Ochi N, Azegami M, Ishii S I
J Biochem. 1978 Mar;83(3):727-36. doi: 10.1093/oxfordjournals.jbchem.a131966.
A bacteriolytic enzyme, PR1-lysozyme, has been purified from the lysate of mitomycin C-induced pyocinogenic Pseudomonas aeruginosa, by acrinol treatment, Amberlite CG-50 chromatography, ammonium sulfate fractionation, Sephadex G-100 gel filtration and two cycles of SP-Sephadex C-50 chromatography. Homogeneity of the preparation was demonstrated by three electrophoretic techniques. PR1-lysozyme is a basic protein (pI, 9.4) and consists of a single polypeptide chain having a molecular weight of 24,000. The amino acid composition of the protein was analyzed, and no cystein residue was found among more than 210 amino acid residues. The optimum pH for enzymatic activity was 6.4 and the enzyme exhibited about 50 to 70 times greater specific activity than hen egg-white lysozyme when assayed with chloroform-killed P. aeruginosa as a substrate. By analyzing the products of enzymatic action on purified peptidoglycan of P. aeruginosa, the enzyme was identified as an N-acetylmuramidase, i.e., the same classification as hen-egg-white lysozyme. PR1-lysozyme did not show any activity towards intact cells of gram-positive and gram-negative bacteria tested. However, the enzyme was able to lyse chloroform-killed gram-negative and gram-positive bacteria.
一种溶菌酶PR1 - 溶菌酶，已从丝裂霉素C诱导产生绿脓菌素的铜绿假单胞菌的裂解物中通过以下步骤纯化得到：用依沙吖啶处理、Amberlite CG - 50柱色谱法、硫酸铵分级分离、Sephadex G - 100凝胶过滤以及两个循环的SP - Sephadex C - 50柱色谱法。通过三种电泳技术证明了该制剂的均一性。PR1 - 溶菌酶是一种碱性蛋白质（pI为9.4），由一条分子量为24,000的单一多肽链组成。分析了该蛋白质的氨基酸组成，在210多个氨基酸残基中未发现半胱氨酸残基。酶活性的最适pH为6.4，当以氯仿灭活的铜绿假单胞菌为底物进行测定时，该酶的比活性比鸡蛋清溶菌酶高约50至70倍。通过分析该酶对纯化的铜绿假单胞菌肽聚糖的作用产物，确定该酶为N - 乙酰胞壁酸酶，即与鸡蛋清溶菌酶属于同一分类。PR1 - 溶菌酶对所测试的革兰氏阳性菌和革兰氏阴性菌的完整细胞均无活性。然而，该酶能够裂解氯仿灭活的革兰氏阴性菌和革兰氏阳性菌。
