Kazanina G A, Petrova L Ia, Selezneva A A, Ruban E L, Volkova I M
Prikl Biokhim Mikrobiol. 1981 Jul-Aug;17(4):516-22.
By gel chromatography on Sephadex G-75 it has been demonstrated that the culture fluid of Geotrichum asteroides VKM F-144 contains two lipolytic components, one of which, lipase 1, eluates in the free volume of the column, whereas the second component, lipase 2, eluates in the volume close to that of the egg albumin yield. The paper describes a method for lipase 2 preparation consisting of precipitation of accompanying compounds at pH 4.2, depigmentation of the supernatant on the resin PAP in the Cl-form, selective sorption of lipase on the cation exchange resin CMC and further purification of the eluate gel chromatography and Sephadex G-100 rechromatography. The molecular weight of the lipase is estimated to be 45,000 and the isoelectric point to correspond to pH 4.3. pH optimum of action of the purified enzyme, areas of its pH- and thermostability have been determined.
通过在葡聚糖凝胶G - 75上进行凝胶色谱分析表明,星状地霉VKM F - 144的培养液含有两种脂解成分,其中一种脂酶1在柱的空体积中洗脱,而第二种成分脂酶2在接近卵清蛋白产物体积处洗脱。本文描述了一种制备脂酶2的方法,该方法包括在pH 4.2下沉淀伴随化合物,用Cl型树脂PAP对上清液进行脱色,在阳离子交换树脂CMC上选择性吸附脂酶,以及通过凝胶色谱和葡聚糖凝胶G - 100再色谱进一步纯化洗脱液。该脂酶的分子量估计为45,000,等电点对应于pH 4.3。已确定纯化酶的最适作用pH、其pH稳定性和热稳定性范围。