC值为(1 246.59±0.72) μmol TE/g,组分F3-1的相对ORAC值为(518.83±1.15) μmol TE/g。组分F2-1、F3-1与铜离子螯合率分别为(59±1.45)%和(6±0.32)%。结果表明,组分F2-1、F3-1具有铜螯合活性组分,具有良好的抗氧化活性。该实验为合理开发乳清蛋白,提供了有效依据。
Isolation and purification of copper chelating polypeptide from Lactobacillus A-2 metabolites using desalted whey protein powder as raw material and evaluation of its antioxidant activity were carried out. Immobilized Cu2+ affinity chromatography column was used to separate Cu2+ chelating peptides, and components F1, F2 and F3 were obtained, and then F1, F2 and F3 were separated by using Sephadex G-15 gel filtration column chromatography, the components F1-1, F1-2, F2-1, F2-2, F3-1 and F3-2 were obtained. The antioxidant activity of the components was determined by the ABTS assay and ORAC assay. The IC50 of F2-1 and F3-1was (3.068±0.15) and (5.510±1.56) g/L, respectively. The relative ORAC value of F2-1 was (1 246.59±0.72) μmol TE/g, that of F3-1 was (518.83±1.15) μmol TE/g. The chelation rates of components F2-1 and F3-1 with Cu2+ were (59±1.45)% and (6±0.32)%, respectively. It was showed that F2-1 and F3-1 had copper chelating activity and good antioxidant activity, which provides an effective basis for the rational development of whey protein.
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