摘要：马勃为担子菌门真菌类中草药，民间常用于止血、消肿，马勃多糖细胞毒性小。本课题以大秃马勃为研究对象，采用单因素-响应面实验，优化了马勃的提取工艺，在水提法的基础上，比较了纤维素酶、果胶酶对马勃多糖提取的效果，分析提取液的活性成分和生物活性。实验结果表明：水提法提取的最佳工艺条件为料液比1:50、90℃、提取2h，多糖得率为10.21%。酶法辅助能有效提高马勃多糖的收率，采用纤维素酶与果胶酶按3:1复配、加酶量6%，温度90℃、时间2h，多糖的得率为12.73% 较单纯水提法提高了2.52%。马勃提取液含有多肽、多酚、黄酮、蛋白质等多种活性成分，其水提液和马勃多糖溶液具有还原力、DPPH自由基清除能力、羟基自由基清除能力，能有效抑制金黄色葡萄球菌、大肠杆菌、铜绿假单胞菌、酵母菌。

关键词： 马勃；多糖；提取；酶解；抗氧化；抗菌

Optimization of Process Conditions for Extraction of Polysaccharides from Calvatia gigantea by Enzymatic Hydrolysis

Abstract: Calvatia gigantea is a fungal strain of Chinese herbal medicine, commonly used in the folk for hemostasis, detumescence, and the polysaccharide has little cytotoxicity. This topic focuses on the Calvatia gigantea，Single factor-response surface experiment was used to optimize the extraction process of Puffball. Based on the water extraction method, the effect of cellulase and pectinase on the extraction of P. morifolium was compared. Analyze the active ingredients and biological activity of the extract . The experimental results show that the optimum conditions for water extraction are the ratio of material to liquid: 1:50, 90°C, 2h, and the yield is 10.21%. Enzyme-assisted energy can effectively increase the yield of polysaccharides. Cellulase and pectinase are compounded in 3:1 and the amount of enzyme is 6%. The temperature is 90°C and the time is 2h. The yield of polysaccharide is 12.73%.Compared with water extraction, the yield increased by 2.52%.The test results showed that Calvatia gigantea extract contains many active ingredients such as polypeptides, polyphenols, flavonoids, and proteins. The water extract and polysaccharide solution have the reducing power, DPPH free radical scavenging ability, and hydroxyl radical scavenging ability. Effectively inhibit Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, yeast.

Keywords: Calvatia gigantea、polysaccharide、extraction、Enzymatic、antioxidant,、antibacterial

目录

1前言 1

1.1 食用菌多糖的研究与发展现状 1

1.1.1 食用菌多糖的种类 1

1.1.2 食用菌多糖的提取方法 1

1.1.2 食用菌多糖的营养价值和生理活性 2

1.1.3 食用菌多糖的开发价值 3

1.2 马勃多糖的研究进展 3

1.3 课题研究的主要内容与意义 4

2实验材料与方法 5

2.1 实验材料与主要试剂 5

2.2 实验仪器 5

2.3 实验方法与步骤 6

2.3.1 马勃预处理 6

2.3.2 水提工艺条件的优化 6

2.3.3 酶法辅助提取条件的优化 6

2.4 活性分析 6

2.4.1 抗氧化实验 6

2.4.2 抗菌实验 7

2.5 成分分析 7

2.5.1 多糖的测定方法 7

2.5.2 多酚的测定方法 9

2.5.3 黄酮的测定方法 10

2.5.5 多肽含量的测定方法 11

2.5.6 总蛋白质的测定方法 12

3实验结果与讨论 13

3.1 马勃预处理结果 13

3.2 水提法马勃多糖提取工艺条件优化 13

3.2.1 料液比对马勃多糖得率的影响 13

3.2.2 提取温度对马勃多糖提取率的影响 13

3.2.3 提取时间对马勃多糖得率的影响 14

3.2.4响应面实验分析 15

3.3 酶法辅助马勃多糖提取工艺条件优化 18

3.3.1 果胶酶浓度的确定 18

3.3.2 纤维素酶浓度的确定 19

3.3.3 复合酶配比的确定 19

3.3.4 复合酶添加量的确定 20