Troitskiĭ A V, Lysenko A M
Biokhimiia. 1975 Nov-Dec;40(6):1282-91.
Free T2 bacteriophage lysozyme is isolated and purified from 80 l portion of phagolysate by means of ballast protein and bacterial debris precipitation with rivanol, two-stage fractionation on amberlit IRC-50 and chromatography on CM-Sephadex C-50. Purified enzyme is homogenous under polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate, it has a molecular weight value similar to that in the literature. A presence of two active enzyme forms (I and II) is demonstrated. They can be separated by means of analytical electrophoresis in polyacrylamide gel at pH 4,5 and of ion-exchange chromatography on Amberlite IRC-50. T2 lysozymes I and II do not differ in their amino acid composition, ORD parameters, and they are not interconversible. Heterogeneity of phage lysozyme is shown not to be an artefact and to be due neither to heterogeneity of the initial phage poluation, nor to aggregation and to oxidation of enzyme SH-groups. The content of alpha-helix regions, as estimated by ORD is higher in phage lysozyme than in hen egg-white lysozyme, which evidences that these proteins are non-homologous.
通过用利凡诺沉淀杂质蛋白和细菌碎片、在Amberlit IRC - 50上进行两步分级分离以及在CM - Sephadex C - 50上进行色谱分离，从80升吞噬溶酶体部分中分离并纯化出游离的T2噬菌体溶菌酶。在十二烷基硫酸钠存在下，纯化后的酶在聚丙烯酰胺凝胶电泳中呈均一状态，其分子量值与文献报道的相似。证明存在两种活性酶形式（I和II）。它们可以通过在pH 4.5的聚丙烯酰胺凝胶中进行分析电泳以及在Amberlite IRC - 50上进行离子交换色谱分离。T2溶菌酶I和II在氨基酸组成、ORD参数方面没有差异，并且它们不能相互转化。噬菌体溶菌酶的异质性表明不是人为造成的，既不是由于初始噬菌体群体的异质性，也不是由于酶的SH基团的聚集和氧化。通过ORD估计，噬菌体溶菌酶中α - 螺旋区域的含量高于鸡蛋清溶菌酶，这证明这些蛋白质是非同源的。
