摘要： 本论文对马铃薯蛋白的提取及酶解制备多肽进行了初步研究，确定最佳的提取和酶解工艺条件。以新鲜马铃薯为原料，采用盐溶碱提酸沉法提取马铃薯蛋白，在单因素实验的基础上，设计五因素四水平的正交实验，确定马铃薯蛋白提取的最佳工艺参数为：料水比为1:10，NaCl浓度为0.5mol/L，浸提温度60 oC，浸提时间1.5h，pH值为7。该条件下马铃薯蛋白提取率可达0.65%。以提取的马铃薯蛋白为原料，采用酶解法将马铃薯蛋白转化为活性肽。对酶解效果的评价以测定酶解液的DPPH自由基清除率为标准。通过优化实验确定酶解生产活性肽的最佳工艺参数为：酶解时间为90min，底物质量浓度为4g/100mL，加酶量为4%。在此酶解条件下，马铃薯蛋白酶解液的抗氧化活性最高，DPPH自由基清除率可达53.20%。

关键词： 马铃薯蛋白；提取工艺；蛋白酶解；活性肽

Optimization design of extraction and enzymolysis of potato protein to produce active peptides

Abstract: In this paper, the extraction and enzymatic hydrolysis of potato protein were studied preliminarily, and the optimum technological conditions were determined. Using fresh potato as raw material, potato protein was extracted by salt-soluble alkali extraction and acid precipitation method. Based on single factor experiments, orthogonal analysis experiments with five factors and four levels were designed. The optimum technological parameters of potato protein extraction were determined as follows: material-water ratio was 1: 10, NaCl concentration was 0.5 mol/L, extraction temperature was 60 oC, extraction time was 1.5 h, and pH value was 7. Under these conditions, the extraction rate of potato protein could reach 0.65 %. Potato protein was used as raw material to convert potato protein into active peptide by enzymatic method. The evaluation of the enzymatic hydrolysis effect is based on the determination of DPPH radical scavenging rate of the enzymatic hydrolysate. Through optimization experiments, the optimum technological parameters for producing active peptides by enzymolysis were determined as follows: enzymolysis time was 90 min, substrate mass concentration was 4 g / 100 ml, and enzyme dosage was 4 %. Under these conditions, the antioxidant activity of potato protease hydrolysate was the highest, and the DPPH radical scavenging rate could reach 53.20 %.

Keywords: potato protein; extraction process; protease hydrolysis; active peptide

目录S

1 前言 1

1.1 马铃薯 1

1.1.1 马铃薯概述 1

1.1.2 马铃薯蛋白的研究现状 1

1.1.3 马铃薯蛋白的营养价值分析 1

1.1.4 马铃薯蛋白的应用前景 1

1.2 肽 2

1.2.1 肽的概述 2

1.2.2 肽的制备 2

1.2.3 肽的特性及生理功能 2

1.3 本课题的研究目的和意义 2

2 材料与方法 4

2.1 材料 4

2.1.1 原料与主要试剂 4

2.1.2 仪器与设备 4

2.2 实验方法 4

2.2.1 马铃薯蛋白的提取 4

2.2.2 马铃薯蛋白提取的单因素实验 5

2.2.3 马铃薯蛋白提取正交实验设计 5

2.2.4 马铃薯蛋白提取得率的测定 6

2.2.5 马铃薯蛋白质的酶解 6

2.2.6 酶解制备多肽的单因素实验 6

2.2.7 酶解制备多肽正交实验设计 7

2.2.8 DPPH自由基清除率的测定方法 7

3 结果与讨论 8

3.1 马铃薯蛋白提取的单因素实验 8

3.1.1 料水比对马铃薯蛋白提取率的影响 8

3.1.2 NaCl浓度对马铃薯蛋白提取率的影响 9

3.1.3 浸提温度对马铃薯蛋白提取率的影响 9

3.1.4 浸提时间对马铃薯蛋白提取率的影响 10

3.1.5 pH值对马铃薯蛋白提取率的影响 10

3.2 马铃薯蛋白提取条件优化的正交实验 11

3.3 酶解制备多肽的单因素实验 13

3.3.1 底物质量浓度的影响 13

3.3.2 加酶量的影响 14

3.3.3 酶解时间的影响 14