复合酶解可溶性蛋膜蛋白制备多肽工艺优化_周艳华.pdfVIP

92 2010, Vol. 31, No. 02 食品科学 ※工艺技术 复合酶解可溶性蛋膜蛋白制备多肽的工艺优化 周艳华，马美湖* ，蔡朝霞，李 涛，李凤玲 (华中农业大学 国家蛋品加工技术研究分中心，湖北武汉 430070) 摘 要：为研究蛋膜蛋白的利用，采用碱性蛋白酶、木瓜蛋白酶、胰蛋白酶、中性蛋白酶及胃蛋白酶进行单一 酶解筛选实验，然后进行复合酶解实验。采用二次正交旋转组合设计，以水解度、氮收率为指标，研究酶制剂 种类、加酶方式、复合酶比例、总加酶量、酶解时间、p H 值及温度对制备多肽工艺的影响。综合考虑水解度 和氮收率因素，最终确定复合酶解可溶性蛋膜蛋白制备多肽的最佳工艺条件为：总加酶量16000U/g ，并以碱性蛋 白酶与木瓜蛋白酶的酶活力配比为8 :2 先后加入；酶解时间为碱性蛋白酶2h 、木瓜蛋白酶1h ；p H 值为碱性蛋白酶 9.0 、木瓜蛋白酶5.5 ；酶解温度为50 ℃。该条件下制备的蛋膜蛋白酶解产物水解度和氮收率分别为46.12%、85.56% 。 关键词：可溶性蛋膜蛋白；复合酶解；多肽；二次回归正交旋转组合设计 Quadratic Orthogonal Rotation Combination Design-based Optimization of Enzymatic Production of Polypeptides from Soluble Eggshell Membrane Protein ZHOU Yan-hu a ，MA Mei-hu * ，CAI Zhao-xia ，LI Tao ，LI Fen g-ling (National Research and Development Center for Egg Processing, Huazhong Agricultural University, Wuhan 430070, China) Abstract ：In this study, eggshell membrane was used as the raw material to prepare polypeptides by enzymatic hydrolysis. In order to maximize degree of hydrolysis (DH) and nitrogen yield, the optimal hydrolysis mode of eggshell membrane was selected from hydrolysis with pairwise combination (used sequentially or simultaneously) of alkaline protease, papaya protease and trypsin that resulted in higher degree of hydroly sis and nitrogen yield than neutral protease and pep sin , or each of them. Meanwhile, based on this, the optimal values of crucial technological parameters, including temperature, hydrolysis time, pH and total enzyme dose, were determined by the method of quadratic orthogonal rotation combination design. Results showed that the optimal enzymatic production process was based on sequential hydrolysis at 50 ℃with alkaline protease for 2 h at pH 9.0 followed by pa