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前言：目的从延胡索(Rhizoma corydalis)中分离纯化具有溶解纤维蛋白活性的单体蛋白单一组分,并对粗酶性质做初步研究。方法利用硫酸铵分级沉淀和阴离子交换色谱从延胡索中纯化纤溶酶。结果从延胡索中纯化出两种纤溶酶,表观相对分子质量分别为33 000和59 000。粗酶用丝氨酸蛋白酶抑制剂鉴定为丝氨酸蛋白酶,ED-TA对酶活没有抑制作用。延胡索纤溶酶粗酶体外溶纤性质主要表现为激酶活性,在4~50℃活性稳定,50~75℃酶活有所下降;最适pH值为8。0,在pH 3。0左右,仍能保留大部分酶活;能抵抗胰蛋白酶水解和胃蛋白酶的作用。结论延胡索纤溶酶性质稳定,有希望研发成为一种溶栓新药。 Purpose To purify the fibrinolytic enzyme from Rhizoma corydalis(yanhusuo) and to analyze its properties.Methods Purifying the enzyme by the ammonium sulfate fractionation and DEAE-Sepharose fast flow chromatography.Results Two components of fibrinolytic enzyme(RCFE-1,RCFE-2)were isolated.The purified enzymes showed a single protein band in the SDS-PAGE.Their relative molecular weights were 33 000 and 59 000 respectively.Crude enzymes were completely inhibited by phenylmethysulfonyl fluoride(PMSF) and not b... |
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