赵学兰， 赵　越， 宋成军

(1. 天津生物工程职业技术学院，天津 300074； 2. 环境保护部环境规划院，北京100012； 3. 农业部

规划设计研究院 农村能源与环保研究所 农业部农业废弃物能源化利用重点实验室，北京 100125)



蚯蚓中超氧化物歧化酶(SOD)提取条件的优化研究

赵学兰1， 赵越2， 宋成军3

(1. 天津生物工程职业技术学院，天津 300074； 2. 环境保护部环境规划院，北京100012； 3. 农业部

规划设计研究院 农村能源与环保研究所 农业部农业废弃物能源化利用重点实验室，北京 100125)

摘要超氧化物歧化酶(Superoxide dismutase, SOD)是一种重要的抗氧化剂，在农业和医药领域具有巨大的应用价值。利用磷酸盐缓冲液提取赤子爱胜蚓(Eisenia foetida)中SOD，对H3PO4缓冲液pH值、热处理温度、料液比与(NH4)2SO4饱和度等关键参数设计单因素实验，采用4因素4水平的正交试验法[L(16)(45)]进行优化。结果表明：热处理温度对蚯蚓SOD活性有显著性影响，主次顺序为：热处理温度>料液比>H3PO4缓冲液pH值>(NH4)2SO4饱和度。蚯蚓SOD的最佳提取工艺为： H3PO4缓冲液pH值为8.0、热处理温度为30℃，料液比为6，以及硫酸铵饱和度为51%～70%。正交实验设计优化后，蚯蚓SOD活性达到了2895.32 U/g[FW]。为大规模生产蚯蚓SOD提供了数据支撑。

关键词蚯蚓;超氧化物歧化酶(SOD)；活性；提取条件；正交试验

Optimization of SOD extraction conditions from earthworm

ZHAO Xue-lan1, ZHAO Yue2, SONG Cheng-jun3

(1. Tianjin Vocaltional College of Bioengineering, Tianjin 300074; 2. Chinese Academy for Environmental Planning,Beijing 100012; 3. Key Laboratory of Energy Resource Utilization from Agricultural Residues (MOA), Institute of Energy and Environmental Protection, Chinese Academy of Agriculture Engineering, Beijing 100125, China)

AbstractSuperoxide dismutase (SOD) has been demonstrated to be an important antioxidant, which protects the cells exposed to the toxicity of oxygen and has displayed huge application value and development prospect. The phosphate buffer was used to extract SOD, the antioxidant component of earthwarm (Eiseniafoetida). Four key extraction process parameters selected for single-factor tests were phosphate buffer pH value， heat-treatment temperature， ratio of solid to liquid, and degree of saturation((NH4)2SO4). Four factors and four levels of orthogonal experiment were used to determine the optimum factors levels above. Then activity of the SOD was measured by adjacent benzene three phenolic autoxidation method. Results indicated that the optimal levels of these parameters were as follows: phosphate buffer pH value, 8.0; the heat treatment temperature, 30℃; ratio of solid to liquid 6, and degree of saturation((NH4)2SO4), 51%-70%. Under these conditions, the activity of the SOD was up to 2895.32 U/g[FW]. This study would provide a technical basis for the production of large scale production of SOD from earthworm.

Keywordsearthworm; superoxide dismutase; activity; extraction conditions; orthogonal experiment

世界上有蚯蚓4 000余种[5]，我国已经记录的有9科28属306种[6]。蚯蚓特殊的生活环境赋予其体内超氧化物歧化酶成分含量很高，回收率为35%[7]，蚯蚓SOD活性较其他物种高，每克鲜重高达4 000单位[8]。蚯蚓作为一种中药材入药已有很久的历史，易饲养，价格低，对于大规模生产SOD，是一种很好的原料[9]。

本研究利用磷酸盐缓冲液法提取蚯蚓SOD，再用邻苯三酚自氧化法测定活性[10]。在单因……