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。
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 Vma. 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.
出处
《中国海洋大学学报(自然科学版)》
CAS
CSCD
北大核心
2012年第7期33-37,共5页
Periodical of Ocean University of China
基金
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
关键词
菲律宾蛤仔
碱性磷酸酶
分离纯化
酶学性质
Ruditapes philippinarum
alkaline phosphatase
purification
enzymatic characterization