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    豆角過氧化物酶的制備及特性研究.rar

    資料分類:農業大學 上傳會員:coffee45 更新時間:2014-11-03
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    摘要:過氧化物酶(Peroxidase,POD)是氧化酶的一類,廣泛存在于動物、植物和微生物體內,可應用于酶免疫的測定、藥物診斷、生物傳感器、有機合成、水污染治理等[1]。

       以新鮮豆角為實驗原材料,通過勻漿、硫酸銨鹽析、離子交換層析以及凝膠過濾層析后得到了電泳純的豆角過氧化物酶,跑SDS-PAGE凝膠電泳得到一條單一條帶,達到了純化目的。 

       本實驗在對豆角POD分離純化的基礎上,對所獲得的酶還進行了酶學性質的研究,

    穩定性實驗顯示,該酶在30℃-50℃有較好的穩定性,同時,該豆角過氧化物酶的pH適用范圍較廣泛,在5.0-9.0的環境下有較好的穩定性。豆角過氧化物酶的穩定性較強。通過實驗得知,最適溫度為40℃,最適pH為6.0,最適條件下的km值為19.05mmol/L。

       另一實驗表明不同的化學物質對豆角POD活性的影響情況不同。有機溶劑、部分金屬離子(如Mn2+、Fe2+)等對該酶有較強的抑制作用,而低濃度草酸、Zn2+和Mg2+對該酶起到一定激活作用。

    關鍵詞:過氧化物酶;酶學性質;分離純化

     

    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

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    上傳會員 coffee45 對本文的描述:過氧化物酶是一種血紅素蛋白,由單一肽鏈和卟啉組成,血紅素蛋白與脫輔基蛋白分子組成全酶分子。過氧化物酶蛋白的分子質量約為35kD,含有約氨基酸殘基序列300個左右,有血紅素和......
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