文档介绍：大蒜SOD酶性质研究
摘要:超氧化物歧化酶(Superoxide Dismutase,SOD)是生物体内专一清除超氧阴离子的蛋白酶。自然界中SOD分布极广,其含量随生物体的不同而不同,即使同一种生物的不同组织或同一组织的不同部位,其SOD的种类和含量也有很大差别。
大蒜是SOD含量较丰富的天然植物之一。本论文探讨了从大蒜中提取与纯化SOD的工艺路线、优化条件、纯化效果及大蒜SOD的生化性质。实验结果表明,经过三次纯化,大蒜SOD的比酶活有显著提高;且大蒜SOD具有较好的热稳定性,在60℃时酶活达到最大,总体呈先增后减的趋势。
关键词:大蒜;超氧化物歧化酶;纯化;性质
Abstract:Superoxide Dismutase is a kind of important enzyme anism,which can specifically eliminate the superoxide anion. SOD are very widespread in nature, and its content varies with anism, even with a biological or anizations in different parts of the anization, the type and content of SOD is also very different.
Garlic is rich in SOD content of natural plants. This thesis discusses the process route ,optimum conditions
,result of purification about the extraction and purification of SOD from garlic and the biochemical properties of garlic SOD. Experimental results show that after three purification, activity of garlic SOD was significantly SOD has good thermal stability and maximum activity at 60 ℃. First increase after reduction of the overall trend showed.
Key words: garlic; superoxide dismutase;purification
; properties
大蒜是人们喜爱的一种香辛蔬菜和调味料,历来被认为具有保健和治疗作用,其活性物质主要是蒜氨酸和超氧化物歧化酶(SOD)。SOD是生物体内产生的超氧阴离子自由基的清除剂,具有抗衰老,提高机体对多种疾病的抵抗力,增强机体对外界环境的适应力,减轻肿瘤患者在放疗、化疗过程中的严重毒副作用等生理功能。所以在医药、食品、化妆品、
农业等方面都具有广泛的应用。大蒜中SOD含量较高,其活性是一般果蔬的数倍,被认为是人类重要的SOD来源。本文以市场上销售的大蒜为材料,通过分离提取大蒜的SOD粗酶,并用氯仿一乙醇沉淀、丙酮沉淀、硫酸铵分级沉淀对SOD粗提物进行分离纯化,测定大蒜不同温度下SOD活力变化,为今后实际研究或提取大蒜的SOD粗酶提供一个选择依据。
市售新鲜大蒜
(1)( ml/L mL NaH2PO4配置而成);
(2)氯仿一乙醇混合液(氯仿:乙醇为3:5)
(3)标准蛋白质溶液:(用移液枪吸取1ml浓度为1mg/ml标准蛋白质溶液,溶于蒸馏水并定容至10ml,)、
(4)考马斯亮蓝G-250(%)染液:(-250,溶于5ml 90%乙醇中,加入85%(m/v)的磷酸10ml,最后用蒸馏水定容至100ml(此溶液不宜久存,在常温中可放置1~2
个月),将定容好的染液过滤并装入棕色瓶中保存)
(5)130mmol/l甲硫氨酸溶液()
(6)750umol/l NBT( PBS定量至100ML(避光)
(7)100umol/L EDTANA2( EDTANA2,用PBS定量至100ml)
(8)20umol/l核黄素(