猪凝血酶的分离纯化及部分性质研究

PURIFICATION AND CHARACTERIZATION OF THROMBIN FROM PORCINE PLASMA

经等电点沉淀,DEAE离子交换纤维素柱层析和Sephadex G-100 分子筛层析从猪血浆中纯化得到猪凝血酶,酶比活力为2 000 U/m g,纯化倍数为100,酶活力回收为56% ,酶反应的最适pH 和温度分别为7.0 和37℃,SephadexG-100层析测得酶分子量为36 000。猪凝血酶的稳定性较差,40℃保温27 h, 酶活力下降84% 。高浓度的盐对酶稳定性有较大的影响,在2 m ol/LNaCl下放置3 h,酶活力下降50% 。

A thrombin (EC 3.4.21.5) was purified from porcine plasma by precipitation at isoelectric point, ion exchange chromatography on DEAE cellulose column and followed by gel filtration on Sephadex G 100. The purified thrombin specific activity was 2 000 U/mg, and the purification fold and recovery were 100 and 56%, respectively. The M r of the porcine thrombin was 36 000 as determined by gel filtration on Sephadex G 100. The enzyme was not stable at 40℃, it lost 84% activity incubated 27 h. The thrombin lost 50% activity at 2 mol/L NaCl for 3 h.