摘要
采用简单的两步法—离子交换层析和疏水层析法,对Candida sp.99-125脂肪酶进行了纯化,比活提高了10.0倍,达到27200U/mg,回收率为35.5%.SDS-PAGE电泳分析显示该酶的分子量约为38kDa.酶学性质研究表明,该酶最适反应温度为40℃,最适反应pH值为8.5,在室温下具有良好的稳定性.钙离子和Tween80能够促进提高脂肪酶的活性,而铁离子、铜离子和SDS对其有明显的抑制作用.
The extracellular lipase from Candida sp. 99-125 was purified by ion exchange chromatography and hydrophohic interaction chromatography. The results showed that the specific activity of the purified enzyme was raised by 10.0 times and the activity recovery was 35.5%. The molecular weight of the purified lipase was determined to be about 38 kDa by SDS-PAGE in Coomassie brilliant blue staining. The optimum pH and temperature of the purified enzyme were 8.5 and 40 ℃ respectively. The purified lipase remained 95% activity after incubated for 2 d at room temperature （20 ℃）. Fe^2＋, Cu^2＋ and SDS had an inhibitory effect on lipase activity, whereas Ca^2＋ salts and Tween80 increased it.
出处
《过程工程学报》
EI
CAS
CSCD
北大核心
2007年第1期141-144,共4页
The Chinese Journal of Process Engineering
基金
国家自然科学基金资助项目(编号:20576013
20325622)
国家重点基础研究发展规划(973)基金资助项目(编号:2003CB716002)
国家'十五'科技攻关基金资助项目(编号:2004BA411B05)
关键词
脂肪酶
纯化
层析
lipase
purification
chromatography
