Manoharadas Salim, Witte Angela, Bläsi Udo
Max F. Perutz Laboratories, Department of Microbiology and Immunobiology, University of Vienna, Dr. Bohrgasse 9, A-1030 Vienna, Austria.
J Biotechnol. 2009 Jan 1;139(1):118-23. doi: 10.1016/j.jbiotec.2008.09.003. Epub 2008 Sep 26.
Phage lytic enzymes (enzybiotics) have gained attention as prospective tools to eradicate Gram-positive pathogens resistant to antibiotics. Attempts to purify the P16 endolysin of Staphylococcus aureus phage P68 were unsuccessful owing to the poor solubility of the protein. To overcome this limitation, we constructed a chimeric endolysin (P16-17) comprised of the inferred N-terminal d-alanyl-glycyl endopeptidase domain and the C-terminal cell wall targeting domain of the S. aureus phage P16 endolysin and the P17 minor coat protein, respectively. The domain swapping approach and the applied purification procedure resulted in soluble P16-17 protein, which exhibited antimicrobial activity towards S. aureus. In addition, P16-17 augmented the antimicrobial efficacy of the antibiotic gentamicin. This synergistic effect could be useful to reduce the effective dose of aminoglycoside antibiotics.
噬菌体裂解酶（酶抗菌剂）作为根除对抗生素耐药的革兰氏阳性病原体的潜在工具已受到关注。由于金黄色葡萄球菌噬菌体P68的P16内溶素蛋白溶解性差，纯化该蛋白的尝试未成功。为克服这一限制，我们构建了一种嵌合内溶素（P16-17），其分别由金黄色葡萄球菌噬菌体P16内溶素的推定N端d-丙氨酰-甘氨酰内肽酶结构域和C端细胞壁靶向结构域以及P17次要衣壳蛋白组成。结构域交换方法和应用的纯化程序产生了可溶性P16-17蛋白，该蛋白对金黄色葡萄球菌具有抗菌活性。此外，P16-17增强了抗生素庆大霉素的抗菌效果。这种协同效应可能有助于降低氨基糖苷类抗生素的有效剂量。
