酪蛋白-胰酶体系多肽释放的色谱表征.docVIP

  酪蛋白-胰酶体系多肽释放的色谱表征# 苏荣欣，殷文文，齐崴，何志敏* （天津大学化工学院化学工程研究所） 5 10 摘要：酪蛋白是抗菌肽、磷酸肽、抗高血压肽等多种活性肽的有效来源。本文研究了 pH8.0 时胰酶在间歇反应器中水解酪蛋白的反应行为，利用反相色谱和离子交换色谱分析了酪蛋白 -胰酶体系的产物多肽，探讨了产物多肽疏水性和荷电性的变化规律。结果表明：各种酪蛋 白单体在反应 30min 后全部水解；在胰酶作用下，疏水性较强的多肽逐渐水解为亲水性多肽； 多电荷多肽逐渐转化为寡电荷多肽，含多个正电荷的抗菌肽含量先增加再减少，而带负电荷 的酪蛋白磷酸肽则随反应的进行逐渐增加。反相色谱和离子交换的色谱行为分析，可指导抗 菌肽、酪蛋白磷酸肽、疏水性多肽和亲水性多肽的酶法制备。 关键词：化学工程；酪蛋白；胰酶；酶促水解；活性多肽；色谱 中图分类号：Q814 15 Chromatographic analysis of released peptides from pancreatic hydrolysis of casein Su Rongxin, Yin Wenwen, Qi Wei, He Zhimin (Chemical Engineering Research Cengter, School of Chemical Engineering and Technology, 20 25 30 35 40 Tianjin University, Tianjin 300072) Abstract: Casein is an effective source of important bioactive peptides with antimicrobial, antihypertensive, metal-binding, and other functions. In this paper, pancreatic hydrolysis of bovine casein was carried out at pH 8.0 in a batch stirred tank reactor. The released peptides were analyzed by reversed phase liquid chromatography (RPLC) and ion exchange chromatography (IEC). The changes in the hydrophobicity and charge states of peptides were also discussed. All the invidual caseins were digested after 30 min hydrolysis. the contents of hydrophobic peptides decreased due to the further hydrolysis to the hydrophilic peptides. The content of antimicrobial peptides containing multiple positive charges increased at first and then decreased, whereas the content of phosphopeptides increased with increasing reaction time. The chromatographic analysis of RPLC and IEC was a theoretical guide for the preparation of antimicrobial, phospho-, hydrophobic and hydrophilic peptides. Key words: Chemical engineering; casein; pancreatin; enzymatic hydrolysis; bioactive peptides; chromatography 0 引言 酪蛋白是由?S1-酪蛋白、?S2-酪蛋白、?-酪蛋白、?-酪蛋白等四种单体聚集而成的胶束 分子。它不仅是一种极具营养价值的蛋白质（含有人体全部必需氨基酸），而且还是近十类 百余种活性多肽的有效肽源[1]（功能涵盖免疫调节、自由基清除、矿物质载体、抗癌、抑菌、 溶栓、降高血压等）。 液相色谱技术是生物分离的主流技术。目前，制备蛋白质、多肽类活性物质主要采用离 子交换色谱、体积排阻色谱和反相色谱。其中，反相色谱是生物复杂样品分离中最常用的分 离手段，具有很强的分离效果和分辨率。而离子交换