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将酸性蛋白酶酶解产物经过SP-Sephadex C-25强阳离子交换色谱、Sephadex G-50凝胶过滤色谱和反相高效液相色谱等步骤，从鲨鱼皮明胶中分离到一种分子量为1356.3Da的抗氧化肽Ke-ⅱ，当其浓度为125μg/ml时，对DPPH自由基的清除率能够达到62.20%。 为了进一步分离更多的胶原多肽，又采用超滤技术对酸性蛋白酶酶解产物中分子量小于3500Da的的组分进行初分离并浓缩，冻干后继续采用和之前相同的色谱手段进行分离，最后得到三个抗氧化活性最强的胶原多肽B-Ⅲa 、B-Ⅲc和B-Ⅲd。经C18反相高效液相色谱鉴定，这三个多肽组分都基本表现为单一峰。接着，再通过ESI Q-TOF MS对它们进行鉴定，结果表明B-Ⅲa 、B-Ⅲc、B-Ⅲd分别为分子量为944.5、784.5以及812.5的多肽。其中，B-Ⅲd经氨基酸N端测序，得知其为含10个氨基酸，序列为Gly-Thr-X-Gly-Ala-Y-Gly-Pro-Z-Gly（未完全公开）的多肽。 |
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xukun1176(金币+100, 翻译EPI+1): 2011-04-19 10:22:28
| 第一段：Antioxidant Peptide Ke-ⅱ with a molecular weight of 1356.3Da was isolated from shark skin gelatin after the characterization SP-Sephadex C-25 strong cation exchange chromatography, Sephadex G-50 gel filtration chromatography and RP-HPLC of acid protease enzyme product. The scavenging ratio could reach 62.20% when its concentration was 125μg/ml. |
2楼2011-04-13 14:43:36
【答案】应助回帖
| In order to further separation more collagen peptides, acid hydrolysis products of molecular weight less than 3500Da was separated and concentrated by ultrafiltration technology. the three strongest antioxidant activity of the collagen peptide B-Ⅲ a, B-Ⅲ c and B-Ⅲ d was obtained via lyophilize and chromatographic separation. the three polypeptide components were performed a single peak when identification by RP-HPLC C18. the molecular weight of the peptide B-Ⅲ a, B-Ⅲ c and B-Ⅲ d were 944.5,784.5 and 812.5 obtained from ESI Q-TOF MS identification. The B-Ⅲ d was contained 10 amino acids detected by amino acid N terminal sequencing and the sequence was Gly-Thr-X-Gly-Ala-Y-Gly-Pro-Z-Gly (not fully open). |
3楼2011-04-13 15:03:42
qingkong0918
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【答案】应助回帖
xukun1176(金币+100): 2011-04-19 10:22:37
| The zymohydrolysised product of acid protease is processed by Strong cation exchange SP-Sephadex C-25 , gel filtration chromatography Sephadex G-50 and reversed-phase high-performance liquid chromatography，and anti-oxidant peptide is obtained，whose molecular weight is 1356.3Da。 Rate of clearing free radical DPPH can reach 62.20% when the concentration is 125μg/ml . In order to separate more collagenous polypeptide， those whose molecular weight are less than 3500Da are separated and condensed initially by hythseperfiltration, then are ayophilized，seperated by the same chromatographs as above, three collagenous polypeptides with the most strong anti-oxidant activity are obtained, B-Ⅲa 、B-Ⅲc和B-Ⅲd. Assessed by C18 reversed-phase high-performance liquid chromatography , the three polypeptides all present simple spike. The molecular weight of the three polypeptides are 944.5、784.5 and 812.5 respectively. Especially , B-Ⅲd is assessed to have 10 amino acids , N terminal amino acid sequence is Gly-Thr-X-Gly-Ala-Y-Gly-Pro-Z-Gly（not fully publicized）. |
4楼2011-04-13 15:37:38