Cell wall substrate specificity of six different lysozymes and lysozyme inhibitory activity of bacterial extracts
Cell wall substrate specificity of six different lysozymes and lysozyme inhibitory activity of bacterial extracts
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DOI：
10.1111/j.1574-6968.2006.00240.x
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发表时间：
2006-06-01
影响因子：
2.1
通讯作者：
Michiels, CW
中科院分区：
文献类型：
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作者：
Nakimbugwe, D;Masschalck, B;Michiels, CW
我们已经研究了6种不同的溶菌酶的特异性肽聚糖底物通过提取的一些革兰氏阴性菌和Micrococcus lysodeikticus与氯仿/Tris-HCl缓冲液（氯仿/缓冲液）。溶菌酶包括两种市售的（鸡蛋白色溶菌酶或HEWL，和来自球孢链霉菌的变溶菌素或M1L），和四种色谱纯化的（噬菌体λ溶菌酶或LaL，噬菌体T4溶菌酶或T4 L，鹅蛋白色溶菌酶或GEWL，和花椰菜溶菌酶或CFL）。HEWL对M. lysodeikticus比任何革兰氏阴性细胞壁，而相反的是发现LaL。此外，革兰氏阴性细胞壁对不同溶菌酶的敏感性也存在显着差异，即使是密切相关的物种，如大肠杆菌和鼠伤寒沙门氏菌。这些差异可能不是由于溶菌酶抑制剂如E.大肠杆菌的细胞壁底物，因为我们表明，氯仿提取有效地去除这种抑制剂。有趣的是，我们发现强抑制活性HEWL的氯仿/缓冲液提取物的鼠伤寒沙门氏菌，并在铜绿假单胞菌的提取物中的LaL，这表明其他溶菌酶抑制剂比常春藤存在，可能是广泛的革兰氏阴性菌。
We have investigated the specificity of six different lysozymes for peptidoglycan substrates obtained by extraction of a number of gram-negative bacteria and Micrococcus lysodeikticus with chloroform/Tris-HCl buffer (chloroform/buffer). The lysozymes included two that are commercially available (hen egg white lysozyme or HEWL, and mutanolysin from Streptomyces globisporus or M1L), and four that were chromatographically purified (bacteriophage lambda lysozyme or LaL, bacteriophage T4 lysozyme or T4L, goose egg white lysozyme or GEWL, and cauliflower lysozyme or CFL). HEWL was much more effective on M. lysodeikticus than on any of the gram-negative cell walls, while the opposite was found for LaL. Also the gram-negative cell walls showed remarkable differences in susceptibility to the different lysozymes, even for closely related species like Escherichia coli and Salmonella Typhimurium. These differences could not be due to the presence of lysozyme inhibitors such as Ivy from E. coli in the cell wall substrates because we showed that chloroform extraction effectively removed this inhibitor. Interestingly, we found strong inhibitory activity to HEWL in the chloroform/buffer extracts of Salmonella Typhimurium, and to LaL in the extracts of Pseudomonas aeruginosa, suggesting that other lysozyme inhibitors than Ivy exist and are probably widespread in gram-negative bacteria.