胰蛋白酶对酸奶水解度及酪蛋白磷酸肽生成量的影响—毕业论文.docVIP

摘 要 本实验以脱脂乳粉为原料，首先采用德氏乳杆菌保加利亚亚种(L18)和嗜热链球菌(St)制备酸奶，以水解度为指标探讨胰蛋白酶水解酸奶的最佳酶解条件及在该条件下CPPs的生成量，其中CPPs的制备和含量的测定分别采用胰蛋白酶水解法和分光光度法。 胰蛋白酶水解纯奶和酸奶的最佳酶解条件为：蛋白溶液浓度3%、pH8.0、在37℃，酶:底物=1.00:100，时间为3h，在该条件下纯奶的水解度为20.40%，制备CPPs的量为每克蛋白质146mg，酸奶的水解度为27.55%，制备CPPs的量为每克蛋白质207mg。此时N/有机P比值为8.20。 关键字：酪蛋白磷酸肽；水解度；德氏乳杆菌保加利亚亚种；嗜热链球菌 Abstract In this study，firstly, skim milk powder was used to prepare yogurt by mixing fermentation of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus.The degree hydrolysis experiments were found to discuss the optimum conditions of hydrolysis for pure milk and yogurt by trypsin as well as it had effect on the production of CPPs under optimum conditions. CPPs were prepared by the methods of calcium-ethanol precipitation and the contents were determined by spectrophotometry. The hydrolysis experiments demonstrated the best conditions for hydrolysis of pure milk and yogurt by trypsin. The conditions inculded the ratio of enzyme to substrate which was 1:100, the reaction time which was 3 hours, and the hydrolysis reaction which occurred at 37℃ and pH 8.0. Under this conditions the degree of hydrolysis respectively in pure milk and yogurt were 20.40%, 27.55%, and the yield of CPPs were 146mg/g, 207mg/g. Key Words: Casein phosphopeptides;Lactobacillus delbrueckii subsp Bulgaricus;Streptococcus thermophilus;The degree of hydrolysis 目 录 1 引言 1 1.1 酸奶 1 1.1.1 酸奶的定义 1 1.1.2 酸奶的营养价值 1 1.1.3 酸奶的保健功能 2 1.2 乳源生物活性肽 2 1.3 酸奶中的CPPs 3 1.4 研究意义和主要内容 3 2 材料与方法 4 2.1试验材料、主要试剂和仪器······························4 2.1.1 试验原料 4 2.1.2 试验菌种 4 2.1.3 培养基 4 2.1.4 主要化学试剂 4 2.1.5 试验仪器 4 2.2 试验方法 4 2.2.1 菌种活化 4 2.2.2 发酵剂的制备 4 2.2.3 酸奶的发酵工艺 5 2.2.4 胰蛋白酶水解样品制备CPPs 5 2.3 磷（P）含量的测定 5 2.3.1 无机磷含量的测定 5 2.3.2 总磷含量的测定 5 2.3.3 P的标准曲线的绘制 5 2.3.4 有机磷的计算 6 2.3.5 r（N／P）的计算 6 3 结果与分析 6 3.1 时间对酸奶和纯牛奶水解度的影响 6 3.2 酶:底物对水解度的影响 7 3.2.1 酶:底物对纯奶水解度的影响 7 3.2.2 酶:底物对酸奶水解度的影响 8 3.3 P含量的标准曲线 9 3.4 酸奶和纯牛奶来源的CPP