摘要:本实验主要通过匀浆、浸提、离心、硫酸铵分级沉淀、CM-Sepharose离子交换层析、SephaderG-25分子筛凝胶过滤层析等步骤,获得电泳纯的菊芋过氧化物酶(POD),该酶分子量约为37KD。该酶的比活力为12436.92U/mg,纯化倍数为79.45,回收率为2.2%。通过对菊芋过氧化物酶部分理化性质的研究,得到相关数据:该酶的最适反应温度为40℃、最适反应pH值为9.0;该酶在20~60℃以及pH5.0~10.5有较好的稳定性;Cu2+、Ca2+、Zn2+等对该酶有较强激活作用,CTAB、SDS及Mg2+、Mn2+、Fe2+、Al3+等对该酶有较强的抑制作用。结果表明:菊芋过氧化物酶是一种良好的耐酸碱、耐金属离子、常温下活性较高的过氧化物酶。
关键词 菊芋;过氧化物酶;分离纯化;理化性质
Abstract:Electrophoresis-pure peroxidase (POD) from Jerusalem artichoke was obtained through sample homogenization, extraction, centrifugal, ammonium sulfate precipitation, CM-Sepharose chromatography and SephaderG-25 molecular sieve gel filtration. The molecular weight of this enzyme was 37kD. The purified POD had an activity of 12436.92U/mg, the purification factor was 79.45, and the recovery was 2.2%. Some relevant data was obtained by studying the physical and chemical properties of Jerusalem artichoke peroxidase: Its optimum temperature and pH 40℃and 9.0. The POD enzyme was stable under 20~60℃ and pH 5.0~10.5 .Its activity could be strongly activated by Cu2+, Ca2+ and Zn2+, but inhibited by CTAB, SDS, Mg2+, Mn2+, Fe2+ and Al3+. The results showed that: Jerusalem artichoke peroxidase has a good resistance to acid and alkali and metal ions, and higher and activity at room temperature.
Keywords Jerusalem artichoke peroxidase separation of purified physical and chemical properties