摘要
新鲜大豆芽经匀浆、磷酸缓冲液抽提、硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、Superdex-200凝胶过滤层析,获得电泳纯的过氧化氢酶。该酶酶活回收率为30.92%,纯化倍数为293.09,酶比活力达到26 322.43 U/mg。该酶全分子质量为234.9 k D,亚基分子质量为57.42 k D;最适p H值为7.2,最适温度为30℃,在p H 4.0~10.6及25~35℃范围内有较好的稳定性;在30℃、p H 7.2条件下测得该酶对过氧化氢的Km值为31.82 mmol/L;十二烷基硫酸钠、草酸、Ag+、Cu2+对该酶有强烈的抑制作用,Pb2+对该酶有双重作用,低浓度时有激活作用而高浓度时抑制酶活性。
Electrophoresis-purity catalase(CAT) was obtained from freshly sprouted soybean(Glycine max) by the sequential steps of homogenization, buffer solution extraction, ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Superdex-200 gel filtration chromatography. The purified enzyme had a specific activity of 26 322.43 U/mg, with a 30.92% activity recovery and a 293.09-fold purification. The molecular mass of the CAT enzyme comprising a 57.42 k D subunit was 234.9 k D. Its optimum p H and temperature were 7.2 and 30 ℃, respectively. This enzyme was stable at p H 4.0–10.6 and at 25–35 ℃. Its Km towards H2O2 was 31.82 mmol/L. The enzyme activity was strongly inhibited by sodium dodecyl sulfate(SDS), oxalic, Ag+ and Cu2+, but was activated by low concentration of Pb2+.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2015年第1期140-145,共6页
Food Science
基金
重庆市科委科技攻关重点项目(CSTC2011AB1027)
关键词
大豆
过氧化氢酶
分离纯化
性质
soybean
catalase
isolation and purification
characterization
