刺参肠内源酶对体壁胶原蛋白的降解作用

摘要

采用明胶酶谱法检测刺参肠内源酶中胶原降解酶的活性,应用流变仪分析刺参体壁粗胶原纤维的流变特性.通过SDS-聚丙烯酰胺凝胶电泳和TCA可溶性寡肽含量测定方法,考察了刺参肠内源酶对体壁酶促溶性胶原蛋白的降解作用.结果表明,当粗胶原纤维质量分数为3%时,溶胀的粗胶原纤维显示出明显的凝胶特性,经刺参肠内源酶处理后,其凝胶性明显下降.在p H 9.0和40℃的条件下,刺参肠内源酶作用0.5 h后,可显著促进PSC中γ、β及α链的降解;在此过程中,体壁酶促溶性胶原蛋白降解产物中TCA可溶性寡肽含量逐渐增加.研究表明,刺参肠中具有能够降解体壁胶原蛋白的内源酶,并在刺参自溶中发挥重要作用.%The activity of endogenous enzyme was determined by gelatin zymography .The rheological behavior of crude collagen fiber from S . japonicus was analyzed by rheometer .The effects of gut endogenous enzymes on degradation of pepsin-soluble collagen (PSC) were analyzed by SDS-PAGE and TCA-soluble oligopeptide content assay .The swelling of crude collagen fibers showed obvious gel characteristic when the crude collagen fiber mass fraction was 3% ,but the gel properties decreased obviously when treated with endogenous enzymes of Stichopus japonicus .The band intensities of γ-,β-andα-chain in PSC significantly decreased by treated with gut endogenous enzymes at pH 9 .0 and 40 ℃ for 0 .5 h .The content of TCA-soluble oligopeptides in the PSC degradation products increased gradually .The result suggested that S . japonicus contain endogenous enzymes that could degrade collagen in the body wall ,and plays an important role in autolysis of S .japonicus .