| 锯缘青蟹碱性磷酸酶分离纯化及部分理化性质研究 |
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| 引用本文： | 陈清西,张喆,庄总来,陈祥仁.锯缘青蟹碱性磷酸酶分离纯化及部分理化性质研究[J].海洋与湖沼,1998,29(4):362-367. |
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| 作者姓名： | 陈清西 张喆 庄总来 陈祥仁 |
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| 作者单位： | 厦门大学生物系!厦门，361005，厦门大学生物系!厦门，361005，厦门大学生物系!厦门，361005，厦门大学生物系!厦门，361005 |
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| 基金项目： | 国家自然科学基金!39470561 |
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| 摘 要： | 于１９９５年２月在厦门海域采集锯缘青蟹，取其内脏经正丁醇抽提，硫酸铵分级分离ＤＥＡＥ－５２离子交换柱层析及ＳｅｐｈａｄｅｘＧ－２００凝胶过滤柱层析纯化，获得碱性磷酸酶 制剂，以快速 蛋白液相色谱及聚丙烯酰胺凝胶电泳检验萁 纯度，获得单一蛋白纯的酶 制剂。
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| 关 键 词： | 锯缘青蟹 碱性磷酸酶 分离 纯化 理化性质 |
| 收稿时间： | 1995/9/10 0:00:00 |
| 修稿时间： | 2/8/1998 12:00:00 AM |
STUDIES ON ISOLATION, PURIFICATION AND SOME PHYSICAL AND CHEIVHCAL PROPERTIES OF ALKALINE PHOSPHATASE
FROM GREENCRAB (SCYLLA SERRATA) |
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| CHEN Qing-xi,ZHANG Zhe,ZHUANG Zong-lai and CHEN Xiang-ren.STUDIES ON ISOLATION, PURIFICATION AND SOME PHYSICAL AND CHEIVHCAL PROPERTIES OF ALKALINE PHOSPHATASE
FROM GREENCRAB (SCYLLA SERRATA)[J].Oceanologia Et Limnologia Sinica,1998,29(4):362-367. |
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| Authors: | CHEN Qing-xi ZHANG Zhe ZHUANG Zong-lai and CHEN Xiang-ren |
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| Affiliation: | Department of Biology Xiamen University, Xiamen, 361005;Department of Biology Xiamen University, Xiamen, 361005;Department of Biology Xiamen University, Xiamen, 361005;Department of Biology Xiamen University, Xiamen, 361005 |
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| Abstract: | An alkaline phosphatase from green crab (Scylla serrata) was collected from Xiamen seawater in February 1995. It was prepared and purified by means of the following techniques: n-butanol extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE cellulose column (DEAE-52) and gel filtration chromatography on Sephadex G-200. The preparation was shown to be homogenous on FPLC and polyacrylarnide gel electrophoresis. The characteristic peak of UV-absorption spectrum of the enzyme was found to be at 278 nm, the fluorescence excitation spectrum at 282 nm, and the fluorescence emission spectrum at 343 nm. The molecular weight of the enzyme is 78 kD. At pH = 9.5, the optimum pH is 9.2, at 37°C. At pH = 9.0, 37°C, the Michaelis constant (KM) is 6.67×10-4 mol/L; Kcat, 460 min-1. Magnesium ion activates the enzyme significantly. Copper ion, mercury ion, methanol, ethanol and ethylene glycol inhibit the enzyme activity in varying degrees, which indicates that the conformation of the enzyme caused by the effectors changes at different levels. The inhibition mechanism had been preliminarily studied. |
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| Keywords: | Scylla serrata Alkaline phosphatase Isolation Purification Physical and chemical properties |
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