摘要
从辣根块茎分离和纯化了辣根过氧化物酶（ＨＲＰ），纯化程序包括匀浆、加热、硫酸铵分级沉淀、丙酮分级沉淀和ＣＭ－２３阴离子交换层析。纯化的ＨＲＰ的Ｋ４值为２．５×１０５，ＲＺ值是３．００，酶的比活４０６Ｕ／ｍｇ蛋白。它的ＳＤＳ－ＰＡＧＥ显示一条带，酶亚基分子量为４０ＫＤ，等电聚焦电泳呈现三条带，酶活性染色呈现多条带，表明有同工酶存在。对酶的分离纯化作了一些改进，如加氨水保持ｐＨ值，加入５ｍｍｏｌ／Ｌ的硫酸铁溶液，改变洗脱缓冲液的成分等。
Peroxidase (HRP,EC 1 11 1 7) was isolated and purified from horseradish tubes.The purification steps included centrafuge,ammonium sulphate frationing,heating 20 min at 50℃,acetone fractioning and CM-23 cellous anion exchange chromatography.The purified HRP has a K 4 of 2 5×10 5 and a RZ of 3 19.Its specific activity was 406 U/mg protein.SDS-PAGE shown one band.It also revealed that HRP subunit has a molecular of 40 KD.IEF-PAGE conformed three bands,which meant there existed three isozymes at least.The HRP activity dye indicated more isozymes existed.Some improving methods were made such as controlling pH with NH 3·H 2O,adding 5 mmol/L Fe 3+ and changing elution buffer.
出处
《安徽农业大学学报》
CAS
CSCD
1998年第3期313-316,共4页
Journal of Anhui Agricultural University
关键词
辣根过氧化物酶
分离
制备
纯化
Horseradish peroxidase (HRP) Isolation Preparation Purification
