Sugai M, Komatsuzawa H, Miyake Y, Suginaka H
Department of Microbiology, Hiroshima University School of Dentistry, Japan.
Zentralbl Bakteriol. 1991 Jun;275(2):156-61. doi: 10.1016/s0934-8840(11)80062-9.
Bacteriolytic enzymes of different bond specificities, denatured by sodium dodecyl sulphate (SDS), were electrophoresed in polyacrylamide gels containing bacterial cells, then renatured after removal of SDS by diffusion. Enzyme activity was seen in sharp transparent bands resulting from bacteriolysis in the gels, while these sections containing bacterial cells appeared cloudy. Bacteriolytic enzymes including staphylococcal endo-beta-N-acetylglucosaminidase, lysozyme (N-acetylmuramidase), and lysostaphin (endopeptidase) were detected. The major bacteriolytic enzymes of Staphylococcus spp. were identified in gels after electrophoresis of crude enzyme preparations. This demonstrates the wide applicability of this method to the study of staphylococcal bacteriolytic enzymes. However, it should be noted that the method will fail to detect activities of bacteriolytic enzymes which are irreversibly inhibited by SDS.
具有不同键特异性的溶菌酶，经十二烷基硫酸钠（SDS）变性后，在含有细菌细胞的聚丙烯酰胺凝胶中进行电泳，然后通过扩散去除SDS后复性。在凝胶中细菌溶解产生的清晰透明条带中可见酶活性，而这些含有细菌细胞的部分则显得浑浊。检测到包括葡萄球菌内切-β-N-乙酰氨基葡萄糖苷酶、溶菌酶（N-乙酰胞壁酸酶）和溶葡萄球菌素（内肽酶）在内的溶菌酶。在粗酶制剂电泳后，在凝胶中鉴定出葡萄球菌属的主要溶菌酶。这证明了该方法在葡萄球菌溶菌酶研究中的广泛适用性。然而，应该注意的是，该方法将无法检测到被SDS不可逆抑制的溶菌酶的活性。
