| 菲律宾蛤仔碱性磷酸酶的分离纯化及部分性质研究 |
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| 引用本文： | 焉翠蔚,随建强,王镇,赵庆娅,黎玲屹.菲律宾蛤仔碱性磷酸酶的分离纯化及部分性质研究[J].中国海洋大学学报(自然科学版),2012(Z2):33-37. |
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| 作者姓名： | 焉翠蔚 随建强 王镇 赵庆娅 黎玲屹 |
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| 作者单位： | 中国海洋大学海洋生命学院 |
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| 基金项目： | 青岛市科技计划基础研究项目(11-2-4-1-(9)-gch);中国海洋大学本科生研究发展计划项目(1112010514)资助 |
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| 摘 要： | 经Tris-HCl缓冲液浸提、硫酸铵分级沉淀盐析、DEAE-Sepharose Fast Flow离子交换层析和Sephadex G-75凝胶过滤层析等步骤,从菲律宾蛤仔内脏团中分离得到碱性磷酸酶,SDS-PAGE显示为单一条带,提纯倍数为14.42,比活力达到131.08U/mg。酶学性质研究表明:该酶亚基分子量约44kD,最适反应温度为45℃,最适pH值为9.0,米氏常数Km为0.098mmol/L,最大反应速度Vmax为1.01μmol/(mL·min)。Ca2+、Mg2+对酶活力表现出显著的激活作用。
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| 关 键 词： | 菲律宾蛤仔 碱性磷酸酶 分离纯化 酶学性质 |
Purification and Partial Characterization of Alkaline Phosphatase from Ruditapes philippinarum |
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| YAN Cui-Wei,SUI Jian-Qiang,WANG Zhen,ZHAO Qing-Ya,LI Ling-Yi.Purification and Partial Characterization of Alkaline Phosphatase from Ruditapes philippinarum[J].Periodical of Ocean University of China,2012(Z2):33-37. |
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| Authors: | YAN Cui-Wei SUI Jian-Qiang WANG Zhen ZHAO Qing-Ya LI Ling-Yi |
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| Institution: | (College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China) |
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| Abstract: | An alkaline phosphatase from the visceral mass of Ruditapes Philippinarum was purified by the following procedures: Tris-HCl buffer extraction,ammonium sulfate precipitation,anion exchange chromatography of DEAE Sepharose Fast Flow,and gel filtration chromatography of Sephadex G-75.The purification factor of the enzyme was 14.42.The specific activity was 131.08U/mg.The purified enzyme showed a single band by SDS-PAGE,with a relative molecular mass of 44 kD.The enzyme exhitited its maximal actitity at pH 9.0 and 45 ℃.The Km and Vmax of the enzyme for PNPP were 0.098 mmol/L and 1.01 μmol/(mL·min) respectively.Ca2+ and Mg2+ had stimulating effects on enzyme activity. |
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| Keywords: | Ruditapes philippinarum alkaline phosphatase purification enzymatic characterization |
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