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將酸性蛋白酶酶解產(chǎn)物經(jīng)過SP-Sephadex C-25強陽離子交換色譜、Sephadex G-50凝膠過濾色譜和反相高效液相色譜等步驟，從鯊魚皮明膠中分離到一種分子量為1356.3Da的抗氧化肽Ke-ⅱ，當(dāng)其濃度為125μg/ml時，對DPPH自由基的清除率能夠達(dá)到62.20%。 為了進(jìn)一步分離更多的膠原多肽，又采用超濾技術(shù)對酸性蛋白酶酶解產(chǎn)物中分子量小于3500Da的的組分進(jìn)行初分離并濃縮，凍干后繼續(xù)采用和之前相同的色譜手段進(jìn)行分離，最后得到三個抗氧化活性最強的膠原多肽B-Ⅲa 、B-Ⅲc和B-Ⅲd。經(jīng)C18反相高效液相色譜鑒定，這三個多肽組分都基本表現(xiàn)為單一峰。接著，再通過ESI Q-TOF MS對它們進(jìn)行鑒定，結(jié)果表明B-Ⅲa 、B-Ⅲc、B-Ⅲd分別為分子量為944.5、784.5以及812.5的多肽。其中，B-Ⅲd經(jīng)氨基酸N端測序，得知其為含10個氨基酸，序列為Gly-Thr-X-Gly-Ala-Y-Gly-Pro-Z-Gly（未完全公開）的多肽。 |
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ljason
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【答案】應(yīng)助回帖
xukun1176(金幣+100, 翻譯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樓2011-04-13 14:43:36
ljason
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【答案】應(yīng)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樓2011-04-13 15:03:42
qingkong0918
金蟲 (小有名氣)
晴空
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【答案】應(yīng)助回帖
xukun1176(金幣+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樓2011-04-13 15:37:38
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