鳗鱼钙螯合肽制备工艺研究

Study on the Preparation for Calcium-chelating Peptides from Eel Protein

摘要:

本文以鳗鱼尾为原料，采用现代酶解技术高效制备鳗鱼钙螯合肽。系统研究了单酶的种类、多酶复配、酶解时间和酶添加量对蛋白回收率、肽得率以及产物钙螯合能力的影响规律，并获得最佳酶解工艺：添加鳗鱼尾蛋白3.5‰的胰酶，调整pH至8.0在55 ℃下酶解3 h，然后调整体系pH至4.0，加入鳗鱼尾蛋白3.5‰的酸性蛋白酶在55 ℃下继续酶解2 h得到鳗鱼肽，蛋白回收率和肽得率分别为66.61%和52.46%。并在此基础上，研究了螯合温度、pH值以及质量比对鳗鱼肽螯合能力的影响，优化获得螯合条件为：鳗鱼肽在30 ℃，pH值为9.0条件下按肽钙质量比1:2，与钙离子螯合30 min得到鳗鱼钙螯合肽，螯合物的钙离子含量为56.78 mg/g。通过氨基酸成分分析，发现鳗鱼肽与钙的螯合反应中，谷氨酸和天冬氨酸等酸性氨基酸的贡献最大。

Abstract:

In this study, the calcium-chelating peptide was prepared from eel tail protein by using modern enzymolysis technology. Effects of different single and multiple proteases, enzymolysis time and addition levels of enzyme on protein recovery, peptides yield, calcium chelating ability were systematically evaluated, respectively. The optimal enzymolysis process was obtained as follows: trypsin hydrolysis (3.5‰) was performed at pH 8.0 and 55 ℃for 3 h, and then the pH of the mixture was adjusted to 4.0, followed by acid protease (3.5‰) hydrolysis for 2 h at 55 ℃to obtain eel peptides. The protein recovery and peptides yield were 66.61% and 52.46%, respectively. The effects of chelating temperature, pH value and mass ratios on the calcium chelating ability of eel peptides were investigated. The chelating conditions of calcium-chelating peptides were optimized as follows: 30 ℃, pH 9.0, the mass ratio of eel peptides to calcium 1:2 and chelating with calcium for 30 mins. The calcium content of calcium-chelating peptides was 56.78 mg/g. In addition, glutamic acid and aspartic acid were found to be a great contribution to chelating reaction between eel peptides and calcium via the amino acid composition analysis.