柱状假丝酵母脂肪酶同工酶的一种高效液相色谱分离新方法  

Separation and Characterization of Candida Rugosa Lipase Isoenzymes by High Performance Liquid Chromatograpy

作　　者：胡霄雪[1] 辛嘉英[1] 徐毅[1] 崔俊儒[1] 李树本[1] 夏春谷[1] 

机构地区：[1]中国科学院兰州化学物理研究所羰基合成与选择氧化国家重点实验室,兰州730000

出　　处：《分析化学》2002年第8期981-983,共3页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金重点资助项目 ( 2 99330 4 0 )

摘　　要：建立一种“疏水界面亲和色谱”分离柱状假丝酵母脂肪酶同工酶的高效液相色谱新方法。将商品化的CRL经离子交换色谱分离为两个同工酶组分 (CRLA和CRLB) ,在极低离子强度下 ,根据同工酶活性中心周围处于“开放”构象的疏水腔具亲疏水界面的特性 ,用疏水界面亲和色谱在NucleosilC4 (10 μ ,3 0 0 ,2 5 0× 4.60mm)柱上将CRLA和CRLB都分离为 4种同工酶组分。A high performance liquid chromatographic method, 'hydrophobic interfacial affinity chromatography' to separate commercial Candida rugosa lipase has be en established. Commercial Candida rugosa lipase has been separated into two distinct fractions (CRLA and CRLB) by anion-exchange chromatography. Based on the mechanism of lipase adsorption, on, solid hydrophobic interfaces at low ionic strength. The 'hydrophobic interfacial affinity chromatography' was, employed for the fine, separation of CRL isoenzymes. At low ionic strength, both CRLA and CRLB can be separated into four, isoenzymes on the column of Nucleosil C-4. They have different hydrophobicity and UV spectra, suggest that the-open extent of large hydrophobic pocket of the four isoenzymes may be different. This method seems to be very suitable for meticulous separation of such slightly different isoenzymes fractions.

关 键 词：疏水界面亲和色谱 高效液相色谱 分离 柱状假丝酵母脂肪酶同工酶 

分 类 号：O658[理学—分析化学]