摘要
采用硫酸铵分级盐析、DEAE -Cellulose - 5 2离子交换柱层析、SephadexG - 15 0分子筛柱层析分离纯化了双孢蘑菇 (Agaricusbisporus)子实体超氧化物歧化酶 .纯化了 31倍 ,回收率为 10 .5 1% ,纯酶比活力为 5 5 12 .6u/mg ,酶的最适作用温度为 2 5℃ ,最适 pH值为 8.0 ,酶在 2 5℃以下比较稳定 ,亚基分子质量为 2 1kD ,全酶分子质量为 4 3kD ,该酶由 2个相同的亚基组成 .
The SOD of Agaricus bisporus was purified by salting out with ammonium sulfate ,column chromatography on DEAE-cellulose-52 , gel filtration on Sephadex G-150.The enzyme was purified with only a single band in non denaturing polyacrylamide gel electrophoresis , 31-fold purification,10.51% recovery of enzyme activity and 5 512.6 u/mg specific activity were obtained. The result showed that the enzyme was a kind of Mn-SOD composed of two equally sized subunits ,with a molecular weight of 43 kD, the subunit molecular weight was 21 kD.The optimum temperature and optimum pH for enzyme action were 25 ℃ and pH 8.0 respectively. The enzyme activity was stable under the temperature below 25 ℃,The K m and V max were 0.022 7mol/L and 66.7 mol/min respectively.
出处
《河北大学学报（自然科学版）》
CAS
2002年第3期264-267,共4页
Journal of Hebei University(Natural Science Edition)
基金
全国大学生"挑战杯"基金项目
