豆角过氧化物酶的制备及特性研究.rar

摘要:过氧化物酶（Peroxidase,POD）是氧化酶的一类，广泛存在于动物、植物和微生物体内，可应用于酶免疫的测定、药物诊断、生物传感器、有机合成、水污染治理等[1]。

　　　以新鲜豆角为实验原材料，通过匀浆、硫酸铵盐析、离子交换层析以及凝胶过滤层析后得到了电泳纯的豆角过氧化物酶，跑SDS-PAGE凝胶电泳得到一条单一条带，达到了纯化目的。 

　　　本实验在对豆角POD分离纯化的基础上，对所获得的酶还进行了酶学性质的研究，

稳定性实验显示，该酶在30℃-50℃有较好的稳定性，同时,该豆角过氧化物酶的pH适用范围较广泛，在5.0-9.0的环境下有较好的稳定性。豆角过氧化物酶的稳定性较强。通过实验得知，最适温度为40℃，最适pH为6.0，最适条件下的km值为19.05mmol/L。

　　　另一实验表明不同的化学物质对豆角POD活性的影响情况不同。有机溶剂、部分金属离子(如Mn2+、Fe2+)等对该酶有较强的抑制作用，而低浓度草酸、Zn2+和Mg2+对该酶起到一定激活作用。

关键词：过氧化物酶；酶学性质；分离纯化

Abstract:Peroxidase is a sort of oxydase,which can be widely found in animals,plants and microbes,it has been widely used in many fields,such as identify immunity of enzyme,diagnose medicines,biological sensors,organic synthesize,control polluted water,etc.

　Electrophoresis-purity peroxidase from green beans was obtained sequentially after homogenization,extraction,ammonium sulfate precipitation,chromatography and gel filtration chromatography.I achieve the goal to purify peroxidase by SDS-PAGE as a single band.

　A brief protocol for purification of enzyme was worked out in my previous work.Now,a systematic evaluation of the biocatalytic properties of POD was carried out in this study.The experiment of stability shown that the POD was stable under 30℃~50℃, at the same time,the  peroxidase was able to retain its catalytic activity under wide of pH,it was stable under pH 5.0~9.0.Through the experiment,we know that its optimum temperature and pH were 40℃ and 6.0,its Km was determined to be 19.05 mmol/L under optimum conditions.  

　Another experiment which shown that different chemicals have different effect on the activation of peroxidase.Its activity could be strongly inhibited by Organic solvent and part of metals (Mn2+ and Fe2+),but activated by the disulfide bond, Zn2+ and Mg2+ .

Keywords :Peroxidase,POD  Enzymatic properties  Separation and Purification