水解面筋蛋白高效制备谷氨酰胺结合肽的研究-食品科学.PDF

※工艺技术 食品科学 2006, Vol. 27, No. 09 181 水解面筋蛋白高效制备谷氨酰胺结合肽的研究 杨得胜，张源淑，邹思湘* ，陈伟华 (南京农业大学 农业部动物生理生化重点开放实验室，江苏 南京 210095) 摘 要：采用食品级胃蛋白酶、胰蛋白酶、Protamex 复合酶双酶组合对面筋蛋白进行水解制备Gln 结合肽。先确 定三种单酶各自对面筋蛋白适宜的初始pH 、温度、酶浓度和底物浓度。并以双酶进行两两组合水解。在进行水 解度控制实验时取样，分析水解液中氨基氮的含量，计算平均肽链长度，并用高效液相色谱分析水解液中Gln 的 得率。实验结果表明胰蛋白酶和胃蛋白酶联合水解的效果最好，条件为：胰蛋白酶在pH8.0 ，50℃，S%=12.8% (W/V)，E%=9%(W/W)水解6h ，再用胃蛋白酶pH2.0 ，40 ℃，E%=5%(W/W)条件下水解5h ，可得到平均肽链长 度为2.20 个氨基酸残基的蛋白水解液，Gln 得率最高，为60.43%。认为胰蛋白酶和胃蛋白酶组合是面筋蛋白中高 效制备谷氨酰胺结合肽的最佳组合。 关键词：谷氨酰胺；谷氨酰胺结合肽；蛋白酶组合 EffectiveProcessingof Gln-boundPeptidefromWheatGluten YANG De-sheng，ZHANG Yuan-shu，ZOU Si-xiang*，CHEN Wei-hua (KeyLabofAnimalPhysiologyandBiochemistry,Ministry ofAgriculture,NanjingAgriculturalUniversity, Nanjing 210095, China) TM Abstract：The food grade proteases such as Pepsin, Trypsin, Protamex were twined to prepare high yield of Gln-bound TM peptidesfromwheat gluten.AtfirstTrypsin,PepsinandProtamex wererespectivelydeterminedtheirproper initialpHvalue, temperature, enzyme concentration and substrate concentration for wheat gluten hydrolysis. Then the wheat gluten was hydrolyzed with enzyme twined. The amino nitrogen content and the average amino acid numbers of peptides in hydrolysates were analysed, and the yield of Gln was determined by HPLC. The result showed that the combination of Trypsin and Pepsin performed a better hydrolytic effect on gluten.The optimum conditions are as follows: Trypsin atpH8.0, 50℃, S%=12.8% (W/V) and E%=9%(W/W) for 6 hours, then Pepsin at pH2.0, 40℃ and E%=5%(W/W) for 5 hours. The final average amino acid