谷朊粉钙离子螯合肽制备工艺优化及其结构表征

Optimization of preparation process and structural characterization of calcium ion chelated peptide from gluten

中文关键词:  谷朊粉  螯合肽  酶解  结构表征；响应面

英文关键词:gluten  chelated peptide  enzymatic hydrolysis  structural characterization  response surface method

中文摘要:

      目的 优化谷朊粉钙离子螯合肽制备工艺并对其结构进行表征。方法 以水解度为指标, 探究温度、时间、pH及加酶量等因素对谷朊粉酶解的影响; 以钙离子螯合肽生成量为指标, 探究不同因素对钙离子螯合肽生产量的影响; 并根据结果进行Box-Behnken响应面实验优化螯合工艺。通过扫描电镜分析和红外分析对谷朊粉钙离子螯合肽进行表征。结果 最佳酶解工艺为: 液料比17.5:1 (mL/g)、碱性蛋白酶和风味蛋白酶添加比例为3:1、加酶量5000 U/g、酶解pH 9、酶解温度50℃、酶解时间4.0 h; 优化后的最佳螯合工艺为: 螯合温度38℃、肽液浓度13.5%、肽钙比5:1、螯合时间12.0 min、螯合pH 7、静置时间35 min、无水乙醇添加量为7倍体积, 在此条件下钙离子螯合肽得率为50.85%。扫描电镜结果显示, 螯合肽外观发生明显改变, 更有利于吸收; 红外光谱分析表明, 钙离子可能与谷朊粉多肽上的-NH、-COOH结合。结论 本研究建立的谷朊粉酶解工艺、钙离子螯合工艺可行, 可为谷朊粉深加工和产品研发提供思路与理论参考。

英文摘要:

      Objective To optimize the preparation process of calcium ion chelated peptide from gluten and characterize its structure. Methods Using hydrolysis degree as an indicator, the effects of temperature, time, pH, and enzyme dosage on the enzymatic hydrolysis of wheat gluten powder were, explored; the effects of different factors on the production of calcium ion chelated peptide were explored by using the production of calcium ion chelated peptides as an indicator; and based on the results, Box Behnken response surface experiment was conducted to optimize the chelation process. The calcium ion chelating peptide from gluten powder was characterized by scanning electron microscopy and infrared analysis. Results The optimal enzymatic hydrolysis process was as follows: Liquid to material ratio 17.5:1 (mL/g), addition ratio of alkaline protease and flavor protease 3:1, enzyme dosage 5000 U/g, enzymolysis pH 9, enzymolysis temperature 50℃, enzymolysis time 4.0 hours; the optimized chelation process was as follows: Chelated temperature 38℃, peptide solution concentration 13.5%, peptide calcium ratio 5:1, chelated time 12.0 minutes, chelated pH 7, standing time 35 minutes, anhydrous ethanol addition amount 7 times the volume. Under these conditions, the yield of calcium ion chelated peptide was 50.85%. The scanning electron microscopy results showed that the appearance of chelated peptides had undergone significant changes, which was more conducive to absorption; infrared spectroscopy analysis indicated that calcium ions might bind to -NH and -COOH on gluten peptides. Conclusion The enzymatic hydrolysis process and calcium ion chelation process established in this study are feasible, and can provide ideas and theoretical references for the deep processing and product development of gluten.