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標題: | 水稻酸性磷酸?的純化與定位 Purification and Localization of Acid Phosphatase in Rice |
作者: | 陳武聖 |
出版年 : | 1991 |
學位: | 碩士 |
摘要: | 水稻酸性磷酸?屬多型性,各種同功?等電站皆落在4.5-6.5之間。利用硫酸銨沉澱、膠體過濾層析、集焦管柱層析及聚丙烯醴胺電泳分離純化,並以膠體過瀘層析估計酵素活性分子量在130kd附近。SDS-PAGE分析得有70kd及62kd次體,推測屬異元雙體結構。 部份純化的酸性磷酸?進行特性探討,其活性最適酸鹼度在5.0左右。不同受質的測試以para-nitrophenyl phosphate反應最佳,km值為0.69mM。在60℃下加熱5分鐘,酵素活性降低50%左右。HnSO4促進酵素活性約50%,而HgCl2則抑制90%的活性。用細胞化學定位檢出根冠區中酵素主要聚集在細胞壁及細胞核。外層遊離的細胞則各種胞器都有酸性磷酸?存在。根部分生組織內酵素主要分佈於細胞壁。延長區內以液胞、細胞膜以及細胞核較顯著。至於葉片組織中酸性磷酸?在表皮、葉肉及即將瓦解的細胞均有出現。有關酸性磷酸?的次細胞分佈及其生理意義則在文中進一步討論。 Acid phosphatase (EC 3.1.3.2) extracted from rice seedlings consisted of more than ten kinds of isozymes in zymograni of polyacrylamide gel electrophoresis.They were stepwisely purified by using ammonium sulfate precipitation. Ultro gel filtration, DEAE-trisacryl column and chromatofucsing gel chromatographies. One of the isozymes with optimal pH at 5.0, possessed high affinity to p-nitrophenyl phosphate. FPLC separation showed that its molecular weight was around l30kd, and SDS-PAGE showed that it contained equivalent peptide subunits of apparent Mr 70 and 62 kd. The purified acid phosphatase is heat labile, and its Km is 0.69mM at pH 5.0. Mn•enhances the activity of acid phosphatase, and Hg•is the strongest inhibitor and NaF has only partial inhibitorily effect. Localization of acid phosphatases b the method of Goniori showed that they mainly distributed in cell wall, nucleus and vacuole in root cap. In ground meristem, acid phosphatases were present in cell wall, and in the region of elongation, they were found in tonoplast, cell membrane and nucleus. While in leaf, acid phosphatases appared in vacuole, oil drop, starch grain and cytoplasm. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75783 |
全文授權: | 未授權 |
顯示於系所單位: | 植物科學研究所 |
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