Bacillus amyloliquefaciens phage endolysin can enhance permeability of Pseudomonas aeruginosa outer membrane and induce cell lysis
Bacillus amyloliquefaciens phage endolysin can enhance permeability of Pseudomonas aeruginosa outer membrane and induce cell lysis
复制标题
DOI：
10.1007/s00253-003-1522-1
复制
发表时间：
2004-07-01
影响因子：
5
通讯作者：
Tanji, Y
中科院分区：
文献类型：
--
作者：
Orito, Y;Morita, M;Tanji, Y
为了确定解淀粉芽孢杆菌噬菌体内溶素 C 末端区域对铜绿假单胞菌裂解的功能，对铜绿假单胞菌外膜的通透性进行了分析。将 Glu-15 替换为 His (E15H)，将 Thr-32 替换为 Glu (T32E) 至芽孢杆菌噬菌体内溶素中。 E15H 和 T32E 取代均不会诱导酶活性和抗菌活性。 Glu-15 和 Thr-32 这两个被认为是酶的活性中心。将纯化的 E15H 和 T32E 蛋白添加到铜绿假单胞菌细胞中，诱导细胞释放周质 β-内酰胺酶，表明这两种蛋白都增强了外膜的透化作用。然而，向铜绿假单胞菌细胞添加E15H和T32E蛋白并没有诱导细胞释放细胞质ATP。这些结果表明细胞内溶素的抗菌活性需要铜绿假单胞菌外膜透化的C端增强和N端酶活性。
To determine the function of the C-terminal region of Bacillus amyloliquefaciens phage endolysin on Pseudomonas aeruginosa lysis, the permeabilization of the outer membrane of P. aeruginosa was analyzed. Glu-15 to His (E15H) and Thr-32 to Glu (T32E) substitutions were introduced into the Bacillus phage endolysin. Neither E15H nor T32E substitution induced enzymatic and antibacterial activities. These two, Glu-15 and Thr-32, were considered to be the active center of the enzyme. The addition of purified E15H and T32E proteins to P. aeruginosa cells induced the release of periplasmic beta-lactamase from the cells, indicating that both proteins enhance permeabilization of the outer membrane. However, the addition of E15H and T32E proteins to P. aeruginosa cells did not induce the release of cytoplasmic ATP from the cells. These results indicate that the antibacterial activity of the endolysin requires both the C-terminal enhancement of the permeabilization of the P. aeruginosa outer membrane and N-terminal enzymatic activity.